Research ArticleRedox-Reaction Based Spectrophotometric Assay ofIsoniazid in Pharmaceuticals

N Swamy K N Prashanth and K Basavaiah

Department of Studies in Chemistry University of Mysore Manasagangotri Mysore 570 006 India

Correspondence should be addressed to K Basavaiah kanakapurabasavaiahgmailcom

Received 21 January 2014 Accepted 25 February 2014 Published 22 April 2014

Academic Editors B N Barman G Drochioiu J V Garcia Mateo B Rittich and A Szemik-Hojniak

Copyright copy 2014 N Swamy et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Two spectrophotometric methods are described for the determination of isoniazid (INH) in pharmaceuticals In the first method(FCR method) INH is reacted with Folin-Ciocalteu reagent in Na

2CO3medium and the resulting blue colored chromogen

measured at 760 nm Iron(II) formed as a result of reaction between INH and iron(III) is made to react with ferricyanide and theresulting Prussian blue is measured at 760 nm basing the second method (FFC method) The conditions for better performanceare optimized Beerrsquos law is obeyed in the concentration ranges 05ndash10 and 02ndash30 120583g mLminus1 for FCR method and FFC methodsrespectively with corresponding molar absorptivity values of 112 times 104 and 455 times 104 L molminus1 cmminus1 The methods are validatedfor accuracy precision LOD LOQ robustness and ruggedness as per the current ICH guidelines The validated methods weresuccessfully applied to quantify INH in its commercial formulation with satisfactory results hence the methods are suitable forisoniazid determination in bulk drugs and pharmaceuticals

1 Introduction

Isoniazid (INH) (Figure 1) chemically known as pyridine-4-carboxylic acid hydrazide is an antitubercular drug nowwidely used together with rifampicin and streptomycin forthe chemotherapy of tuberculosis This has prompted manyinvestigators to devise methods for its determination in itspure form as well as in its tablet form

The drug is official in Indian Pharmacopoeia (IP) [1]British Pharmacopoeia (BP) [2] and United State Pharma-copoeia (USP) [3] IP and BP describe titration of the drugwith potassium bromate in presence of potassium bromideusing methyl red indicator USP describes HPLC methodusing L

1column (46mm times 25 cm) and a mobile phase

consisting of methanol water (40 60) (pH adjusted to 25with H

2SO4) with a flow rate of 15mLminminus1 and UV-

detection at 254 nm Apart from the above official methodsa number of methods based on several techniques arefound in the literature for INH and include titrimetry [4ndash7]voltammetry [8ndash13] ion selective electrode-potentiometry[14ndash19] amperometry [20 21] spectrofluorimetry [22 23]chemiluminescence spectrometry [24ndash33] high performance

liquid chromatography (HPLC) [34ndash37] gas chromatography(GC) [38ndash40] LCLC-MS [41] and capillary electrophoresis[42ndash44]

Visible spectrophotometry is by far the most widely usedtechnique for the assay of INH Methods were based on avariety of reaction schemes such as nucleophilic substitu-tion [45] condensation [46ndash49] charge-transfer and ion-pair [50] derivatization [51ndash54] diazo-coupling [55 56]oxidative coupling [57 58] complex formation [59ndash61] andredox followed by complexation [62ndash65] These methodssuffer from one or more of the disadvantages such as dras-tic experimental conditions use of organic solvent longerstanding time poor sensitivity and narrow linear rangeFolin-Ciocalteu (F-C) reagent or iron(III) and ferricyanidehave been widely used for the sensitive determination ofa wide-ranging phenolic and amine organic compounds ofpharmaceutical importance [66ndash74]

The aim of this work was to investigate the utility of F-Creagent and iron(III) and ferricyanide systems in the assayof INH The methods had sufficiently good accuracy andprecision and presented a simple and time-saving assay ofINH

Hindawi Publishing CorporationISRN Analytical ChemistryVolume 2014 Article ID 717019 11 pageshttpdxdoiorg1011552014717019

2 ISRN Analytical Chemistry

N

NO

H

NH2

Figure 1 Structure of INH

2 Experimental

21 Apparatus A Systronics model 166 digital spectropho-tometer (Ahmedabad India) with matched 1-cm quartz cellswas used for absorbance measurements

3 Materials

Chemicals used were of analytical reagent grade Distilledwater was used throughout the investigation Pharmaceuticalgrade INH certified to be 9985 pure was kindly providedby Cipla India Ltd Bangalore India and was used asreceived Isokin-300 (Pfizer Ltd Gandhinagar HyderabadIndia) tablets containing isoniazid 300mg with vitamin B

6

10mg were purchased from local commercial store

31 Reagents

311 Folin-Ciocalteu Reagent Aqueous solution of Folin-Ciocalteu reagent (1 1 vv) was prepared by mixing 50mL ofreagent (Merck Mumbai India) with 50mL water

312 Sodium Carbonate A 20 solution of sodium carbon-ate was prepared by dissolving 20 g compound (SD FineChem Ltd Mumbai India) in 100mL water

313 Potassium Ferricyanide A 500 120583gmLminus1 potassium fer-ricyanide solution was prepared by dissolving 50mg reagent(Glaxo Laboratory Mumbai India) in 100mL water

314 Ferric Chloride (FeCl3) A solution of 05M

FeCl3sdot6H2O was prepared by dissolving 14 g of the chemical

(SD Fine Chem Ltd Mumbai India) in 100mL of 01MHCl (Merck Mumbai India)

315 Standard Drug Solution A stock standard solution ofINH (100120583gmLminus1) was prepared by dissolving 10mg of pureINH in 100mL water appropriately diluted to 20120583gmLminus1INH for method A and 10 120583gmLminus1 INH for method B withwater and used in the assay

4 Assay Procedure

41 Method A (FCR Method) Into a series of 10mL vol-umetric flasks different aliquots of working standard INHsolution (20120583gmLminus1) ranging from 025 to 50mL equivalentto 05ndash100 120583gmLminus1 were transferred and the total volumewas brought to 5mL with water To each flask 2mL of1 1 F-C reagent and 2mL of 20 Na

2CO3solution were

successively added by means of a microburette The flaskswere stoppered and the contents were mixed well and keptat room temperature for 10min The volume was made upto the mark with water and the absorbance of each solutionwas measured at 760 nm against a reagent blank similarlyprepared in the absence of INH

42 Method B (FFC Method) Different aliquots of standardINH solution (10 120583gmLminus1) ranging from 02 to 30mL weretransferred into a series of 10mL of calibrated flasks andthe total volume was brought to 3mL with water Then15mL of 500120583gmLminus1 potassium ferricyanide and 10mLFeCl3solution were accurately added The volume was made

up to the mark with water content mixed and the flaskswere kept at room temperature After 10min the absorbancewas measured at 760 nm against reagent blank preparedsimultaneously without adding INH

Standard graph was prepared by plotting the absorbanceversus INH concentration and the concentration of theunknown was computed from the respective regressionequation derived using the absorbance-concentration data

43 Procedure for Pharmaceutical Tablets Twenty tabletswere weighed and ground into a fine powder An accuratelyweighed quantity containing 10mg of INHwas transferred toa 100mL volumetric flask 60mL water added shaken wellfor 20 minutes and made up to mark with water and thenfilteredThis solution was diluted to a 20 and 10 120583gmLminus1 INHwith water and analyzed by the recommended procedures

44 Procedure for Placebo Blank and Synthetic MixtureAnalyses A placebo blank containing starch (10mg) acacia(15mg) hydroxyl cellulose (10mg) sodium citrate (10mg)talc (20mg) magnesium stearate (15mg) and sodium algi-nate (10mg) was prepared by mixing all components intoa homogeneous mixture 10mg of the placebo blank wasaccurately weighed and its solutionwas prepared as describedunder Procedure for Pharmaceutical Tablets and then sub-jected to analysis by following the general procedure

An accurately weighed 10mg of INH was added to 10mgof placebo blank and homogenized Synthetic mixture wasquantitatively transferred into a 100mL volumetric flask andthe extract was prepared as described under the Procedurefor Pharmaceutical Tablets The resulting extract was dilutedto get 20 and 10 120583gmLminus1 INH solutions respectively Suitablealiquot of the solution was analyzed at three levels byfollowing the general assay procedure

ISRN Analytical Chemistry 3

Abso

rban

ce

Wavelength (nm)

0

01

02

03

04

05

A

B

500 550 600 650 700 750 800 850 900

(a)

Abso

rban

ce

Wavelength (nm)

0

01

02

03

04

05

650 670 690 710 730 750 770 790 810

(b)

Figure 2 (a) Absorption spectra of A reaction product of INH (6 120583gmLminus1) with F-C reagent inNa2CO3solution andB blank (b) Absorption

spectra for method B reaction product of INH (125 120583gmLminus1) with ferric chloride and ferricyanide solution (-998771-) and blank (-Icircsect-)

5 Validation of Method

The method was validated according to the proceduresdescribed in ICH guidelines [75] for the validation of ana-lytical methodsLimits of Detection (LOD) and Quantification (LOQ) Thelimits of detection (LOD) and quantification (LOQ)were cal-culated according to the ICH guidelines using the formulae

LOD = 33119878119887 119871119874119876 = 10

119878

119887 (1)

where 119878 is the standard deviation of blank absorbance valuesand 119887 is the slope of the calibration plot

51 Precision and Accuracy The intraday precision was eval-uated by analyzing INH solution at three different levels Sim-ilarly the interday precision was evaluated on five consecutivedays (119899 = 5) In each case and for each concentration meanvalue of the INH found and the relative standard deviations(RSD) were calculated The accuracy of the methods wasdetermined by the percent mean deviation between theobtained and known concentration of INH

52 Robustness and Ruggedness Method robustness wasstudied by making small changes in the optimized experi-mental variables and their effect on the absorbance was eval-uated by calculating the percentage RSD values In order todetermine the method ruggedness analyses were performedusing three instruments and also by three analysts with thesame instrument

6 Results and Discussion

The proposed FCR method is based on the formation of ablue colored chromogen following the reduction of phos-phomolybdotungstic mixed acid of the F-C reagent [73] byINH in the presence of sodium carbonate which could bemeasured at 760 nm The mixed acids in the F-C reagent

Table 1 Sensitivity and regression parameters

Parameter Method A Method B120582max nm 760 760Color stability min 60 90Linear range 120583gmLminus1 05ndash100 02ndash30Molar absorptivity (120576) Lmolminus1 cmminus1 112 times 104 455 times 104

Sandell sensitivitylowast 120583g cmminus2 00123 0003Limit of detection (LOD) 120583gmLminus1 003 014Limit of quantification (LOQ) 120583gmLminus1 009 042Regression equation 119884lowastlowast

Intercept (119886) 00131 00198Slope (119887) 00905 03176Standard deviation of 119886 (119878

119886) 998 times 10minus2 998 times 10minus2

Standard deviation of 119887 (119878119887) 124 times 10minus2 398 times 10minus2

Regression coefficient (119903) 09971 09979lowastLimit of determination as the weight in 120583gmLminus1 of solution whichcorresponds to an absorbance of A = 0001 measured in a cuvette of cross-sectional area 1 cm2 and 119897 = 1 cmlowastlowast119884 = 119886 + 119887119883 where 119884 is the absorbance119883 is concentration in 120583gmLminus1 119886

is intercept and 119887 is slope

are the final chromogen and involve the following chemicalspecies

3H2O sdot P2O5sdot 13WO

3sdot 5MoO

3sdot 10H2O

3H2O sdot P2O5sdot 14WO

3sdot 4MoO

3sdot 10H2O

(2)

INH probably affects reduction of oxygen atoms fromtungstate andor molybdate in the F-C reagent by producingone or more possible reduced species which have character-istic intense blue color

The FFCmethod involves the redox reaction of INHwithferric chloride in the presence of potassium ferricyanideunder mild acidic conditions to produce a blue colour withmaximum absorption at 760 nm The first step in the colourdevelopment is the reduction of iron(III) of ferric chloride toiron(II) which subsequently reacts with ferricyanide to formPrussian blue

4 ISRN Analytical ChemistryAb

sorb

ance

1 1 FC-reagent (mL)

03

04

05

06

0 05 10 15 20 25 30

(a)

0

02

04

06

0 1 2 3 4 5

Abso

rban

ce

15 Na2CO3 (mL)

(b)

