http://www.ijapbr.com/
International journal of Applied Pharmaceutical and Biological Research, 2017; 2(2):11-27
Research Article ISSN : 2456-0189
11
STABILITY INDICATING NEW RP-HPLC METHOD FOR THE
DETERMINATION OF ROSUVASTATIN CALCIUM IN PURE AND TABLETS
DOSAGE FORMS
Mohamed El-Kassem M Hassouna*, Hafsa Omar Salem
Chemistry Department, Faculty of Science 62514, Beni-Suef University, Beni-Suef , Egypt
ABSTRACT
A new, specific, precise, simple, and accurate RP-HPLC method is developed and validated for the determination of Rosuvastatin calcium (ROS-Ca) in pure and tablets dosage forms. The method is performed on the Agilent Eclipse XDB C8 column (250 mm X 4.6 mm, 5μm particle size, using buffer solution of pH 4.5 containing 0.05M sodium dihydrogen phosphate: acetonitrile (50:50 v/v) as the mobile phase at a flow rate of 1.2 mL/min, injection volume 10 µL and UV detection at 245 nm. The total run time is 5.0 min. Linear relationship are obtained in the range 5-100 µg/mL and Rt values of 3.684 min for ROS-Ca with correlation coefficient (r) = 0.9995, limit of detection 1.50 µgmL-1 and limit of quantitation is 4.56 µgmL-1 for ROS-Ca. The overall recovery is 100 ± 2 % ; the relative standard deviation for precision and intraday precision is less than 2.0 %. Also, the forced degradation studies as acidity, alkalinity, oxidation, heat and photo degradation are performed according to ICH guidelines. The method is validated according to ICH guidelines and USP requirements for new methods, which include accuracy, precision, specificity, LOD, LOQ, robustness, ruggedness, linearity and range. Hence this RP-HPLC method is suitable for quality control of raw materials and finished products.
Key words: Rosuvastatin Calcium; Stability; Assay; tablets dosage form; ICH; USP; HPLC
INTRODUCTION
Rosuvastatin Calcium (ROS-Ca); is chemically known as [(3R,5S,6E)-7-[4-(4-fluorophenyl)-6-(1-
methyleythyl) -2- [methyl (methyl sulfonyl) amino] pyrimidine-5- yl]-3,5-dihydroxyhept-6-enoate]. It has a
molecular formula of C44H45CaF2N6O12S2 and a molecular weight of 1001 [1, 2]. (Fig 1).
Rosuvastatin Calcium is a white or almost white, hygroscopic powder, slightly soluble in water, freely soluble in methylene chloride, practically insoluble in anhydrous ethanol. Store in airtight container, protected from
light, at temperature of 2°C to 8°C.[1,2].
Rosuvastatin is available in tablet form containing 5 mg, 10 mg, 20 mg and 40 mg of the active ingredient,
Rosuvastatin calcium. The excipients are: microcrystalline cellulose NF, lactose monohydrate NF, tribasic
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
12
calcium phosphate NF, crospovidone NF, magnesium stearate NF, hypromellose NF, triacetin NF, titanium
dioxide USP, yellow ferric oxide, and red ferric oxide NF. [3].
Figure 1: Chemical structures of Rosuvastatin Calcium.
Rosuvastatin selectively and competitively inhibits HMG-CoA reductase, the rate-limiting enzyme that
converts HMG-CoA to mevalonate, a precursor of cholesterol. Rosuvastatin has been shown to have high
uptake and selectivity in the liver, the target organ for cholesterol lowering. Rosuvastatin produces its lipid-modifying effects in two ways: it increases the number of hepatic LDL receptors on the cell-surface to
enhance the uptake and catabolism of LDL, and it inhibits hepatic synthesis of VLDL, which reduces the total
number of VLDL and LDL particles [3].
Literature review revealed that various analytical methods have been described for the determination of
Rosuvastatin Calcium including spectrophotometric [4-15], thin layer chromatography (TLC) [16], capillary
electrophoresis [17],mass spectrometry [18-24],liquid chromatography (HPLC, UPLC)[25–50],
electrochemical methods [51,52] and complexometric titration [53] have been developed for the estimation
of ROS-Ca in pure or in dosage forms.
