“SPECTROSCOPIC METHOD DEVELOPMENT FOR LOSARTON

POTASSIUM IN TABLET”

A project report in Pharmaceutical Analysis By

Ninad .M.Phadke

Under the guidance of :Mrs. Dabhade M.P.

(M.PHARM)

S.N.D. COLLEGE OF PHARMACY,YEOLA

INTRODUCTION TO ANALYTICAL CHEMISTRY “Analytical chemistry deals with methods for determining

the chemical composition of samples of matter.” The drug or dosage form for human use must have

excellent quality and purity, free from any impurity, because these dosage forms directly affects the life, hence their analysis is important which is carried out using analytical methods.

The analytical method development for new API and dosage form involves development and validation of method for testing those API and dosage form in QC laboratories routinely as part of regular Quality control sample testing using the methods of Analytical Chemistry.

Analytical Chemistry involves two branches Qualitative and Quantitative.

BRANCHES OF ANALYTICAL CHEMISTRY

QUALITATIVE ANALYSIS : Qualitative analysis deals with the identification of elements, ions or compounds present in a sample.

QUANTITATIVE ANALYSIS : Quantitative analysis deals with the determinations of how much of one or more constituents are present.

Classification of Analytical Methods

A) CLASSICAL METHODS: For classical qualitative analysis the separated compounds were treated with

reagents that could be recognized by either color, by their boiling or melting points, their

solubility in a series of solvents or their optical activities.

Advantages of Classical Methods:

• Procedure is simple and accurate.• The equipment needed is cheap.• Methods are based on absolute measurements.• Specialized training is not required.

Limitations of Classical Methods:

• Chemical environment is critical.• There is a lack of versatility.• There is a lack of specificity.• Accuracy decreases with decreasing amount.

B) INSTRUMENTAL METHODS :

Measurements of physical properties of analyte such as conductivity, electrode potential, light absorption or emission, mass to charge ratio and fluorescence forms the basis of Instrumental Qualitative Analysis .

Advantages of Instrumental Methods:

Small Samples can be used. High sensitivity is obtained. Measurements obtained are reliable. The determination is very fast. Complex samples can be handled.

Limitations of Instrumental Methods:

Skilled person is required. The sensitivity and accuracy depends on the instruments or wet

chemical methods. Cost of equipment is high. Sizable space is required.

APPLICATIONS OF ANALYTICAL CHEMISTRY

In medicine, analytical chemistry is the basis for clinical laboratory tests, which helps physicians, in diagnosis of various diseases and better recovery of patients.

In industry, it provides the means of testing raw materials and for assuring quality of finished products, like fuel, paints, and other pharmaceuticals products .

Suspected environmental contaminant testing is great achievement by analytical chemistry.

PRINIPLE OF UV VISIBLE SPECTROSOPY

The ultraviolet-visible spectroscopy is a common analytical technique for qualitative and quantitative analysis of solid, liquid or gas samples

The intensity of the absorbance is detected by Beer-Lambert’s law.

Lambert’s Law: At a given concentration (C) of a homogenous absorbing system the intensity of transmitted light decreases exponentially with increasing path length.

-dI / dT = KI Beer’s Law: For a layer of defined path length the intensity of

transmitted light decreases exponentially with increase in concentration (C) of a homogenous absorbing system.

It = Io e –ac

The combination of these laws gives the Beer-Lambert Law,

Log Io/ It = A = abc

Where, A = Absorbance of the solution at a particular

wavelength. Io = Intensity of the light beam.

It = Intensity of beam after passing through solution.

a = Absorptivity of molecule at the wavelength of beam. b = Path length of cell in cm. c = Concentration of solution in gm/lit.

CHOICE OF SOLVENT

SolventMinimum wavelength

( nm )

Acetonitrile 190

Water 190

Cyclohexane 195

Hexane 200

Methanol 200

Ethanol 200

Ether 215

Methylene chloride 220

chloroform 240

Carbon tetrachloride 257

Methods Of Quantitative Spectrophotometric Assay of Medicinal Substances

The concentration of a single component in a sample may be determined by a simple Spectrophotometric methods of absorbances, provided that the other components have a sufficiently small absorbance at the wavelength of measurements.

The single component & multicomponent mixture containing the API can be analyzed by either of following method :

Simultaneous equation method. Multicomponent analysis. Absorption ratio method.

The assay of an absorbing substance may be quickly carried out by preparing solution in a transparent solvent and measuring its absorbance at a suitable wavelength {i.e.maximum absorption (λmax)}.

INSTRUMENTATION OF U.V.

Diagram of double beam UV Spectrophotometer

An instrument for measuring the absorption of UV or visible radiation contains following important components :

Source of light –Hydrogen or deuterium lamp, tungsten lamp.

Prism or Gratings monochromator.Sample container-Quartz cuvettes.DetectorsSignal processor and readout.

Applications of UV-Visible SpectroscopyIdentification of various organic, inorganic

molecules and ions by matching their spectrum with reference spectra.

For qualitative and quantitative analysis of drugs in pharmaceutical industry.

Monitoring of reaction rates (chemical kinetics)Enzyme assaysEnvironmental remote sensingUsed as detectors in various systems like HPLC,

electrophoresis etc.