Abso

rban

ce

02

03

04

05

0 10 20 30 40 50 60

Time (min)

(c)

Figure 3 (a) Effect of volumes of F-C reagent (1 1) on the reactionproduct with INH (6 120583gmLminus1) in Na

2CO3solution for method A

(b) Effect of 20 Na2CO3on color formation (6120583gmLminus1 INH) for

method A (c) Reaction time and stability of colored species formethod A (6120583gmLminus1 INH)

7 Method Development

71 Spectral Characteristics The intensely blue colored prod-uct (molybdenum-tungsten mixed acid blue in FCR methodand Prussian blue in FFC method) formed in both methodsexhibited maximum absorption at 760 nm The absorptionspectra of the blue colored products and of the reagent blanksare shown in Figures 2(a) and 2(b)

Abso

rban

ce

035

040

045

050

FeCl3 (mL)05 10 15 20 25

(a)

Abso

rban

ce

030

035

040

045

050

Ferricyanide (mL)0 10 15 20 25

(b)

Abso

rban

ce

020

040

030

050

Reaction time (min)0 15 30 45 60 75 90 105 120

(c)

Figure 4 (a) Effect of volume of FeCl3for method B (15 120583gmLminus1

INH) (b) Effect of volume of ferricyanide formethod B (15120583gmLminus1INH) (c) Reaction time and stability of colored species for methodB (15 120583gmLminus1 INH)

72 Optimization of Experimental Variables A series ofpreliminary experiments necessary for rapid and quantitativeformation of colored products to achieve the maximumstability and sensitivity were performed Optimum conditionwas achieved by varying one parameter at a time whilekeeping other parameters constant and observing its effect onthe absorbance at 760 nm

ISRN Analytical Chemistry 5

Table 2 Evaluation of intraday and interday accuracy and precision

MethodINH Taken (120583gmLminus1) Intraday accuracy and precision (119899 = 7) Interday accuracy and precision (119899 = 5)

INH Founda (120583gmLminus1) RSDb

REc

INH found (120583gmLminus1) RSDb

REc

A30 296 124 089 307 234 21260 611 184 103 612 200 25390 914 156 152 915 167 200

B10 101 102 192 103 300 18515 147 201 115 154 267 12720 204 192 147 205 250 162

aMean value of 7 determinations brelative standard deviation () crelative error ()

Table 3 Method robustness and ruggedness expressed as intermediate precision

Method Nominal concentrationRobustness Ruggedness

Reaction timeslowast(119899 = 3)

Volumes of FCreagent

Volumesof ferricyanide

Interanalysts(119899 = 3)

Interinstruments(119899 = 3)

A30 145 169 mdash 220 23260 172 135 mdash 174 19290 210 171 mdash 244 257

B10 172 mdash 159 178 21215 184 mdash 161 211 16220 212 mdash 172 107 254

lowastReaction times were 10 plusmn 2min for both methods Volumes of FC regent added were 2 plusmn 02mL for method A volumes of ferricyanide added were 15 plusmn01mL for method B

73 Optimization for FCR Method

731 Effect of Concentration of F-C Reagent Several experi-ments were carried out to study the influence of F-C reagentconcentration on the color development and the obtainedresults are shown in Figure 3(a) It is apparent that 10 to50mL of reagent gave the maximum color intensity thus30mL of reagent was used throughout the investigation

732 Selection of Reaction Medium and Optimization of theBase To select a suitable medium for the reaction differentaqueous bases such as sodium hydroxide sodium carbon-ate or bicarbonate sodium acetate and sodium hydrogenphosphate were investigated Better results were obtainedwith sodium carbonate In order to determine the optimumconcentration of Na

2CO3 different volumes of 20 Na

2CO3

solution (0ndash5mL)were attempted at a constant concentrationof INH (6 120583gmLminus1) and the results of the observation areshown in Figure 3 It was found that different volumes rang-ing from 10 to 30mL of 20 Na

2CO3were optimum thus

20mL was used throughout the work (given in Figure 3(b))Maximum color development was obtained in 10min

after mixing the reactants and the color was stable for atleast 60min thereafter (Figure 3(c)) The sequence of orderof addition of the reactants had significant effect on theabsorbance value So the order used in the general procedureshould be followed for maximum absorbance

74 Optimization for FFC Method To optimize the concen-trations of ferricyanide and ferric chloride reagents differentvolumes of these reagents were used with a fixed concentra-tion of INH Constant absorbance was found with 10mL of05MFeCl

3and 15mLof 500120583gmLminus1 ferricyanide solutions

as shown in Figures 4(a) and 4(b)Colour development was complete in 10min and stable

for the next 90min (Figure 4(c)) Different results wereobtained when different order of addition of reactants wasfollowed The order of addition of reactants followed in therecommended procedure resulted in rapid color formationwith maximum sensitivity and stability

8 Method Validation

81 Linearity Sensitivity Limits of Detection and Quantifi-cation A linear correlation was found between absorbanceat 120582max and concentration of INH in the ranges given inTable 1 Regression analysis of the Beerrsquos law data using themethod of least squares was made to evaluate the slope (119887)intercept (119886) and correlation coefficient (119903) for each systemand the analytical results obtained from these investigationsare presented in Table 1 The optical characteristics such asBeerrsquos law limits molar absorptivity and Sandell sensitivityvalues are also given in Table 1 The high values of 120576 andlow values of Sandell sensitivity and LOD indicate the highsensitivity of the proposed methods

6 ISRN Analytical Chemistry

Table 4 Results of analysis of tablets by the proposed methods

Tablets analyzed Label claim (mgtablet)Foundlowast (percent of label claim plusmn SD)

Official method Proposed methodsMethod A Method B

Iskin-300 300 9986 plusmn 136

1015 plusmn 116 1012 plusmn 092

119905 = 206 119905 = 183

119865 = 137 119865 = 219

lowastMean value of five determinationsTabulated 119905-value at the 95 confidence level is 277Tabulated 119865-value at the 95 confidence level is 639

Table 5 Results of recovery study via standard addition method with tablet

Method Tablets studied INH in tablet 120583gmLminus1 Pure INH Added 120583gmLminus1 Total Found 120583gmLminus1 Pure INH recoveredlowastpercent plusmn SD

A Isokin-300296 15 441 9928 plusmn 203

296 30 601 1009 plusmn 312

296 45 756 1013 plusmn 299

B Isokin-300101 05 151 9985 plusmn 225

101 10 204 1013 plusmn 232

101 15 255 1017 plusmn 314

lowastMean value of three determinations

82 Intraday and Interday Precision and Accuracy The pre-cision and accuracy of the proposed method were studiedby repeating the experiment seven times within the dayto determine the repeatability (intraday precision) and fivetimes on different days to determine the intermediate preci-sion (interday precision) The assay was performed for threelevels of analyte in this method The results of this studyare summarized in Table 2 The percentage relative standarddeviation (RSD) values were le201 (intraday) and le300(interday) indicating good precision of themethod Accuracywas evaluated as percentage relative error (RE) between themeasured mean concentrations and taken concentrations ofINH and it was le253 demonstrating the high accuracy ofthe proposed method

83 Selectivity A selective study was performed to determinethe effect of matrix on the absorbance by analyzing theplacebo blank In the analysis of placebo blank solution theabsorbance in each case was equal to the absorbance ofblank which revealed no interference To assess the role ofthe inactive ingredients on the assay of INH the generalprocedure was applied on the synthetic mixture extract bytaking three different concentrations of INHwithin the rangeThe percentage recovery values were in the range 963ndash1023 and 954ndash1018 with RSD lt4 indicating clearly thenoninterference from the inactive ingredients in the assay ofINH

84 Robustness and Ruggedness The robustness of themethod was evaluated by making small incremental changesin the volume of reagent and reaction time and the effectsof the changes were studied by measuring the absorbance ofthe colored product The changes had negligible influence

on the results as revealed by small intermediate precisionvalues expressed as RSD (le254) Method ruggednesswas demonstrated having the analysis done by three analystsand also by a single analyst performing analysis on threedifferent instruments in the same laboratory Intermediateprecision values (RSD) in both instances indicated accept-able ruggedness These results are presented in Table 3

85 Application to Tablets The proposed methods wereapplied to the quantification of INH in commercial tabletsThe tablets were assayed by the official BP method [2] whichdescribes titration of the drug with potassium bromate inpresence of potassium bromide using methyl red indicatorThe results obtained by the proposed methods agree wellwith the claim and also are in agreement with those of theofficial method Statistical analysis of the results did notdetect any significant difference between the performance ofthe proposed method and reference method with respect toaccuracy and precision as revealed by the Studentrsquos 119905-valueand variance ratio 119865-value The results of assay are given inTable 4

86 Recovery Study To further assess the accuracy of themethod recovery experiment was performed by applying thestandard-addition technique The recovery was assessed bydetermining the agreement between the measured standardconcentration and added known concentration to the sampleThe test was done by spiking the preanalyzed tablet INHwithpure INH at three different levels (50 100 and 150) of thecontent present in the preparation and the total was found bythe proposedmethod Each test was repeated three timesTherecovery percentage values ranged between 9928 and 1017with standard deviation in the range 099ndash132 Closeness

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

2 ISRN Analytical Chemistry

N

NO

H

NH2

Figure 1 Structure of INH

2 Experimental

21 Apparatus A Systronics model 166 digital spectropho-tometer (Ahmedabad India) with matched 1-cm quartz cellswas used for absorbance measurements

3 Materials

Chemicals used were of analytical reagent grade Distilledwater was used throughout the investigation Pharmaceuticalgrade INH certified to be 9985 pure was kindly providedby Cipla India Ltd Bangalore India and was used asreceived Isokin-300 (Pfizer Ltd Gandhinagar HyderabadIndia) tablets containing isoniazid 300mg with vitamin B

6

10mg were purchased from local commercial store

31 Reagents

311 Folin-Ciocalteu Reagent Aqueous solution of Folin-Ciocalteu reagent (1 1 vv) was prepared by mixing 50mL ofreagent (Merck Mumbai India) with 50mL water

312 Sodium Carbonate A 20 solution of sodium carbon-ate was prepared by dissolving 20 g compound (SD FineChem Ltd Mumbai India) in 100mL water

313 Potassium Ferricyanide A 500 120583gmLminus1 potassium fer-ricyanide solution was prepared by dissolving 50mg reagent(Glaxo Laboratory Mumbai India) in 100mL water

314 Ferric Chloride (FeCl3) A solution of 05M

FeCl3sdot6H2O was prepared by dissolving 14 g of the chemical

(SD Fine Chem Ltd Mumbai India) in 100mL of 01MHCl (Merck Mumbai India)

315 Standard Drug Solution A stock standard solution ofINH (100120583gmLminus1) was prepared by dissolving 10mg of pureINH in 100mL water appropriately diluted to 20120583gmLminus1INH for method A and 10 120583gmLminus1 INH for method B withwater and used in the assay

4 Assay Procedure

41 Method A (FCR Method) Into a series of 10mL vol-umetric flasks different aliquots of working standard INHsolution (20120583gmLminus1) ranging from 025 to 50mL equivalentto 05ndash100 120583gmLminus1 were transferred and the total volumewas brought to 5mL with water To each flask 2mL of1 1 F-C reagent and 2mL of 20 Na

2CO3solution were

successively added by means of a microburette The flaskswere stoppered and the contents were mixed well and keptat room temperature for 10min The volume was made upto the mark with water and the absorbance of each solutionwas measured at 760 nm against a reagent blank similarlyprepared in the absence of INH

42 Method B (FFC Method) Different aliquots of standardINH solution (10 120583gmLminus1) ranging from 02 to 30mL weretransferred into a series of 10mL of calibrated flasks andthe total volume was brought to 3mL with water Then15mL of 500120583gmLminus1 potassium ferricyanide and 10mLFeCl3solution were accurately added The volume was made

up to the mark with water content mixed and the flaskswere kept at room temperature After 10min the absorbancewas measured at 760 nm against reagent blank preparedsimultaneously without adding INH

Standard graph was prepared by plotting the absorbanceversus INH concentration and the concentration of theunknown was computed from the respective regressionequation derived using the absorbance-concentration data

43 Procedure for Pharmaceutical Tablets Twenty tabletswere weighed and ground into a fine powder An accuratelyweighed quantity containing 10mg of INHwas transferred toa 100mL volumetric flask 60mL water added shaken wellfor 20 minutes and made up to mark with water and thenfilteredThis solution was diluted to a 20 and 10 120583gmLminus1 INHwith water and analyzed by the recommended procedures