The aim of the present work is to develop a new, simple, sensitive, short retention time and accurate RP-HPLC
method for the determination of ROS-Ca in pure or in dosage forms and application to assay dosage forms
with high sensitivity, selectivity that are required to be in routine quality control analysis and validate the
developed methods according to ICH guidelines [54].
MATERIALS AND METHOD
Chemicals and Reagents
Acetonitrile, Methanol HPLC-grade, Sodium dihydrogen phosphate monohydrate analytical grade,
Triethylamine HPLC grade, Hydrochloric Acid analytical grade, Sodium Hydroxide reagent grade and distilled Water are procured from (scharlau, Spain).
Pure samples
Pure samples of Rosuvastatin Calcium were kindly supplied by SPIMACO Pharmaceutical Company,
Alexandria, Egypt with claimed purity of 100.5%. The content of Rosuvastatin Calcium in BP Pharmacopeia is
in the range 97.0 to 102.0 percent.
Pharmaceutical dosage form
Six pharmaceutical preparations viz., Cholerose 10 mg Tab, Crestore 10 mg Tab, Estero-map 10 mg Tab, Advochol 10 mg Tab, Justechol 10 mg Tab and Crestolip 10 mg Tab are purchased from the local Egyptian
market. Each tablet is claimed to contain 10.4 mg of Rosuvastatin Calcium.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
13
Mobile phase preparation: Buffer: Acetonitrile (50:50)
Sodium dihydrogen buffer was prepared by dissolving 6.85 gm of sodium dihydrogen phosphate
monohydrate in 700 mL distilled water and sonicated to dissolve, adjust pH to 4.5 by orthophosphoric acid
solution or triethylamine. Make up to 1000 mL with distilled water, filter and degass mixtures of buffer and
acetonitrile (50:50) through 0.45μ membrane filter under vacuum pump.
Diluent:
Methanol HPLC-grade
Apparatus
HPLC system (Shimadzu LC SPD 20 A) with a detector (dual wavelength), equipped with a binary pump, Autosampler, oven CTO-20A/20AC with temperature range (10-85◦C), LC Solution software.
Mettler pH Meter.
Shimadzu analytical balance.
Ultra-sonic bath.
HPLC Chromatographic Conditions
Chromatographic separation is performed on column Agilent Eclipse XDB- C8 (250 X 4.6 mm i.d, 5 µm particle
size) .Using a mobile phase mixture of sodium dihydrogenphosphate buffer and acetonitrile in the ratio of
50:50 % v/v at ambient temperature, flow rate of 1.2 mL/min, UV detection is performed at 245 nm, injection volume is 10 μL and run time is 5.0 min.
Preparation of standard and samples solution
Stock solutions of Rosuvastatin Calcium (1000 μg /mL)
100mg of ROS-Ca of the working standard are weighed accurately, transferred into 100 mL volumetric flask,
add 70 mL of diluent and sonicate to dissolve. Complete the volume to the mark with the same diluent and
mix well.
Working standard solutions of Rosuvastatin Calcium (50 μg /mL)
Transfer 5 mL from the stock solution of ROS-Ca into 100 mL volumetric flask, add 70 mL of the diluent and sonicate to dissolve, complete to the mark with the same diluent and mix well. The obtained chromatogram is
shown in Figure 2.
Figure 2: HPLC chromatogram of standard solution of (50μg/mL) of Rosuvastatin Calcium.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
14
Application to pharmaceutical formulations
Weigh not less than 10 tablets from products mentioned above and determine the average weight of one
tablet. Grind to fine powder. Transfer an accurately weighed weight from powdered tablets equivalent to the
average weight of one tablet into 200-mL volumetric flask. Add about 100 mL diluent, sonicate till dissolve
and complete to volume with the same diluent and mix well. Filter through 0.45 μm syringe filter and inject into the chromatographic system. The obtained chromatogram is shown in (Figure 3). Also, the standard
addition technique has been carried out to assess the validity of the method by spiking the pharmaceutical
formulation with known amount of standard solution of ROS-Ca. The recovery of the added standards is then
calculated after applying the proposed method.