STEP BY STEP PROCEDURE FOR UV –VISIBLE SPECTROSCOPIC ANALYSIS OF DRUG

Literature Review Objective Of Work Plan Of Work Experimental Work Results and discussionMethod Validation Conclusion

PLAN OF WORKLosarton Potassium

Calibration Of Apparatus

Calibration Of UV Instrument

Selection of suitable solvent

Determination of max

Calibration curve

Estimation of drug from tablet formulation

Recovery studies

Result and conclusion

OBJECTIVE OF WORK

The objective of this investigation was to devise simple, precise, rapid and economical method for the estimation of Losarton Potassium in bulk drug and the tablet formulation.

Tablets procured from the market were analyzed by the proposed method. In this method, the tablets were crushed and dissolved in solvent and diluted further. Sufficient amounts of the samples were withdrawn and their absorbances were noted at 238.4 nm against reagent blank.

Optical characteristics and precision

Absorption

maxima(nm)

238.4nm

Absorption maxima(nm) 238.4nm

Beer’s law limit (g/ml) 03-30µg/ml

Correlation coefficient(r2) 0.9871

Regression equation 0.0154x + 0.0198

TABLET ANALYSISSR. NO. Amt taken in

(g/ml)

Amt found in

(g/ml)

Amt found in

%

 

1. 18 17.99 99.94

2. 18 18.01 100.05

3. 24 23.99 99.95

4. 24 24.01 100.04

5. 30 29.99 99.96

6. 30 30.01 100.03

Statistical validation of tablet formulation

Amt

found in

(%)

MEAN Standard

deviation

Coeffecie

nt of

variation

Standard

error

 

100.03 23.83 ± 0.15379 0.932 0.002124

Recovery study of Losarton PotassiumName of drug

Level

of

Perce

ntag

e

Reco

very

Conc

.

used

µg/

ml

Conc

.

adde

d

µg/

ml

Total

conc.

absor

banc

e

Conc.

Recov

ered in

µg/ml

%

Recov

ery

MEA

N

 

Losarton

Potassium

 

50 12 6 18 0.298 17.99 99.94 99.97

 

50 12 6 18 0.297 18.01 100.05

50 12 6 18 0.298 17.99 99.94

100 12 12 24 0.440 23.99 99.95 99.99

100 12 12 24 0.441 24.01 100.04

100 12 12 24 0.440 23.99 100.00

150 12 18 30 0.597 29.99 99.96 99.98

150 12 18 30 0.596 30.01 100.03

150 12 18 30 0.597 29.99 99.96

Statistical validation of recovery studies of tablet formulation

Level of

recovery

MEAN %

Recovery

Standard

Deviation

Coeffecien

t Of

Variation

Standard

Error

 

50 % 99.97

 

±0.076895 0.466 0.001062

100 % 99.99 ± 0.15379 0.932 0.002124

150 % 99.98 ±0.30758 1.864 0.004248

Statistical Results Of Analysis

Sr. No. Tablet Label

claim(mg)

S.D.* %Recover

y*

1. LOSAR 50 ±0.15379 99.98 %

Thus the statistical study was performed.The value obtained for S.D seemed to be within range. This showed the suitability of procedure.

METHOD VALIDATION

Linearity-Linearity was observed in the range of 03-30g/ml. The calibration curve yielded coefficient of correlation (r2) 0.9871.

Assay result- Losarton Potassium tablets of a marketed brand was analyzed by proposed methods, the percentage in tablet were determined and presented in the above table.

Accuracy and precision-The % Recovery was found to be in the range of 99.80-100.5%,indicates the non-interference from the formulation excipient and confirm the accuracy and precision of the method.

CONCLUSION

All the above result indicates that, the method employed here is a very simple, economical and can be used for routine analyses of the drug, Losarton Potassium.

The proposed method was found to be accurate, simple and rapid for routine analysis of the drug. The recovery was 100.43% (LOSAR) which is close to 100% indicating reproducibility and accuracy of the method.

References Govt. of India, Ministry of Health and Family Welfare. Vol. 2. Delhi:

Publication by Controller of Publication; 2007. Indian Pharmacopoeia;

pp. 484–554.

British Pharmacopoeia. (International ed.) 1993;Vol. 1:429, 483.

Published on the Recommendation of the Medicines Commissions

Pursuant to Medicines Act 1968, 1993.

 Martindale, The Extra Pharmacopoeia, 33rd ed., Royal Pharmaceutical

Society, London, 2002; pp 921-922.

 United States Pharmacopoeia 29 NF 24, Published on the

Recommendation of the Medicines Commissions Pursuant to Medicines,

page no. 587 

Skoog, West, Holler, Crouch, “Fundamentals of analytical chemistry” ,

eighth edition, 2009 (Indian edition), cengage learning India pvt ltd ,

New delhi, pageno. 271-280.

 A.V Kasture, K.R Mahadik, S.G Wadodkar, H.N. More, “A textbook of

pharmaceutical analysis, Instrumental methods” , Nirali Prakashan,

vol.2, 9th edition, page no. 5-7, 28-30

THANKING YOU