44 Procedure for Placebo Blank and Synthetic MixtureAnalyses A placebo blank containing starch (10mg) acacia(15mg) hydroxyl cellulose (10mg) sodium citrate (10mg)talc (20mg) magnesium stearate (15mg) and sodium algi-nate (10mg) was prepared by mixing all components intoa homogeneous mixture 10mg of the placebo blank wasaccurately weighed and its solutionwas prepared as describedunder Procedure for Pharmaceutical Tablets and then sub-jected to analysis by following the general procedure

An accurately weighed 10mg of INH was added to 10mgof placebo blank and homogenized Synthetic mixture wasquantitatively transferred into a 100mL volumetric flask andthe extract was prepared as described under the Procedurefor Pharmaceutical Tablets The resulting extract was dilutedto get 20 and 10 120583gmLminus1 INH solutions respectively Suitablealiquot of the solution was analyzed at three levels byfollowing the general assay procedure

ISRN Analytical Chemistry 3

Abso

rban

ce

Wavelength (nm)

0

01

02

03

04

05

A

B

500 550 600 650 700 750 800 850 900

(a)

Abso

rban

ce

Wavelength (nm)

0

01

02

03

04

05

650 670 690 710 730 750 770 790 810

(b)

Figure 2 (a) Absorption spectra of A reaction product of INH (6 120583gmLminus1) with F-C reagent inNa2CO3solution andB blank (b) Absorption

spectra for method B reaction product of INH (125 120583gmLminus1) with ferric chloride and ferricyanide solution (-998771-) and blank (-Icircsect-)

5 Validation of Method

The method was validated according to the proceduresdescribed in ICH guidelines [75] for the validation of ana-lytical methodsLimits of Detection (LOD) and Quantification (LOQ) Thelimits of detection (LOD) and quantification (LOQ)were cal-culated according to the ICH guidelines using the formulae

LOD = 33119878119887 119871119874119876 = 10

119878

119887 (1)

where 119878 is the standard deviation of blank absorbance valuesand 119887 is the slope of the calibration plot

51 Precision and Accuracy The intraday precision was eval-uated by analyzing INH solution at three different levels Sim-ilarly the interday precision was evaluated on five consecutivedays (119899 = 5) In each case and for each concentration meanvalue of the INH found and the relative standard deviations(RSD) were calculated The accuracy of the methods wasdetermined by the percent mean deviation between theobtained and known concentration of INH

52 Robustness and Ruggedness Method robustness wasstudied by making small changes in the optimized experi-mental variables and their effect on the absorbance was eval-uated by calculating the percentage RSD values In order todetermine the method ruggedness analyses were performedusing three instruments and also by three analysts with thesame instrument

6 Results and Discussion

The proposed FCR method is based on the formation of ablue colored chromogen following the reduction of phos-phomolybdotungstic mixed acid of the F-C reagent [73] byINH in the presence of sodium carbonate which could bemeasured at 760 nm The mixed acids in the F-C reagent

Table 1 Sensitivity and regression parameters

Parameter Method A Method B120582max nm 760 760Color stability min 60 90Linear range 120583gmLminus1 05ndash100 02ndash30Molar absorptivity (120576) Lmolminus1 cmminus1 112 times 104 455 times 104

Sandell sensitivitylowast 120583g cmminus2 00123 0003Limit of detection (LOD) 120583gmLminus1 003 014Limit of quantification (LOQ) 120583gmLminus1 009 042Regression equation 119884lowastlowast

Intercept (119886) 00131 00198Slope (119887) 00905 03176Standard deviation of 119886 (119878

119886) 998 times 10minus2 998 times 10minus2

Standard deviation of 119887 (119878119887) 124 times 10minus2 398 times 10minus2

Regression coefficient (119903) 09971 09979lowastLimit of determination as the weight in 120583gmLminus1 of solution whichcorresponds to an absorbance of A = 0001 measured in a cuvette of cross-sectional area 1 cm2 and 119897 = 1 cmlowastlowast119884 = 119886 + 119887119883 where 119884 is the absorbance119883 is concentration in 120583gmLminus1 119886

is intercept and 119887 is slope

are the final chromogen and involve the following chemicalspecies

3H2O sdot P2O5sdot 13WO

3sdot 5MoO

3sdot 10H2O

3H2O sdot P2O5sdot 14WO

3sdot 4MoO

3sdot 10H2O

(2)

INH probably affects reduction of oxygen atoms fromtungstate andor molybdate in the F-C reagent by producingone or more possible reduced species which have character-istic intense blue color

The FFCmethod involves the redox reaction of INHwithferric chloride in the presence of potassium ferricyanideunder mild acidic conditions to produce a blue colour withmaximum absorption at 760 nm The first step in the colourdevelopment is the reduction of iron(III) of ferric chloride toiron(II) which subsequently reacts with ferricyanide to formPrussian blue

4 ISRN Analytical ChemistryAb

sorb

ance

1 1 FC-reagent (mL)

03

04

05

06

0 05 10 15 20 25 30

(a)

0

02

04

06

0 1 2 3 4 5

Abso

rban

ce

15 Na2CO3 (mL)

(b)

Abso

rban

ce

02

03

04

05

0 10 20 30 40 50 60

Time (min)

(c)

Figure 3 (a) Effect of volumes of F-C reagent (1 1) on the reactionproduct with INH (6 120583gmLminus1) in Na

2CO3solution for method A

(b) Effect of 20 Na2CO3on color formation (6120583gmLminus1 INH) for

method A (c) Reaction time and stability of colored species formethod A (6120583gmLminus1 INH)

7 Method Development

71 Spectral Characteristics The intensely blue colored prod-uct (molybdenum-tungsten mixed acid blue in FCR methodand Prussian blue in FFC method) formed in both methodsexhibited maximum absorption at 760 nm The absorptionspectra of the blue colored products and of the reagent blanksare shown in Figures 2(a) and 2(b)

Abso

rban

ce

035

040

045

050

FeCl3 (mL)05 10 15 20 25

(a)

Abso

rban

ce

030

035

040

045

050

Ferricyanide (mL)0 10 15 20 25

(b)

Abso

rban

ce

020

040

030

050

Reaction time (min)0 15 30 45 60 75 90 105 120

(c)

Figure 4 (a) Effect of volume of FeCl3for method B (15 120583gmLminus1

INH) (b) Effect of volume of ferricyanide formethod B (15120583gmLminus1INH) (c) Reaction time and stability of colored species for methodB (15 120583gmLminus1 INH)

72 Optimization of Experimental Variables A series ofpreliminary experiments necessary for rapid and quantitativeformation of colored products to achieve the maximumstability and sensitivity were performed Optimum conditionwas achieved by varying one parameter at a time whilekeeping other parameters constant and observing its effect onthe absorbance at 760 nm

ISRN Analytical Chemistry 5

Table 2 Evaluation of intraday and interday accuracy and precision

MethodINH Taken (120583gmLminus1) Intraday accuracy and precision (119899 = 7) Interday accuracy and precision (119899 = 5)

INH Founda (120583gmLminus1) RSDb

REc

INH found (120583gmLminus1) RSDb

REc

A30 296 124 089 307 234 21260 611 184 103 612 200 25390 914 156 152 915 167 200

B10 101 102 192 103 300 18515 147 201 115 154 267 12720 204 192 147 205 250 162

aMean value of 7 determinations brelative standard deviation () crelative error ()

Table 3 Method robustness and ruggedness expressed as intermediate precision

Method Nominal concentrationRobustness Ruggedness

Reaction timeslowast(119899 = 3)

Volumes of FCreagent

Volumesof ferricyanide

Interanalysts(119899 = 3)

Interinstruments(119899 = 3)

A30 145 169 mdash 220 23260 172 135 mdash 174 19290 210 171 mdash 244 257

B10 172 mdash 159 178 21215 184 mdash 161 211 16220 212 mdash 172 107 254

lowastReaction times were 10 plusmn 2min for both methods Volumes of FC regent added were 2 plusmn 02mL for method A volumes of ferricyanide added were 15 plusmn01mL for method B

73 Optimization for FCR Method

731 Effect of Concentration of F-C Reagent Several experi-ments were carried out to study the influence of F-C reagentconcentration on the color development and the obtainedresults are shown in Figure 3(a) It is apparent that 10 to50mL of reagent gave the maximum color intensity thus30mL of reagent was used throughout the investigation

732 Selection of Reaction Medium and Optimization of theBase To select a suitable medium for the reaction differentaqueous bases such as sodium hydroxide sodium carbon-ate or bicarbonate sodium acetate and sodium hydrogenphosphate were investigated Better results were obtainedwith sodium carbonate In order to determine the optimumconcentration of Na

2CO3 different volumes of 20 Na

2CO3

solution (0ndash5mL)were attempted at a constant concentrationof INH (6 120583gmLminus1) and the results of the observation areshown in Figure 3 It was found that different volumes rang-ing from 10 to 30mL of 20 Na

2CO3were optimum thus

20mL was used throughout the work (given in Figure 3(b))Maximum color development was obtained in 10min

after mixing the reactants and the color was stable for atleast 60min thereafter (Figure 3(c)) The sequence of orderof addition of the reactants had significant effect on theabsorbance value So the order used in the general procedureshould be followed for maximum absorbance

74 Optimization for FFC Method To optimize the concen-trations of ferricyanide and ferric chloride reagents differentvolumes of these reagents were used with a fixed concentra-tion of INH Constant absorbance was found with 10mL of05MFeCl

3and 15mLof 500120583gmLminus1 ferricyanide solutions

as shown in Figures 4(a) and 4(b)Colour development was complete in 10min and stable

for the next 90min (Figure 4(c)) Different results wereobtained when different order of addition of reactants wasfollowed The order of addition of reactants followed in therecommended procedure resulted in rapid color formationwith maximum sensitivity and stability

8 Method Validation

81 Linearity Sensitivity Limits of Detection and Quantifi-cation A linear correlation was found between absorbanceat 120582max and concentration of INH in the ranges given inTable 1 Regression analysis of the Beerrsquos law data using themethod of least squares was made to evaluate the slope (119887)intercept (119886) and correlation coefficient (119903) for each systemand the analytical results obtained from these investigationsare presented in Table 1 The optical characteristics such asBeerrsquos law limits molar absorptivity and Sandell sensitivityvalues are also given in Table 1 The high values of 120576 andlow values of Sandell sensitivity and LOD indicate the highsensitivity of the proposed methods

6 ISRN Analytical Chemistry

Table 4 Results of analysis of tablets by the proposed methods

Tablets analyzed Label claim (mgtablet)Foundlowast (percent of label claim plusmn SD)

Official method Proposed methodsMethod A Method B

Iskin-300 300 9986 plusmn 136

1015 plusmn 116 1012 plusmn 092

119905 = 206 119905 = 183

119865 = 137 119865 = 219

lowastMean value of five determinationsTabulated 119905-value at the 95 confidence level is 277Tabulated 119865-value at the 95 confidence level is 639

Table 5 Results of recovery study via standard addition method with tablet

Method Tablets studied INH in tablet 120583gmLminus1 Pure INH Added 120583gmLminus1 Total Found 120583gmLminus1 Pure INH recoveredlowastpercent plusmn SD

A Isokin-300296 15 441 9928 plusmn 203

296 30 601 1009 plusmn 312

296 45 756 1013 plusmn 299

B Isokin-300101 05 151 9985 plusmn 225

101 10 204 1013 plusmn 232

101 15 255 1017 plusmn 314

lowastMean value of three determinations

82 Intraday and Interday Precision and Accuracy The pre-cision and accuracy of the proposed method were studiedby repeating the experiment seven times within the dayto determine the repeatability (intraday precision) and fivetimes on different days to determine the intermediate preci-sion (interday precision) The assay was performed for threelevels of analyte in this method The results of this studyare summarized in Table 2 The percentage relative standarddeviation (RSD) values were le201 (intraday) and le300(interday) indicating good precision of themethod Accuracywas evaluated as percentage relative error (RE) between themeasured mean concentrations and taken concentrations ofINH and it was le253 demonstrating the high accuracy ofthe proposed method