Construction of calibration curves:
Different concentrations of ROS-Ca equivalent to (5–100) μg /mL, are separately withdrawn from their
respective stock standards into separate series of 100 mL volumetric flasks, and the volumes are made up to
the mark with the diluent. Duplicate 10 µL injections are made for each concentration maintaining the flow
rate at 1.2 mL/min and the effluent is UV- scanned at 245 nm. The chromatographic separation is performed
following the procedure under chromatographic conditions. The chromatograms are recorded, peak areas of
ROS-Ca are determined and the calibration curves relating the obtained integrated peak areas to the
corresponding concentrations are constructed and the regression equations are performed
. RESULTS AND DISCUSSION
The goal of this study is to develop HPLC assay for the analysis of Ros.Ca drug in its pure form and in
pharmaceutical formulation. Although, the literature is full of a number of HPLC methods for the
determination of rosuvastatin calcium [43-50] due to the high accuracy and precision obtained by HPLC
technique, the present method is devoted, also, for the same goal with aim of achieving better results under
much simpler conditions. Chromatograms are obtained with Rt value of 3.684 min for ROS-Ca with
correlation coefficients (r) =0.9995, limit of detection 1.50 µg mL-1 and limit of quantitation is 4.56 µg mL-1 for
ROS Ca. No occurrence of interfering peaks.
Methods development and optimization
Different developing systems of different compositions and ratios are tried including: methanol: water
(50:50, v/v), acetonitrile: water (50:50, v/v), but no peak was observed for ROS-Ca. Different flow rates are tried (0.7, 1.0, 1.2 and 1.5 mL/min), scanning wavelengths (200 -400 nm) are also tried. Preliminary studies
involved trying C18, C8 reversed-phase columns. The best developing system proved to be sodium dihydrogen phosphate buffer pH (4.5): ACN (50:50, v/v) at flow rate of 1.2mL/min and at wavelength of 245.0
nm using column Agilent Eclipse XDB- C8 (250 mm X 4.6 mm i.d., 5μm). This selected developing system
allows good separation and sharp peak with good Rt values without tailing of the separated bands and good
theoretical plates.
Validation of the Analytical Method
The method is validated, in accordance with ICH guidelines (ICH Q2R1), for system suitability, precision,
accuracy, linearity, specificity, ruggedness, robustness, LOD and LOQ [54].
Linearity and range
The linearity of the proposed methods is obtained in the concentration range (5.0 -100.0 μg/mL) for
Rosuvastatin Calcium. Calibration curves are composed by plotting peak areas against the corresponding
concentrations. The obtained coefficients of regression are 0.9995 for ROS-Ca. Linearity results are shown in
Table 1.
Repeatability
Repeatability of the method is evaluated by calculating the RSD of the peak areas of six replicate injections for
the standard concentration (50.0 μg/mL) of ROS-Ca. Results are examined as % RSD values of the
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
15
concentrations of the determined drug. Low values of % RSD (less than 2) indicate high precision of the
method as shown in Table 1.
Table 1: Regression and validation parameters of the proposed HPLC method for determination of
ROS-Ca
ROS-Ca Parameter
Linear
5-100 range (µg/mL) 32348.6829 Slope 8420.0893 Intercept
0.9995 Correlation coefficient 1.50 LOD a (µg/mL) 4.56 LOQ a (µg/mL)
0.013 Repeatability b
aLimit of detection (3.3× σ /Slope) and limit of quantitation (10× σ /Slope). b Repeatability for n≥5, RSD ≤2.
Table 2: Data of Accuracy for ROS-Ca.
ROS-Ca. Standard (μg/mL) ROS-Ca
μg/mL (Injected) μg/mL (found) Recovery%
25 25 25.37 101.48%
25 25.33 101.33%
25 25.30 101.20%
50 50 50.03 100.06%
50 50.04 100.09%
50 50.07 100.13%
80 80 79.39 99.24%
80 78.78 98.47%
80 78.64 98.30%
Accuracy (Mean) 100.04
Table 3: Determination of ROS-Ca in pharmaceutical formulation by the proposed HPLC method and
application of standard addition
technique
Recovery % Added(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca
99.13 10 Crestolip 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
100.31 20 98.53 30
99.32±0.91
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
16
Table 4: Assay result for the determination of Rosuvastatin Calcium in pharmaceutical formulation by
the proposed HPLC method.
limit % Recovery % Conc.(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca ROS-Ca
(90 -110) 104.75% 50 Cholerose 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
104.72% 104.52% 104.81% 104.87% 104.92%
104.76±0.13
Table 5: Assay result for the determination of Rosuvastatin Calcium in pharmaceutical formulation by
the proposed HPLC method.