83 Selectivity A selective study was performed to determinethe effect of matrix on the absorbance by analyzing theplacebo blank In the analysis of placebo blank solution theabsorbance in each case was equal to the absorbance ofblank which revealed no interference To assess the role ofthe inactive ingredients on the assay of INH the generalprocedure was applied on the synthetic mixture extract bytaking three different concentrations of INHwithin the rangeThe percentage recovery values were in the range 963ndash1023 and 954ndash1018 with RSD lt4 indicating clearly thenoninterference from the inactive ingredients in the assay ofINH

84 Robustness and Ruggedness The robustness of themethod was evaluated by making small incremental changesin the volume of reagent and reaction time and the effectsof the changes were studied by measuring the absorbance ofthe colored product The changes had negligible influence

on the results as revealed by small intermediate precisionvalues expressed as RSD (le254) Method ruggednesswas demonstrated having the analysis done by three analystsand also by a single analyst performing analysis on threedifferent instruments in the same laboratory Intermediateprecision values (RSD) in both instances indicated accept-able ruggedness These results are presented in Table 3

85 Application to Tablets The proposed methods wereapplied to the quantification of INH in commercial tabletsThe tablets were assayed by the official BP method [2] whichdescribes titration of the drug with potassium bromate inpresence of potassium bromide using methyl red indicatorThe results obtained by the proposed methods agree wellwith the claim and also are in agreement with those of theofficial method Statistical analysis of the results did notdetect any significant difference between the performance ofthe proposed method and reference method with respect toaccuracy and precision as revealed by the Studentrsquos 119905-valueand variance ratio 119865-value The results of assay are given inTable 4

86 Recovery Study To further assess the accuracy of themethod recovery experiment was performed by applying thestandard-addition technique The recovery was assessed bydetermining the agreement between the measured standardconcentration and added known concentration to the sampleThe test was done by spiking the preanalyzed tablet INHwithpure INH at three different levels (50 100 and 150) of thecontent present in the preparation and the total was found bythe proposedmethod Each test was repeated three timesTherecovery percentage values ranged between 9928 and 1017with standard deviation in the range 099ndash132 Closeness

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

ISRN Analytical Chemistry 3

Abso

rban

ce

Wavelength (nm)

0

01

02

03

04

05

A

B

500 550 600 650 700 750 800 850 900

(a)

Abso

rban

ce

Wavelength (nm)

0

01

02

03

04

05

650 670 690 710 730 750 770 790 810

(b)

Figure 2 (a) Absorption spectra of A reaction product of INH (6 120583gmLminus1) with F-C reagent inNa2CO3solution andB blank (b) Absorption

spectra for method B reaction product of INH (125 120583gmLminus1) with ferric chloride and ferricyanide solution (-998771-) and blank (-Icircsect-)

5 Validation of Method

The method was validated according to the proceduresdescribed in ICH guidelines [75] for the validation of ana-lytical methodsLimits of Detection (LOD) and Quantification (LOQ) Thelimits of detection (LOD) and quantification (LOQ)were cal-culated according to the ICH guidelines using the formulae

LOD = 33119878119887 119871119874119876 = 10

119878

119887 (1)

where 119878 is the standard deviation of blank absorbance valuesand 119887 is the slope of the calibration plot

51 Precision and Accuracy The intraday precision was eval-uated by analyzing INH solution at three different levels Sim-ilarly the interday precision was evaluated on five consecutivedays (119899 = 5) In each case and for each concentration meanvalue of the INH found and the relative standard deviations(RSD) were calculated The accuracy of the methods wasdetermined by the percent mean deviation between theobtained and known concentration of INH

52 Robustness and Ruggedness Method robustness wasstudied by making small changes in the optimized experi-mental variables and their effect on the absorbance was eval-uated by calculating the percentage RSD values In order todetermine the method ruggedness analyses were performedusing three instruments and also by three analysts with thesame instrument

6 Results and Discussion

The proposed FCR method is based on the formation of ablue colored chromogen following the reduction of phos-phomolybdotungstic mixed acid of the F-C reagent [73] byINH in the presence of sodium carbonate which could bemeasured at 760 nm The mixed acids in the F-C reagent

Table 1 Sensitivity and regression parameters

Parameter Method A Method B120582max nm 760 760Color stability min 60 90Linear range 120583gmLminus1 05ndash100 02ndash30Molar absorptivity (120576) Lmolminus1 cmminus1 112 times 104 455 times 104

Sandell sensitivitylowast 120583g cmminus2 00123 0003Limit of detection (LOD) 120583gmLminus1 003 014Limit of quantification (LOQ) 120583gmLminus1 009 042Regression equation 119884lowastlowast

Intercept (119886) 00131 00198Slope (119887) 00905 03176Standard deviation of 119886 (119878

119886) 998 times 10minus2 998 times 10minus2

Standard deviation of 119887 (119878119887) 124 times 10minus2 398 times 10minus2

Regression coefficient (119903) 09971 09979lowastLimit of determination as the weight in 120583gmLminus1 of solution whichcorresponds to an absorbance of A = 0001 measured in a cuvette of cross-sectional area 1 cm2 and 119897 = 1 cmlowastlowast119884 = 119886 + 119887119883 where 119884 is the absorbance119883 is concentration in 120583gmLminus1 119886

is intercept and 119887 is slope

are the final chromogen and involve the following chemicalspecies

3H2O sdot P2O5sdot 13WO

3sdot 5MoO

3sdot 10H2O

3H2O sdot P2O5sdot 14WO

3sdot 4MoO

3sdot 10H2O

(2)

INH probably affects reduction of oxygen atoms fromtungstate andor molybdate in the F-C reagent by producingone or more possible reduced species which have character-istic intense blue color

The FFCmethod involves the redox reaction of INHwithferric chloride in the presence of potassium ferricyanideunder mild acidic conditions to produce a blue colour withmaximum absorption at 760 nm The first step in the colourdevelopment is the reduction of iron(III) of ferric chloride toiron(II) which subsequently reacts with ferricyanide to formPrussian blue

4 ISRN Analytical ChemistryAb

sorb

ance

1 1 FC-reagent (mL)

03

04

05

06

0 05 10 15 20 25 30

(a)

0

02

04

06

0 1 2 3 4 5

Abso

rban

ce

15 Na2CO3 (mL)

(b)

Abso

rban

ce

02

03

04

05

0 10 20 30 40 50 60

Time (min)

(c)

Figure 3 (a) Effect of volumes of F-C reagent (1 1) on the reactionproduct with INH (6 120583gmLminus1) in Na

2CO3solution for method A

(b) Effect of 20 Na2CO3on color formation (6120583gmLminus1 INH) for

method A (c) Reaction time and stability of colored species formethod A (6120583gmLminus1 INH)

7 Method Development

71 Spectral Characteristics The intensely blue colored prod-uct (molybdenum-tungsten mixed acid blue in FCR methodand Prussian blue in FFC method) formed in both methodsexhibited maximum absorption at 760 nm The absorptionspectra of the blue colored products and of the reagent blanksare shown in Figures 2(a) and 2(b)

Abso

rban

ce

035

040

045

050

FeCl3 (mL)05 10 15 20 25

(a)

Abso

rban

ce

030

035

040

045

050

Ferricyanide (mL)0 10 15 20 25

(b)

Abso

rban

ce

020

040

030

050

Reaction time (min)0 15 30 45 60 75 90 105 120

(c)

Figure 4 (a) Effect of volume of FeCl3for method B (15 120583gmLminus1

INH) (b) Effect of volume of ferricyanide formethod B (15120583gmLminus1INH) (c) Reaction time and stability of colored species for methodB (15 120583gmLminus1 INH)

72 Optimization of Experimental Variables A series ofpreliminary experiments necessary for rapid and quantitativeformation of colored products to achieve the maximumstability and sensitivity were performed Optimum conditionwas achieved by varying one parameter at a time whilekeeping other parameters constant and observing its effect onthe absorbance at 760 nm

ISRN Analytical Chemistry 5

Table 2 Evaluation of intraday and interday accuracy and precision

MethodINH Taken (120583gmLminus1) Intraday accuracy and precision (119899 = 7) Interday accuracy and precision (119899 = 5)

INH Founda (120583gmLminus1) RSDb

REc

INH found (120583gmLminus1) RSDb

REc

A30 296 124 089 307 234 21260 611 184 103 612 200 25390 914 156 152 915 167 200

B10 101 102 192 103 300 18515 147 201 115 154 267 12720 204 192 147 205 250 162

aMean value of 7 determinations brelative standard deviation () crelative error ()

Table 3 Method robustness and ruggedness expressed as intermediate precision

Method Nominal concentrationRobustness Ruggedness

Reaction timeslowast(119899 = 3)

Volumes of FCreagent

Volumesof ferricyanide

Interanalysts(119899 = 3)

Interinstruments(119899 = 3)

A30 145 169 mdash 220 23260 172 135 mdash 174 19290 210 171 mdash 244 257

B10 172 mdash 159 178 21215 184 mdash 161 211 16220 212 mdash 172 107 254

lowastReaction times were 10 plusmn 2min for both methods Volumes of FC regent added were 2 plusmn 02mL for method A volumes of ferricyanide added were 15 plusmn01mL for method B

73 Optimization for FCR Method

731 Effect of Concentration of F-C Reagent Several experi-ments were carried out to study the influence of F-C reagentconcentration on the color development and the obtainedresults are shown in Figure 3(a) It is apparent that 10 to50mL of reagent gave the maximum color intensity thus30mL of reagent was used throughout the investigation

732 Selection of Reaction Medium and Optimization of theBase To select a suitable medium for the reaction differentaqueous bases such as sodium hydroxide sodium carbon-ate or bicarbonate sodium acetate and sodium hydrogenphosphate were investigated Better results were obtainedwith sodium carbonate In order to determine the optimumconcentration of Na

2CO3 different volumes of 20 Na

2CO3

solution (0ndash5mL)were attempted at a constant concentrationof INH (6 120583gmLminus1) and the results of the observation areshown in Figure 3 It was found that different volumes rang-ing from 10 to 30mL of 20 Na

2CO3were optimum thus

20mL was used throughout the work (given in Figure 3(b))Maximum color development was obtained in 10min

after mixing the reactants and the color was stable for atleast 60min thereafter (Figure 3(c)) The sequence of orderof addition of the reactants had significant effect on theabsorbance value So the order used in the general procedureshould be followed for maximum absorbance

74 Optimization for FFC Method To optimize the concen-trations of ferricyanide and ferric chloride reagents differentvolumes of these reagents were used with a fixed concentra-tion of INH Constant absorbance was found with 10mL of05MFeCl

3and 15mLof 500120583gmLminus1 ferricyanide solutions

as shown in Figures 4(a) and 4(b)Colour development was complete in 10min and stable

for the next 90min (Figure 4(c)) Different results wereobtained when different order of addition of reactants wasfollowed The order of addition of reactants followed in therecommended procedure resulted in rapid color formationwith maximum sensitivity and stability

8 Method Validation

81 Linearity Sensitivity Limits of Detection and Quantifi-cation A linear correlation was found between absorbanceat 120582max and concentration of INH in the ranges given inTable 1 Regression analysis of the Beerrsquos law data using themethod of least squares was made to evaluate the slope (119887)intercept (119886) and correlation coefficient (119903) for each systemand the analytical results obtained from these investigationsare presented in Table 1 The optical characteristics such asBeerrsquos law limits molar absorptivity and Sandell sensitivityvalues are also given in Table 1 The high values of 120576 andlow values of Sandell sensitivity and LOD indicate the highsensitivity of the proposed methods

6 ISRN Analytical Chemistry

Table 4 Results of analysis of tablets by the proposed methods

Tablets analyzed Label claim (mgtablet)Foundlowast (percent of label claim plusmn SD)

Official method Proposed methodsMethod A Method B

Iskin-300 300 9986 plusmn 136

1015 plusmn 116 1012 plusmn 092

119905 = 206 119905 = 183

119865 = 137 119865 = 219

lowastMean value of five determinationsTabulated 119905-value at the 95 confidence level is 277Tabulated 119865-value at the 95 confidence level is 639

Table 5 Results of recovery study via standard addition method with tablet

Method Tablets studied INH in tablet 120583gmLminus1 Pure INH Added 120583gmLminus1 Total Found 120583gmLminus1 Pure INH recoveredlowastpercent plusmn SD

A Isokin-300296 15 441 9928 plusmn 203

296 30 601 1009 plusmn 312

296 45 756 1013 plusmn 299

B Isokin-300101 05 151 9985 plusmn 225

101 10 204 1013 plusmn 232

101 15 255 1017 plusmn 314

lowastMean value of three determinations

82 Intraday and Interday Precision and Accuracy The pre-cision and accuracy of the proposed method were studiedby repeating the experiment seven times within the dayto determine the repeatability (intraday precision) and fivetimes on different days to determine the intermediate preci-sion (interday precision) The assay was performed for threelevels of analyte in this method The results of this studyare summarized in Table 2 The percentage relative standarddeviation (RSD) values were le201 (intraday) and le300(interday) indicating good precision of themethod Accuracywas evaluated as percentage relative error (RE) between themeasured mean concentrations and taken concentrations ofINH and it was le253 demonstrating the high accuracy ofthe proposed method

83 Selectivity A selective study was performed to determinethe effect of matrix on the absorbance by analyzing theplacebo blank In the analysis of placebo blank solution theabsorbance in each case was equal to the absorbance ofblank which revealed no interference To assess the role ofthe inactive ingredients on the assay of INH the generalprocedure was applied on the synthetic mixture extract bytaking three different concentrations of INHwithin the rangeThe percentage recovery values were in the range 963ndash1023 and 954ndash1018 with RSD lt4 indicating clearly thenoninterference from the inactive ingredients in the assay ofINH

84 Robustness and Ruggedness The robustness of themethod was evaluated by making small incremental changesin the volume of reagent and reaction time and the effectsof the changes were studied by measuring the absorbance ofthe colored product The changes had negligible influence

on the results as revealed by small intermediate precisionvalues expressed as RSD (le254) Method ruggednesswas demonstrated having the analysis done by three analystsand also by a single analyst performing analysis on threedifferent instruments in the same laboratory Intermediateprecision values (RSD) in both instances indicated accept-able ruggedness These results are presented in Table 3

85 Application to Tablets The proposed methods wereapplied to the quantification of INH in commercial tabletsThe tablets were assayed by the official BP method [2] whichdescribes titration of the drug with potassium bromate inpresence of potassium bromide using methyl red indicatorThe results obtained by the proposed methods agree wellwith the claim and also are in agreement with those of theofficial method Statistical analysis of the results did notdetect any significant difference between the performance ofthe proposed method and reference method with respect toaccuracy and precision as revealed by the Studentrsquos 119905-valueand variance ratio 119865-value The results of assay are given inTable 4

86 Recovery Study To further assess the accuracy of themethod recovery experiment was performed by applying thestandard-addition technique The recovery was assessed bydetermining the agreement between the measured standardconcentration and added known concentration to the sampleThe test was done by spiking the preanalyzed tablet INHwithpure INH at three different levels (50 100 and 150) of thecontent present in the preparation and the total was found bythe proposedmethod Each test was repeated three timesTherecovery percentage values ranged between 9928 and 1017with standard deviation in the range 099ndash132 Closeness

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

4 ISRN Analytical ChemistryAb

sorb

ance

1 1 FC-reagent (mL)

03

04

05

06

0 05 10 15 20 25 30

(a)

0

02

04

06

0 1 2 3 4 5

Abso

rban

ce

15 Na2CO3 (mL)

(b)

Abso

rban

ce

02

03

04

05

0 10 20 30 40 50 60

Time (min)

(c)

Figure 3 (a) Effect of volumes of F-C reagent (1 1) on the reactionproduct with INH (6 120583gmLminus1) in Na

2CO3solution for method A

(b) Effect of 20 Na2CO3on color formation (6120583gmLminus1 INH) for

method A (c) Reaction time and stability of colored species formethod A (6120583gmLminus1 INH)

7 Method Development

71 Spectral Characteristics The intensely blue colored prod-uct (molybdenum-tungsten mixed acid blue in FCR methodand Prussian blue in FFC method) formed in both methodsexhibited maximum absorption at 760 nm The absorptionspectra of the blue colored products and of the reagent blanksare shown in Figures 2(a) and 2(b)

Abso

rban

ce

035

040

045

050

FeCl3 (mL)05 10 15 20 25

(a)

Abso

rban

ce

030

035

040

045

050

Ferricyanide (mL)0 10 15 20 25

(b)

Abso

rban

ce

020

040

030

050

Reaction time (min)0 15 30 45 60 75 90 105 120

(c)

Figure 4 (a) Effect of volume of FeCl3for method B (15 120583gmLminus1

INH) (b) Effect of volume of ferricyanide formethod B (15120583gmLminus1INH) (c) Reaction time and stability of colored species for methodB (15 120583gmLminus1 INH)

72 Optimization of Experimental Variables A series ofpreliminary experiments necessary for rapid and quantitativeformation of colored products to achieve the maximumstability and sensitivity were performed Optimum conditionwas achieved by varying one parameter at a time whilekeeping other parameters constant and observing its effect onthe absorbance at 760 nm

ISRN Analytical Chemistry 5

Table 2 Evaluation of intraday and interday accuracy and precision

MethodINH Taken (120583gmLminus1) Intraday accuracy and precision (119899 = 7) Interday accuracy and precision (119899 = 5)

INH Founda (120583gmLminus1) RSDb

REc

INH found (120583gmLminus1) RSDb

REc

A30 296 124 089 307 234 21260 611 184 103 612 200 25390 914 156 152 915 167 200

B10 101 102 192 103 300 18515 147 201 115 154 267 12720 204 192 147 205 250 162

aMean value of 7 determinations brelative standard deviation () crelative error ()

Table 3 Method robustness and ruggedness expressed as intermediate precision

Method Nominal concentrationRobustness Ruggedness

Reaction timeslowast(119899 = 3)

Volumes of FCreagent

Volumesof ferricyanide

Interanalysts(119899 = 3)

Interinstruments(119899 = 3)

A30 145 169 mdash 220 23260 172 135 mdash 174 19290 210 171 mdash 244 257

B10 172 mdash 159 178 21215 184 mdash 161 211 16220 212 mdash 172 107 254

lowastReaction times were 10 plusmn 2min for both methods Volumes of FC regent added were 2 plusmn 02mL for method A volumes of ferricyanide added were 15 plusmn01mL for method B

73 Optimization for FCR Method

731 Effect of Concentration of F-C Reagent Several experi-ments were carried out to study the influence of F-C reagentconcentration on the color development and the obtainedresults are shown in Figure 3(a) It is apparent that 10 to50mL of reagent gave the maximum color intensity thus30mL of reagent was used throughout the investigation

732 Selection of Reaction Medium and Optimization of theBase To select a suitable medium for the reaction differentaqueous bases such as sodium hydroxide sodium carbon-ate or bicarbonate sodium acetate and sodium hydrogenphosphate were investigated Better results were obtainedwith sodium carbonate In order to determine the optimumconcentration of Na

2CO3 different volumes of 20 Na

2CO3

solution (0ndash5mL)were attempted at a constant concentrationof INH (6 120583gmLminus1) and the results of the observation areshown in Figure 3 It was found that different volumes rang-ing from 10 to 30mL of 20 Na

2CO3were optimum thus

20mL was used throughout the work (given in Figure 3(b))Maximum color development was obtained in 10min

after mixing the reactants and the color was stable for atleast 60min thereafter (Figure 3(c)) The sequence of orderof addition of the reactants had significant effect on theabsorbance value So the order used in the general procedureshould be followed for maximum absorbance

74 Optimization for FFC Method To optimize the concen-trations of ferricyanide and ferric chloride reagents differentvolumes of these reagents were used with a fixed concentra-tion of INH Constant absorbance was found with 10mL of05MFeCl

3and 15mLof 500120583gmLminus1 ferricyanide solutions

as shown in Figures 4(a) and 4(b)Colour development was complete in 10min and stable

for the next 90min (Figure 4(c)) Different results wereobtained when different order of addition of reactants wasfollowed The order of addition of reactants followed in therecommended procedure resulted in rapid color formationwith maximum sensitivity and stability

8 Method Validation

81 Linearity Sensitivity Limits of Detection and Quantifi-cation A linear correlation was found between absorbanceat 120582max and concentration of INH in the ranges given inTable 1 Regression analysis of the Beerrsquos law data using themethod of least squares was made to evaluate the slope (119887)intercept (119886) and correlation coefficient (119903) for each systemand the analytical results obtained from these investigationsare presented in Table 1 The optical characteristics such asBeerrsquos law limits molar absorptivity and Sandell sensitivityvalues are also given in Table 1 The high values of 120576 andlow values of Sandell sensitivity and LOD indicate the highsensitivity of the proposed methods

6 ISRN Analytical Chemistry

Table 4 Results of analysis of tablets by the proposed methods

Tablets analyzed Label claim (mgtablet)Foundlowast (percent of label claim plusmn SD)

Official method Proposed methodsMethod A Method B

Iskin-300 300 9986 plusmn 136

1015 plusmn 116 1012 plusmn 092

119905 = 206 119905 = 183

119865 = 137 119865 = 219

lowastMean value of five determinationsTabulated 119905-value at the 95 confidence level is 277Tabulated 119865-value at the 95 confidence level is 639

Table 5 Results of recovery study via standard addition method with tablet

Method Tablets studied INH in tablet 120583gmLminus1 Pure INH Added 120583gmLminus1 Total Found 120583gmLminus1 Pure INH recoveredlowastpercent plusmn SD

A Isokin-300296 15 441 9928 plusmn 203

296 30 601 1009 plusmn 312

296 45 756 1013 plusmn 299

B Isokin-300101 05 151 9985 plusmn 225

101 10 204 1013 plusmn 232

101 15 255 1017 plusmn 314

lowastMean value of three determinations

82 Intraday and Interday Precision and Accuracy The pre-cision and accuracy of the proposed method were studiedby repeating the experiment seven times within the dayto determine the repeatability (intraday precision) and fivetimes on different days to determine the intermediate preci-sion (interday precision) The assay was performed for threelevels of analyte in this method The results of this studyare summarized in Table 2 The percentage relative standarddeviation (RSD) values were le201 (intraday) and le300(interday) indicating good precision of themethod Accuracywas evaluated as percentage relative error (RE) between themeasured mean concentrations and taken concentrations ofINH and it was le253 demonstrating the high accuracy ofthe proposed method

83 Selectivity A selective study was performed to determinethe effect of matrix on the absorbance by analyzing theplacebo blank In the analysis of placebo blank solution theabsorbance in each case was equal to the absorbance ofblank which revealed no interference To assess the role ofthe inactive ingredients on the assay of INH the generalprocedure was applied on the synthetic mixture extract bytaking three different concentrations of INHwithin the rangeThe percentage recovery values were in the range 963ndash1023 and 954ndash1018 with RSD lt4 indicating clearly thenoninterference from the inactive ingredients in the assay ofINH

84 Robustness and Ruggedness The robustness of themethod was evaluated by making small incremental changesin the volume of reagent and reaction time and the effectsof the changes were studied by measuring the absorbance ofthe colored product The changes had negligible influence

on the results as revealed by small intermediate precisionvalues expressed as RSD (le254) Method ruggednesswas demonstrated having the analysis done by three analystsand also by a single analyst performing analysis on threedifferent instruments in the same laboratory Intermediateprecision values (RSD) in both instances indicated accept-able ruggedness These results are presented in Table 3

85 Application to Tablets The proposed methods wereapplied to the quantification of INH in commercial tabletsThe tablets were assayed by the official BP method [2] whichdescribes titration of the drug with potassium bromate inpresence of potassium bromide using methyl red indicatorThe results obtained by the proposed methods agree wellwith the claim and also are in agreement with those of theofficial method Statistical analysis of the results did notdetect any significant difference between the performance ofthe proposed method and reference method with respect toaccuracy and precision as revealed by the Studentrsquos 119905-valueand variance ratio 119865-value The results of assay are given inTable 4

86 Recovery Study To further assess the accuracy of themethod recovery experiment was performed by applying thestandard-addition technique The recovery was assessed bydetermining the agreement between the measured standardconcentration and added known concentration to the sampleThe test was done by spiking the preanalyzed tablet INHwithpure INH at three different levels (50 100 and 150) of thecontent present in the preparation and the total was found bythe proposedmethod Each test was repeated three timesTherecovery percentage values ranged between 9928 and 1017with standard deviation in the range 099ndash132 Closeness