limit % Recovery % Conc.(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca ROS-Ca
(90 -110) 105.30% 50 Crestore 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
106.69% 106.39% 107.76% 107.26% 103.43%
106.14±1.47
Table 6: Assay result for the determination of Rosuvastatin Calcium in pharmaceutical formulation by
the proposed HPLC method.
limit % Recovery % Conc.(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca ROS-Ca
(90 -110) 105.48% 50 Estero-map 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
105.29% 105.56% 105.61% 105.55% 105.86%
105.55±0.17
Table 7: Assay result for the determination of Rosuvastatin Calcium in pharmaceutical formulation by
the proposed HPLC method.
limit % Recovery % Conc.(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca ROS-Ca
(90 -110) 100.52% 50 Advochol 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
100.58% 100.32% 100.48% 100.41% 100.55%
100.47±0.09
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
17
Table 8: Assay result for the determination of Rosuvastatin Calcium in pharmaceutical formulation by
the proposed HPLC method.
limit % Recovery % Conc.(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca ROS-Ca
(90 -110) 104.27% 50 Justechol 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
104.31% 104.29% 104.75% 104.30% 104.31%
104.37±0.18
Detection and Quantitation limits
These approaches are based on the Standard Deviation of the Response and the Slope. A specific calibration curve should be studied using samples containing an analyte in the range of LOD and LOQ. The residual
standard deviation of a regression line or the standard deviation of y-intercepts of regression lines may be
used as the standard deviation. LOD= 3.3 × σ /slope and LOQ =10 × σ /slope, where σ = the standard
deviation of the response Table 1.
Accuracy and recovery
Accuracy of the proposed method is calculated as the percentage recoveries of pure samples of the studied
drugs. Accuracy is assessed using three different concentrations (25, 50 & 80 μg/mL) for ROS-Ca within the
linearity range (i.e. three concentrations and three replicates). Concentrations are calculated from the
corresponding regression equations. The mean % recoveries for ROS-Ca are between 98.0 % to 102 %. These data are shown in Table 2. Accuracy is further assessed by applying the standard addition technique to
Crestolip® 10 mg Tab, where good recoveries are obtained revealing that there is no interference from
excipients, Table 3.
Formulation assay
The validated method is applied to the determination of ROS-Ca in commercially available pharmaceutical
preparations available in the local Egyptian market. The results of the assay indicate that the method is
selective for the analysis of all mentioned products without interference from the excipients. The results are
displayed in Tables 4 - 9.
Intermediate precision (ruggedness)
Intermediate precision expresses within-laboratories variations: different days, different analysts, different
equipment's, etc. Good results are obtained and presented in Table10.
Robustness
The robustness of the proposed method is evaluated in the development phase where the effects of different
factors on the method are studied to obtain the optimum parameters for complete separation. Robustness of
the method is studied by deliberately varying parameters like flow rate (±0.1 mL/min) and studying the
effect of changing mobile phase pH by (± 0.2), acetonitrile composition (±5%) and column temperature
changed (±5°c). The low values of the % RSD, as given in Table11, indicate the robustness of the proposed
method.
System suitability
System suitability testing is an integral part of many analytical procedures. The tests are based on the concept
that the equipment, electronics, analytical operations and samples to be analyzed constitute an integral
system that can be evaluated as such. System suitability is checked by calculating tailing factor (T), column
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
18
efficiency (N), resolution (Rs) factors. All calculated parameters are within the acceptable limits indicating
good selectivity of the method and ensuring system performance, Table 12.
Stability of the analytical solution
To demonstrate the stability of standard solution during analysis, solution is analyzed over a period of 24 h at room temperature and refrigerator. The results showed that for all the solutions, the retention times and
peak areas of ROS-Ca remained almost unchanged (RSD<2.0%) indicating that no significant degradation
occurred within this period, i.e. solutions are stable for at least 24 h, which is sufficient to complete the whole
analytical process. The results are displayed in Table 13.
Specificity
Placebo interference:
Method specificity is determined for samples of studied compounds and placebo matrix containing all
excipients present in the finished product. No interferences are detected at the retention time of ROS-Ca. Also,
the inactive ingredients content of the pharmaceutical preparations do not interfere with HPLC procedure.
Forced degradation:
Forced degradation of the active pharmaceutical ingredient (API) was carried out according to ICH guidelines
(ICH, Q2B) in acid, base, oxidation, photo and heat.