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

ISRN Analytical Chemistry 5

Table 2 Evaluation of intraday and interday accuracy and precision

MethodINH Taken (120583gmLminus1) Intraday accuracy and precision (119899 = 7) Interday accuracy and precision (119899 = 5)

INH Founda (120583gmLminus1) RSDb

REc

INH found (120583gmLminus1) RSDb

REc

A30 296 124 089 307 234 21260 611 184 103 612 200 25390 914 156 152 915 167 200

B10 101 102 192 103 300 18515 147 201 115 154 267 12720 204 192 147 205 250 162

aMean value of 7 determinations brelative standard deviation () crelative error ()

Table 3 Method robustness and ruggedness expressed as intermediate precision

Method Nominal concentrationRobustness Ruggedness

Reaction timeslowast(119899 = 3)

Volumes of FCreagent

Volumesof ferricyanide

Interanalysts(119899 = 3)

Interinstruments(119899 = 3)

A30 145 169 mdash 220 23260 172 135 mdash 174 19290 210 171 mdash 244 257

B10 172 mdash 159 178 21215 184 mdash 161 211 16220 212 mdash 172 107 254

lowastReaction times were 10 plusmn 2min for both methods Volumes of FC regent added were 2 plusmn 02mL for method A volumes of ferricyanide added were 15 plusmn01mL for method B

73 Optimization for FCR Method

731 Effect of Concentration of F-C Reagent Several experi-ments were carried out to study the influence of F-C reagentconcentration on the color development and the obtainedresults are shown in Figure 3(a) It is apparent that 10 to50mL of reagent gave the maximum color intensity thus30mL of reagent was used throughout the investigation

732 Selection of Reaction Medium and Optimization of theBase To select a suitable medium for the reaction differentaqueous bases such as sodium hydroxide sodium carbon-ate or bicarbonate sodium acetate and sodium hydrogenphosphate were investigated Better results were obtainedwith sodium carbonate In order to determine the optimumconcentration of Na

2CO3 different volumes of 20 Na

2CO3

solution (0ndash5mL)were attempted at a constant concentrationof INH (6 120583gmLminus1) and the results of the observation areshown in Figure 3 It was found that different volumes rang-ing from 10 to 30mL of 20 Na

2CO3were optimum thus

20mL was used throughout the work (given in Figure 3(b))Maximum color development was obtained in 10min

after mixing the reactants and the color was stable for atleast 60min thereafter (Figure 3(c)) The sequence of orderof addition of the reactants had significant effect on theabsorbance value So the order used in the general procedureshould be followed for maximum absorbance

74 Optimization for FFC Method To optimize the concen-trations of ferricyanide and ferric chloride reagents differentvolumes of these reagents were used with a fixed concentra-tion of INH Constant absorbance was found with 10mL of05MFeCl

3and 15mLof 500120583gmLminus1 ferricyanide solutions

as shown in Figures 4(a) and 4(b)Colour development was complete in 10min and stable

for the next 90min (Figure 4(c)) Different results wereobtained when different order of addition of reactants wasfollowed The order of addition of reactants followed in therecommended procedure resulted in rapid color formationwith maximum sensitivity and stability

8 Method Validation

81 Linearity Sensitivity Limits of Detection and Quantifi-cation A linear correlation was found between absorbanceat 120582max and concentration of INH in the ranges given inTable 1 Regression analysis of the Beerrsquos law data using themethod of least squares was made to evaluate the slope (119887)intercept (119886) and correlation coefficient (119903) for each systemand the analytical results obtained from these investigationsare presented in Table 1 The optical characteristics such asBeerrsquos law limits molar absorptivity and Sandell sensitivityvalues are also given in Table 1 The high values of 120576 andlow values of Sandell sensitivity and LOD indicate the highsensitivity of the proposed methods

6 ISRN Analytical Chemistry

Table 4 Results of analysis of tablets by the proposed methods

Tablets analyzed Label claim (mgtablet)Foundlowast (percent of label claim plusmn SD)

Official method Proposed methodsMethod A Method B

Iskin-300 300 9986 plusmn 136

1015 plusmn 116 1012 plusmn 092

119905 = 206 119905 = 183

119865 = 137 119865 = 219

lowastMean value of five determinationsTabulated 119905-value at the 95 confidence level is 277Tabulated 119865-value at the 95 confidence level is 639

Table 5 Results of recovery study via standard addition method with tablet

Method Tablets studied INH in tablet 120583gmLminus1 Pure INH Added 120583gmLminus1 Total Found 120583gmLminus1 Pure INH recoveredlowastpercent plusmn SD

A Isokin-300296 15 441 9928 plusmn 203

296 30 601 1009 plusmn 312

296 45 756 1013 plusmn 299

B Isokin-300101 05 151 9985 plusmn 225

101 10 204 1013 plusmn 232

101 15 255 1017 plusmn 314

lowastMean value of three determinations

82 Intraday and Interday Precision and Accuracy The pre-cision and accuracy of the proposed method were studiedby repeating the experiment seven times within the dayto determine the repeatability (intraday precision) and fivetimes on different days to determine the intermediate preci-sion (interday precision) The assay was performed for threelevels of analyte in this method The results of this studyare summarized in Table 2 The percentage relative standarddeviation (RSD) values were le201 (intraday) and le300(interday) indicating good precision of themethod Accuracywas evaluated as percentage relative error (RE) between themeasured mean concentrations and taken concentrations ofINH and it was le253 demonstrating the high accuracy ofthe proposed method

83 Selectivity A selective study was performed to determinethe effect of matrix on the absorbance by analyzing theplacebo blank In the analysis of placebo blank solution theabsorbance in each case was equal to the absorbance ofblank which revealed no interference To assess the role ofthe inactive ingredients on the assay of INH the generalprocedure was applied on the synthetic mixture extract bytaking three different concentrations of INHwithin the rangeThe percentage recovery values were in the range 963ndash1023 and 954ndash1018 with RSD lt4 indicating clearly thenoninterference from the inactive ingredients in the assay ofINH

84 Robustness and Ruggedness The robustness of themethod was evaluated by making small incremental changesin the volume of reagent and reaction time and the effectsof the changes were studied by measuring the absorbance ofthe colored product The changes had negligible influence

on the results as revealed by small intermediate precisionvalues expressed as RSD (le254) Method ruggednesswas demonstrated having the analysis done by three analystsand also by a single analyst performing analysis on threedifferent instruments in the same laboratory Intermediateprecision values (RSD) in both instances indicated accept-able ruggedness These results are presented in Table 3

85 Application to Tablets The proposed methods wereapplied to the quantification of INH in commercial tabletsThe tablets were assayed by the official BP method [2] whichdescribes titration of the drug with potassium bromate inpresence of potassium bromide using methyl red indicatorThe results obtained by the proposed methods agree wellwith the claim and also are in agreement with those of theofficial method Statistical analysis of the results did notdetect any significant difference between the performance ofthe proposed method and reference method with respect toaccuracy and precision as revealed by the Studentrsquos 119905-valueand variance ratio 119865-value The results of assay are given inTable 4

86 Recovery Study To further assess the accuracy of themethod recovery experiment was performed by applying thestandard-addition technique The recovery was assessed bydetermining the agreement between the measured standardconcentration and added known concentration to the sampleThe test was done by spiking the preanalyzed tablet INHwithpure INH at three different levels (50 100 and 150) of thecontent present in the preparation and the total was found bythe proposedmethod Each test was repeated three timesTherecovery percentage values ranged between 9928 and 1017with standard deviation in the range 099ndash132 Closeness

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

6 ISRN Analytical Chemistry

Table 4 Results of analysis of tablets by the proposed methods

Tablets analyzed Label claim (mgtablet)Foundlowast (percent of label claim plusmn SD)

Official method Proposed methodsMethod A Method B

Iskin-300 300 9986 plusmn 136

1015 plusmn 116 1012 plusmn 092

119905 = 206 119905 = 183

119865 = 137 119865 = 219

lowastMean value of five determinationsTabulated 119905-value at the 95 confidence level is 277Tabulated 119865-value at the 95 confidence level is 639

Table 5 Results of recovery study via standard addition method with tablet

Method Tablets studied INH in tablet 120583gmLminus1 Pure INH Added 120583gmLminus1 Total Found 120583gmLminus1 Pure INH recoveredlowastpercent plusmn SD

A Isokin-300296 15 441 9928 plusmn 203

296 30 601 1009 plusmn 312

296 45 756 1013 plusmn 299

B Isokin-300101 05 151 9985 plusmn 225

101 10 204 1013 plusmn 232

101 15 255 1017 plusmn 314

lowastMean value of three determinations

82 Intraday and Interday Precision and Accuracy The pre-cision and accuracy of the proposed method were studiedby repeating the experiment seven times within the dayto determine the repeatability (intraday precision) and fivetimes on different days to determine the intermediate preci-sion (interday precision) The assay was performed for threelevels of analyte in this method The results of this studyare summarized in Table 2 The percentage relative standarddeviation (RSD) values were le201 (intraday) and le300(interday) indicating good precision of themethod Accuracywas evaluated as percentage relative error (RE) between themeasured mean concentrations and taken concentrations ofINH and it was le253 demonstrating the high accuracy ofthe proposed method

83 Selectivity A selective study was performed to determinethe effect of matrix on the absorbance by analyzing theplacebo blank In the analysis of placebo blank solution theabsorbance in each case was equal to the absorbance ofblank which revealed no interference To assess the role ofthe inactive ingredients on the assay of INH the generalprocedure was applied on the synthetic mixture extract bytaking three different concentrations of INHwithin the rangeThe percentage recovery values were in the range 963ndash1023 and 954ndash1018 with RSD lt4 indicating clearly thenoninterference from the inactive ingredients in the assay ofINH

84 Robustness and Ruggedness The robustness of themethod was evaluated by making small incremental changesin the volume of reagent and reaction time and the effectsof the changes were studied by measuring the absorbance ofthe colored product The changes had negligible influence

on the results as revealed by small intermediate precisionvalues expressed as RSD (le254) Method ruggednesswas demonstrated having the analysis done by three analystsand also by a single analyst performing analysis on threedifferent instruments in the same laboratory Intermediateprecision values (RSD) in both instances indicated accept-able ruggedness These results are presented in Table 3

85 Application to Tablets The proposed methods wereapplied to the quantification of INH in commercial tabletsThe tablets were assayed by the official BP method [2] whichdescribes titration of the drug with potassium bromate inpresence of potassium bromide using methyl red indicatorThe results obtained by the proposed methods agree wellwith the claim and also are in agreement with those of theofficial method Statistical analysis of the results did notdetect any significant difference between the performance ofthe proposed method and reference method with respect toaccuracy and precision as revealed by the Studentrsquos 119905-valueand variance ratio 119865-value The results of assay are given inTable 4

86 Recovery Study To further assess the accuracy of themethod recovery experiment was performed by applying thestandard-addition technique The recovery was assessed bydetermining the agreement between the measured standardconcentration and added known concentration to the sampleThe test was done by spiking the preanalyzed tablet INHwithpure INH at three different levels (50 100 and 150) of thecontent present in the preparation and the total was found bythe proposedmethod Each test was repeated three timesTherecovery percentage values ranged between 9928 and 1017with standard deviation in the range 099ndash132 Closeness

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

ISRN Analytical Chemistry 7Ta

ble6Com

paris

onof

perfo

rmance

characteris

ticso

fthe

prop

osed

metho

dswith

thee

xistingmetho

ds

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

1Ep

ichloroh

ydrin

Measuremento

fpurplec

olored

coup

led

complex

405

2ndash22

(051times

104 )

Longer

reactio

ntim

eand

heating

[45]

hydroxyphenacylch

lorid

e402

20ndash120

(010times10

4 )

2Ethylvanillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

410

2ndash16

(71times

103 )

mdash[46]

3Va

nillin

Measuremento

fyellowcolored

hydrazon

ecom

plex

405

1ndash12

mdash[47]

4Isatin

Measuremento

fyellowcolored

hydrazon

ecom

plex

340

0ndash32

(12times10

4 )

Requ

iresc

lose

pHcontroland

40min

standing

timemeasurementata

shorter

wavele

ngth

[48]

5

Sodium

12-naphtho

quinon

e-4-

sulfo

nateand

cetyltrim

ethylammon

ium

brom

ide

Absorbance

ofcond

ensatio

nprod

uct

measured

500

20ndash

56

Employsa

costlyreagent

[49]

6

Chloranilic

acid

Measuremento

fcharge-transfe

rcom

plex

500

137ndash82

Use

oforganics

olventrequiresh

eatin

gste

p

[50]

Tetracyano

ethylene

480

685ndash3427

23-dichloro-56-dicyano

-14

-benzoqu

inon

e580

1096ndash

2193

Thym

olblue

Measuremento

fion

-associatecomplex

390

685ndash4

113

Requ

ireclo

sepH

controlliq

uid-liq

uid

extractio

nste

pBrom

opheno

lblue

410

137ndash685

Brom

ocresolgreen

320

137ndash822

72-Hydroxy-14-

naptho

quinon

eMeasuremento

fderivatized

prod

uct

365

5ndash25

Requ

iresc

lose

pHcontroluseo

fno

naqu

eous

mediumm

easurementata

shorterw

avele

ngth

[51]

812-Naphtho

quinon

e-4-

sulfo

nate

Measuremento

fpinkcoloredcond

ensed

prod

uct

495

05ndash30

(118times10

4 )Re

quire

sclose

pHcontrol

[52]

96-Methyl-2

-pyrid

inecarbo

xaldehyde

Measuremento

fhydrazone

deriv

ative

328

2ndash16

Measurementatlow

eranalytical

wavele

ngth

[53]

109-Ch

loroacrid

ine

Measuremento

fabsorbanceo

fderiv

atized

complex

500

mdashRe

quire

sheatin

gste

ptim

econ

suming

[54]

11441015840-M

ethylene-bis-m-

nitro

aniline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

495

01ndash15

(563times10

4 )Re

quire

slow

temperature

[55]

12441015840-Sulph

onyldianiline

Measuremento

fpurplec

olored

diazo-coup

ledcomplex

440

335

05ndash20

(572times10

4 )Re

quire

slow

temperature

[56]

13Tiron-NaIO

4Measuremento

fred-colored

oxidative-coup

ledprod

uct

507

10ndash15

(184times10

4 )Use

ofmultireactionsyste

mste

p[57]

14Tiron-KI

O4

Measuremento

fred-colored

oxidative-coup

ledprod

uct

505

15ndash18

(177times10

4 )Use

ofmultistepreactio

nsyste

m[58]

15Uranylacetate

Measuremento

fyellow-colored

uranyl

isonicotin

oyld

ithiocarbazatec

omplex

410

mdashMultistepreactio

ntim

econ

suming

[59]

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

8 ISRN Analytical Chemistry

Table6Con

tinued

SlN

oRe

agents

used

Metho

dology

120582max

(nm)

Linear

range

(120583gm

Lminus1)

(120576=Lmolcm)

Remarks

References

16Neocuproine

Measuremento

fredox

complex

454

03ndash35

Requ

iresc

lose

pHcontrolam

ultistep

reactio

n[60]

17Ro

sebeng

alMeasuremento

facetone-chloroform

extractableion

-pairc

omplex

555

28ndash56

Requ

irestim

econ

sumingandtediou

sextractio

nste

puseo

forganicsolvent

[61]

18lowastNBS

Measuremento

fstarch-iodine

Com

plex

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[62]

19lowastNBS

Absorbance

ofsta

rch-iodine

complex

measured

572

01ndash34

Multistepreactio

nem

ploysa

nun

stable

oxidant

[63]

20Po

tassium

ferricyanide

Measuremento

fPrussianblue

735

004ndash8

(392times10

4 )mdash

[64]

213-(2-Pyridyl)-56-diph

enyl-

124-tr

iazine

Measuremento

fredox

complex

558

062ndash6

15Multistepreactio

nrequ

iresa

nexpensive

reagent

[65]

22(a)lowastFC

reagent

Measuremento

fmolyb

denu

m-tu

ngsten

mixed

acid

blue

absorban

cein

sodium

carbon

atem

edium

760

05ndash

10(112times10

4 )Simplerapid

sensitiveu

sesa

queous

medium

andextractio

nfreen

ouseo

forganics

olventb

ased

onas

ingles

tep

reactio

n

Presentw

ork

(b)F

erricc

hloridea

ndpo

tassium

ferricyanide

Measuremento

fabsorba

nceo

fPrussian

blue

inacid

medium

760

02ndash

30

(455times10

4 )lowastNBS

-N-bromosuccinimideKM

nO4-po

tassium

perm

anganateFCreagent-F

olin-C

iocalteureagent

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

ISRN Analytical Chemistry 9

of the results to 100 showed the fairly good accuracy of themethod The results are shown in Table 5

9 Conclusion

Two simple rapid selective and sensitive methods havebeen proposed for the assay of INH in bulk drugs and intablets The methods are based on the well-characterized andestablished redox and complexation reactions and use verycommon and inexpensive chemicals and easily accessibleinstrument The procedures described here are easily carriedout and much simpler than the reported methods for INH(Table 6) but have been demonstrated to be more sensitivein terms of linear dynamic range and sensitivity parametersThe methods were applied successfully to the assay of INHin tablets without interferences from the common excipientsThe proposed methods are suitable for isoniazid determina-tion in bulk drug and pharmaceuticals hence these methodscan be used in quality control laboratories

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors wish to thank the quality control managerCipla Ltd Bangalore India for gifting pure isoniazid andthe authorities of the University of Mysore Mysore forpermission and facilities

References

[1] Indian Pharmacopoea Vol I The controller of PublicationsGovt of India New Delhi India 4th edition 1996

[2] British Pharmacopoeia Volume I and II Her Majestyrsquos Sta-tionery Office London UK 2009

[3] The United States Pharmacopoeia XXIV Revision The NationalFormulary XIX Rockville USP Convention 2000

[4] P Nagendra H S Yathirajan K N Mohana and K SRangappa ldquoOxidation of isoniazid and glutathione withbromamine-Trdquo Journal of the Indian Chemical Society vol 79no 1 pp 75ndash78 2002

[5] C R Raju H S Yathirajan K S Rangappa K N Mohanaand K M L Rai ldquoOxidimetric determination of isoniazid andamino acids with bromamine-B in buffer mediumrdquo OxidationCommunications vol 24 no 3 pp 393ndash399 2001

[6] B M Mohan H S Yathirajan R Rangaswamy and JJayarama ldquoDetermination of ascorbic acid and isoniazidwith n-bromosuccinimiderdquo Indian Journal of Pharmaceutical Sciencesvol 46 no 4 pp 156ndash158 1984

[7] A Das and K S Boparai ldquoTitration of thiacetazone andisoniazid with sodium methoxide in non-aqueous mediumrdquoTalanta vol 29 no 1 pp 57ndash60 1982

[8] U P Azad and V Ganesan ldquoEfficient electrocatalytic oxidationand selective determination of isoniazid by Fe(tmphen)2+

3-

exchanged Nafion-modified electroderdquo Journal of Solid StateElectrochemistry vol 16 no 9 pp 2907ndash2911 2012

[9] S Shahrokhian and E Asadian ldquoSimultaneous voltammetricdetermination of ascorbic acid acetaminophen and isoniazidusing thionine immobilized multi-walled carbon nanotubemodified carbon paste electroderdquo Electrochimica Acta vol 55no 3 pp 666ndash672 2010

[10] G Yang C Wang R Zhang C Wang Q Qu and X HuldquoPoly(amidosulfonic acid) modified glassy carbon electrode fordetermination of isoniazid in pharmaceuticalsrdquoBioelectrochem-istry vol 73 no 1 pp 37ndash42 2008

[11] Z-N Gao X-X Han H-Q Yao B Liang andW-Y Liu ldquoElec-trochemical oxidation of isoniazid catalyzed by (FcM)TMA atthe platinum electrode and its practical analytical applicationrdquoAnalytical and Bioanalytical Chemistry vol 385 no 7 pp 1324ndash1329 2006

[12] M R Majidi A Jouyban and K Asadpour-Zeynali ldquoVoltam-metric behavior and determination of isoniazid in pharmaceu-ticals by using overoxidized polypyrrole glassy carbonmodifiedelectroderdquo Journal of Electroanalytical Chemistry vol 589 no 1pp 32ndash37 2006

[13] T Wahdan ldquoVoltammetric method for the simultaneous deter-mination of rifampicin and isoniazid in pharmaceutical formu-lationsrdquo Chemia Analityczna vol 50 no 2 pp 457ndash464 2005

[14] I Pasakova M Gladziszova J Charvatova J Stariat J Klimesand P Kovarıkova ldquoUse of different stationary phases forseparation of isoniazid its metabolites and vitamin B6 formsrdquoJournal of Separation Science vol 34 no 12 pp 1357ndash1365 2011

[15] M Gajendiran andM M Abdul Kamal Nazer ldquoPotentiometricback titration of isoniazid in pharmaceutical dosage formsusing copper based mercury film electroderdquo Journal of theKorean Chemical Society vol 55 no 4 pp 620ndash625 2011

[16] L Lakshmi ldquoPotentiotitrimetric determination of isoniazidusing twin copper based mercury film electroderdquo Asian Journalof Chemistry vol 22 no 8 pp 6067ndash6076 2010

[17] P Riyazuddin and M M Abdul Kamal Nazer ldquoIndirect poten-tiometric titration of isoniazid in pharmaceutical dosage formsusing a copper based mercury film electroderdquo Indian Journal ofPharmaceutical Sciences vol 60 no 3 pp 158ndash161 1998

[18] MA Koupparis andT PHadjiioannou ldquoIndirect potentiomet-ric determination of hydrazine isoniazid sulphide and thiosul-phate with a chloramine-T ion-selective electroderdquoTalanta vol25 no 8 pp 477ndash480 1978

[19] P V Krishna Rao and G Bala Bhaskara Rao ldquoA potentiometricprocedure for the assay of isonicotinic acid hydrazide (isoni-azid)rdquoThe Analyst vol 96 no 147 pp 712ndash715 1971

[20] WC Chen BUnnikrishnan and SMChen ldquoElectrochemicaloxidation and amperometric determination of isoniazid atfunctionalized multiwalled carbon nanotube modified elec-troderdquo International Journal of Electrochemical Science vol 7no 10 pp 9138ndash9149 2012

[21] M S M Quintino and L Angnes ldquoFast BIA-amperometricdetermination of isoniazid in tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 42 no 3 pp 400ndash404 2006

[22] T Hossein H Yahya and T Afsaneh ldquoA selective and simplemethod for isoniazid spectrofluorimetric determination basedon the oxidation by cerium(IV)rdquo Asian Journal of Biochemicaland Pharmaceutical Research vol 1 no 2 pp 712ndash718 2011

[23] J A Garcıa Bautista J V Garcıa Mateo and J MartınezCalatayud ldquoSpectrofluorimetric determination of iproniazidand isoniazid in a FIA system provided with a solid-phasereactorrdquo Analytical Letters vol 31 no 7 pp 1209ndash1218 1998

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

10 ISRN Analytical Chemistry

[24] W Bowan W Zhihua X Zhonghua et al ldquoA novel molecu-larty imprinted electrochemiluminescence sensor for isoniaziddetectionrdquoThe Analyst vol 137 no 16 pp 3644ndash3652 2012

[25] J Xi B Shi X Ai and Z He ldquoChemiluminescence detection ofisoniazid using Ru(phen)2+

3-isoniazid-Ce(IV) systemrdquo Journal

of Pharmaceutical and Biomedical Analysis vol 36 no 1 pp237ndash241 2004

[26] A Safavi M A Karimi and M R H Nezhad ldquoFlow injectiondetermination of isoniazid using N-bromosuccinimide- andN-chlorosuccinimide-luminol chemiluminescence systemsrdquo Jour-nal of Pharmaceutical and Biomedical Analysis vol 30 no 5 pp1499ndash1506 2003

[27] S Zhang and H Li ldquoFlow-injection chemiluminescence sensorfor the determination of isoniazidrdquoAnalytica Chimica Acta vol444 no 2 pp 287ndash294 2001

[28] X Zheng Z Guo and Z Zhang ldquoFlow-injection electrogen-erated chemiluminescence determination of isoniazid usingluminolrdquo Analytical Sciences vol 17 no 9 pp 1095ndash1099 2001