Table 9: Assay result for the determination of Rosuvastatin Calcium in pharmaceutical formulation by
the proposed HPLC method.
limit % Recovery % Conc.(µg/mL) Pharmaceutical formulation
ROS-Ca ROS-Ca ROS-Ca
(90 -110) 105.53% 50 Crestolip 10 mg Tab
ROS Ca, 10.4 mg(claimed)
Mean ± RSD
105.47% 105.59% 105.80% 105.53% 105.53%
105.57±0.11
Table 10: Ruggedness of the method
Table 11: Robustness of the method
Parameter(%RSD) ROS-Ca
Flow rate change (±0.1 mL/min) 1.253
pH change of mobile phase (±0.2) 1.425
Wave length change (±0.2nm) 0.889
Column temperature change(±5ₒC) 0.643
Parameter(%RSD) ROS-Ca
Intraday 0.159
Interday 0.989
Analyst to Analyst 0.168
Column to Column 0.353
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
19
Table 12: System suitability testing parameters of the developed method
Item Obtained Value Reference values
ROS-Ca
Tailing factor 1.110 T ≤ 2
Resolution - Rs>2
Selectivity 3.5 k’ > 2
Injection precision 0.178 RSD ≤1%
Retention time (Rt) 0.122 RSD ≤1%
Number of theoretical plates(N) 5530.772 N > 2000
Table 13: Result of stability of analytical solution
Table 14: Results of analysis for forced degradation study samples using the proposed method,
indicating percentage degradation of
ROS-Ca
Parameter Rosuvastatin Calcium
Effect Observed tR Peak Area Degradation %
Test Without Effect(control) 3.684 1668239 -
Oxidation Effect 3.705 220023 86.81
Alkali Effect 3.198 1550541 7.05
Acid Effect 3.857 1494943 10.38
Light Effect (Sun light) 3.687 1621340 2.81
Heat Effect 3.682 1656890 0.68
Placebo No peak observed No area observed -
Photo degradation:
The powder of working standard ROS-Ca is kept under sunlight for 48 hours. From this powder, accurately
weigh 10.0 mg and transfer to 200-mL volumetric flask. Add 100 mL of diluent and sonicate to dissolve. Make
up to the mark with water and mix well. Filter through 0.45 μm membrane filter, reject the first portion then
analyze by HPLC. No interference is found at the retention time of ROS-Ca and no degradation products
appeared after light degradation.
Heat degradation:
Keep the powder sample of the working standard ROS-Ca in dry oven at 80ºC for 6 hours. Similarly, accurately weigh 10.0 mg and transfer to 200-mL volumetric flask. Add 100 mL of diluent and sonicate to
dissolve. Make up to the mark with water and mix well. Filter through 0.45 μm membrane filter, reject the
first portion then analyze by HPLC. No interference is found at the retention time of ROS-Ca and no
degradation products appeared after heat degradation.
Condition ROS-Ca
Fridge (2-8°C) 100.69%
Room temperature (25°C) 100.67%
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
20
Acid degradation:
Accurately weigh 10.0 mg of the working standard ROS-Ca powder and transfer into 200-mL conical flask.
Add 100 mL of diluent and sonicate to dissolve, add 10 mL of 0.1 N HCl then keep the acidified solutions in
room temperature for one day. Then complete to the mark with diluent and mix well. Filter and analyze by
HPLC. No interference is found at the retention time of ROS-Ca and no degradation products appeared after
acid degradation.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
21
Figure 3.Typical HPLC chromatogram of sample solution (50μg/mL) of each (a)Cholerose 10 mg Tab,
(b)Crestore 10 mg Tab, (c)Estero-map 10 mg Tab, (d) Advochol 10 mg Tab, (e) Justechol 10 mg Tab
and (f) Crestolip 10 mg Tab.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
22
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
23
Figure 4: HPLC chromatograms of 50 µg/ml solution of (a) Rosuvastatin Calcium after exposure to (b)
heat degradation, (c) photo degradation, (d) acid degradation, (e) base degradation and (f) oxidative
degradation.
Base degradation:
Similarly, weigh 10.0 mg of the working standard ROS-Ca powder and transfer it into 200-mL conical flask.