[29] Y Huang and Z Zhang ldquoFlow injection chemiluminescentanalysis of isoniazid by direct hexacyanoferrate(III) oxidationrdquoAnalytical Letters vol 34 no 10 pp 1703ndash1710 2001

[30] Y Huang Z Zhang D Zhang and J Lv ldquoA flow injectionchemiluminescence system for the determination of isoniazidrdquoFreseniusrsquo Journal of Analytical Chemistry vol 368 no 4 pp429ndash431 2000

[31] B Li Z Zhang X Zheng and C Xu ldquoFlow injection chemi-luminescence determination of isoniazid using on-line electro-generated manganese(III) as oxidantrdquo Microchemical Journalvol 63 no 3 pp 374ndash380 1999

[32] X Zheng and Z Zhang ldquoFlow-injection chemiluminescencedetermination of isoniazid using on-line electrogenerated BrO-as an oxidantrdquoThe Analyst vol 124 no 5 pp 763ndash766 1999

[33] J Huang C Zhang and Z Zhang ldquoFlow injection chemilu-minescence determination of isoniazid with electrogeneratedhypochloriterdquo Freseniusrsquo Journal of Analytical Chemistry vol363 no 1 pp 126ndash128 1999

[34] J Ayyappan P Umapathi and S Darlin Quine ldquoDevelopmentand validation of a stability indicating high-performance liquidchromatography (HPLC) method for the estimation of iso-niazid and its related substances in fixed dose combinationof isoniazid and ethambutol hydrochloride tabletsrdquo AfricanJournal of Pharmacy and Pharmacology vol 5 no 12 pp 1513ndash1521 2011

[35] S Gunasekaran and E Sailatha ldquoEstimation of pyrazinamideisoniazid and rifampicin in pharmaceutical formulations byhigh performance liquid chromatography methodrdquo Asian Jour-nal of Chemistry vol 21 no 5 pp 3561ndash3566 2009

[36] B D Glass S Agatonovic-Kustrin Y-J Chen and M HWisch ldquoOptimization of a stability-indicating HPLC methodfor the simultaneous determination of rifampicin isoniazidand pyrazinamide in a fixed-dose combination using artificialneural networksrdquo Journal of Chromatographic Science vol 45no 1 pp 38ndash44 2007

[37] V D Gupta and A Sood ldquoChemical stability of isoniazid in anoral liquid dosage formrdquo International Journal of Pharmacy andCompounding vol 9 no 2 pp 165ndash166 2005

[38] M Y Khuhawar and L A Zardari ldquoEthyl chloroformate as aderivatizing reagent for the gas chromatographic determinationof isoniazid and hydrazine in pharmaceutical preparationsrdquoAnalytical Sciences vol 24 no 11 pp 1493ndash1496 2008

[39] M Y Khuhawar L A Zardari and A J Laghari ldquoCapillary gaschromatographic determination of isoniazid in pharmaceutical

preparation by pre-column derivatization with acetylacetonerdquoAsian Journal of Chemistry vol 20 no 8 pp 5997ndash6006 2008

[40] M Y Khuhawar and L A Zardari ldquoCapillary gas chromato-graphic determination of isoniazid in pharmaceutical prepa-rations and blood precolumn derivazation with trifluoroacety-lacetonerdquo Journal of Food and Drug Analysis vol 14 no 4 pp323ndash328 2006

[41] H Bhutani S Singh S Vir et al ldquoLC and LC-MS study ofstress decomposition behaviour of isoniazid and establishmentof validated stability-indicating assay methodrdquo Journal of Phar-maceutical and Biomedical Analysis vol 43 no 4 pp 1213ndash12202007

[42] Y Liu Z Fu and L Wang ldquoCapillary electrophoresis analysisof isoniazid using luminol-periodate potassium chemilumines-cence systemrdquo Luminescence vol 26 no 6 pp 397ndash402 2011

[43] X Zhang Y Xuan A Sun Y Lv and X Hou ldquoSimultane-ous determination of isoniazid and p-aminosalicylic acid bycapillary electrophoresis using chemiluminescence detectionrdquoLuminescence vol 24 no 4 pp 243ndash249 2009

[44] R Driouich T Takayanagi M Mitsuko O Oshima andS Motomizu ldquoInvestigation of salicylaldehyde-5-sulfonate asa precolumn derivatizing agent for the determination of n-alkane diamines lysine diaminopimelic acid and isoniazid bycapillary zone electrophoresisrdquo Journal of Pharmaceutical andBiomedical Analysis vol 30 no 5 pp 1523ndash1530 2003

[45] D N Shetty N Badiyadka and S Samshuddin ldquoNovel reagentsfor the spectrophotometric determination of isoniazidrdquo ISRNSpectroscopy vol 2012 Article ID 869493 5 pages 2012

[46] R Kashyap E V S Subrahmanyam and A R SharbarayamldquoDevelopment and validation of new colorimetric methodfor the estimation of isoniazid in bulk and dosage formrdquoInternational Journal of Pharmacy and Pharmaceutical Sciencesvol 4 no 3 pp 688ndash695 2012

[47] E F Oga ldquoSpectrophotometric determination of isoniazid inpure and pharmaceutical formulations using vanillinrdquo Interna-tional Journal of Pharmacy and Pharmaceutical Sciences vol 2no 1 pp 55ndash58 2010

[48] M N Abbas and A M A Homoda ldquoSpectrophotomet-ric determination of isoniazid in presence of rifampicin insome pharmaceutical preparations and urine using isatin as areagentrdquo Egyptian Journal of Chemistry vol 46 no 1 pp 57ndash692003

[49] P Nagaraja K C SrinivasaMurthy andH S Yathirajan ldquoSpec-trophotometric determination of isoniazid with sodium 12-naphthoquinone-4-sulphonate and cetyltrimethyl ammoniumbromiderdquo Talanta vol 43 no 7 pp 1075ndash1080 1996

[50] M S Kamel ldquoSpectrophotometric determination of isoniazidin pure form and pharmaceutical preparationrdquo World Journalof Chemistry vol 3 no 1 pp 11ndash16 2008

[51] F M A Rind M Y Khuhawar K F Almani and A DRajpar ldquoSpectrophotometric determination of isoniazid indosage forms by derivatizationrdquo Pakistan Journal of AnalyticalChemistry vol 6 no 2 pp 84ndash88 2005

[52] Q-M Li and Z-J Yang ldquoSpectrophotometric study of isoni-azid by using 12-naphthoquinone-4-sulfonic acid sodium asthe chemical derivative chromogenic reagentrdquo Journal of theChinese Chemical Society vol 53 no 2 pp 383ndash389 2006

[53] M Y Khuhawar F M A Rind and K F Almani ldquoSpectropho-tometric determination of isoniazid using 6-methyl-2-pyridinecarboxaldehyde as a derivatizing reagentrdquo Journal of ChemicalSociety of Pakistan vol 20 no 4 pp 260ndash263 1998

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

ISRN Analytical Chemistry 11

[54] J T Stewart and D A Settle ldquoColorimetric determinationof isoniazid with 9 chloroacridinerdquo Journal of PharmaceuticalSciences vol 64 no 8 pp 1403ndash1405 1975

[55] G K Naidu K Suvardhan K S Kumar D Rekha B S Sastryand P Chiranjeevi ldquoSimple sensitive spectrophotometric deter-mination of isoniazid and ritodrine hydrochloriderdquo Journal ofAnalytical Chemistry vol 60 no 9 pp 822ndash827 2005

[56] P Nagaraja K Sunitha R Vasantha and H Yathirajan ldquoNovelmethod for the spectrophotometric determination of isoniazidand ritodrine hydrochloriderdquo Turkish Journal of Chemistry vol26 no 5 pp 743ndash750 2002

[57] B G GowdaM BMelwanki K C Ramesh and J KeshavayyaldquoSpectrophotometric determination of isoniazid in pure andpharmaceutical formulationsrdquo Indian Journal of PharmaceuticalSciences vol 65 no 1 pp 86ndash90 2005

[58] B G Gowda M B Melwanki J Seetharamappa and KC Srinivasa Murthy ldquoSpectrophotometric determination ofisoniazid in pure and pharmaceutical formulationsrdquo AnalyticalSciences vol 18 no 7 pp 839ndash841 2002

[59] S B Kalia G Kaushal and B C Verma ldquoSpectrophotometricmethod for the determination of isoniazidrdquo Journal of the IndianChemical Society vol 83 no 1 pp 83ndash84 2006

[60] A Safavi M A Karimi M R Hormozi Nezhad R Kamaliand N Saghir ldquoSensitive indirect spectrophotometric deter-mination of isoniazidrdquo Spectrochimica Acta A Molecular andBiomolecular Spectroscopy vol 60 no 4 pp 765ndash769 2004

[61] M S Amer Z El-Sherif andMM Amer ldquoSpectrophotometricdetermination of isoniazid nalidixic acid and flumequinethrough ternary complex-formation with Cd(II) and rose Ben-galrdquoEgyptian Journal of Pharmaceutical Sciences vol 35 no 1ndash6pp 627ndash642 1994

[62] M M A Diab B N Barsoum M S Kamel and S Z El-Khateeb ldquoSpectrophotometric determination of isoniazid andrifampicinrdquo Bulletin of Faculty of Pharmacy (Assuit Univ) vol46 no 3 pp 103ndash113 2008

[63] N B Barsoum M S Kamel and M M A Diab ldquoSpec-trophotometric determination of isoniazid and rifampicin frompharmaceutical preparations and biological fluidsrdquo ResearchJournal of Agricultural and Biological Sciences vol 4 no 5 pp471ndash484 2008

[64] H Zhang L Wu Q Li and X Du ldquoDetermination of iso-niazid among pharmaceutical samples and the patientsrsquo salivasamples by using potassium ferricyanide as spectroscopic probereagentrdquoAnalytica ChimicaActa vol 628 no 1 pp 67ndash72 2008

[65] A Safavi and M Bagheri ldquoDesign of an optical sensor forindirect determination of isoniazidrdquo Spectrochimica Acta A vol70 no 4 pp 735ndash739 2008

[66] K Basavaiah and H C Prameela ldquoSpectrophotometric deter-mination of diclofenac sodium using Folin-Ciocalteu reagentin bulk drug and in dosage formrdquo Eastern Pharmacist vol 1pp 61ndash63 2002

[67] T K Murthy G D Sankar and Y S Rao ldquoVisible spectropho-tometric methods for the determination oftrimetazidine dihy-drochloride in pharmaceutical formulationsrdquo IndianDrugs vol39 no 4 pp 230ndash233 2002

[68] K Basavaiah and H C Prameela ldquoSimple spectrophotometricdetermination of acyclovir in bulk drug and formulationsrdquo ILFarmaco vol 57 no 6 pp 443ndash449 2002

[69] C S P Sastry and J S V M Lingeswara Rao ldquoSpectropho-tometric methods for the determination of methotrexate inpharmaceutical formulationsrdquo Analytical Letters vol 29 no 10pp 1763ndash1778 1996

[70] C S P Sastry P Y Naidu and S S N Murty ldquoSpectrophoto-metric methods for the determination of omeprazole in bulkform and pharmaceutical formulationsrdquo Talanta vol 44 no 7pp 1211ndash1217 1997

[71] C S P Sastry A Sailaja T T Rao and D M Krishna ldquoThreesimple spectrophotometric methods for the determination ofsulphinpyrazonerdquo Talanta vol 39 no 6 pp 709ndash713 1992

[72] S Raghuveer A B Avadhanulu and A R Pantulu ldquoSpec-trophotometric determination of gliclazide in its pharma-ceutical dosage forms using Folin-Ciocalteu reagentrdquo EasternPharmacist vol 35 no 1 pp 129ndash130 1992

[73] O Folin and V Ciocalteu ldquoOn tyrosine and tryptophanedeterminations in proteinsrdquo Journal of Biological Chemistry vol73 no 1 pp 627ndash650 1927

[74] AV Prasad P ADevi C S P Sastry andUV Prasad ldquoAssay ofminocycline and tylosin using Folin-Ciocalteu reagentrdquoEasternPharmacist vol 2 no 2 pp 67ndash68 2003

[75] International Conference on Harmonization of technicalrequirements for registration of pharmaceuticals for humanuse ICH harmonized tripartite guideline validation of analyti-cal procedures text andmethodology Q2 (R 1) complementaryguideline on methodology dated 06 November 1996 LondonUK November 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of