Add 100 mL of diluent and sonicate to dissolve, add 10 mL of 0.1 N NaOH then keep the alkaline solutions in
room temperature for one day. Then complete to the mark with diluent and mix well. Filter and analyze by
HPLC. No interference is found at the retention time of ROS-Ca but its peak area is affected after base
degradation. The degradation percent is calculated and displayed in (Table 14).
H2O2 degradation:
Finally, weigh 10.0 mg of the working standard ROS-Ca powder and transfer into 200-mL conical flask. Add
100 mL of diluent and sonicate to dissolve, add 15 mL of 3.0 % H2O2 then keep at room temperature for one
day. Then complete to the mark with the diluent and mix well. Filter and analyze by HPLC. No interference is found at the retention time of ROS-Ca but some degradation products appeared after the H2O2 treatment. The
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
24
degradation percent is calculated and displayed in (Table 14). The corresponding chromatogram obtained is
shown in (Figure 4).
CONCLUSION
The proposed RP-HPLC method for determination of Rosuvastatin Calcium in pure and tablets dosage form is,
precise, specific, accurate and less time consuming for analysis, low cost and rapid. The results of forced
degradation undertaken according to the International Conference on Harmonization (ICH) guidelines reveal
that the method is selective. Based on the above results, the analytical method is valid, fit for use and can be
used for regular routine analysis and stability study.
Acknowledgement
The authors are thankful to chemist Mahmoud A Mohamed, QQC Supervisor at EPCI part of HIKMA group,
Beni-Suef, Egypt for his valuable help and providing training and guidance for the second author throughout
the performance of the present work.
REFERENCES
1) British Pharmacopoeia, Stationary Office, Medicines and Healthcare Products Regulatory Agency, London, Vol. II, 2017.
2) European Pharmacopeia 9th edition 2017.
3) https://www.fda.gov/downloads/Drugs/DevelopmentApprovalProcess/DevelopmentResources/UCM20
5893.
4) Ramadan A, Mandil H, Alshelhawi N. Spectrophotometric Determination of Rosuvastatin Calcium in Pure
Form and Pharmaceutical Formulations by The Oxidation Using Iodine and Formation Triiodide
Complex in Acetonitrile. Int J Pharm Pharm Sci. 2014; 6(5):579-585.
5) Gupta A, Mishra P, Shah K. Simple UV Spectrophotometric Determination of Rosuvastatin Calcium in Pure
Form and in Pharmaceutical Formulations. E-Journal of Chemistry. 2009; 6(1):89-92.
6) Lahare RY,Phuge AN,Gite AL,Jadhav AK. A Review on Ultraviolet Spectrophotometric Determination of RosuvastatinCalcium in Marketed Formulation. Int J Pure App Bio sci. 2014; 2 (6):169-174.
7) Uyar B, Celebier M, Altinoz S. Spectrophotometric determination of rosuvastatin calcium in tablets.
Pharmazie. 2007; 62: 411–413.
8) Pandya CB, Channabasavaraj KP, Shridhara HS. Simultaneous Estimation of Rosuvastatin Calcium and
Ezetimibe in Bulk and Tablet Dosage Form by Simultaneous Equation Method. IJCRGG. 2010; 2(4):2140-
2144.
9) Pandya N, Mashru RC. Simultaneous Estimation of Rosuvastatin Calcium and Clopidogrel Bisulfate in
Combined Pharmaceutical Formulation. IJDFR. 2012; 3(5):40-53.
10) Anuradha GK, Vishal SD. Simultaneous Estimation of Rosuvastatin And Ezetimibe by Ratio Spectra Derivative Spectrophotometry Method in Their Fixed Dosage Forms. IJPRIF. 2010; 2(1):404-410.
11) Savita NM, Promod SH, Chandrashekhar BL. Quantitative Determination of Rosuvastatin Calcium and
Niacin Individually and Combined Tablet Dosage Form by Using UV-VIS Spectrophotometer.
International Journal of Pharma Research & Review. 2015; 4(6):37-43.
12) Solanki C, Patel N, Patel V, Patel D, Vaishy R. Development and Validation of First Order Derivative
Spectrophotometric Method for Simultaneous Estimation of Rosuvastatin Calcium and Aspirin in Capsule
Dosage Form. Der Pharmacia Lettre 2012; 4(3):947-953 .
13) Anuradha GK,Vishal SD. Simultaneous UV spectrophotometric Estimation Of Rosuvastatin And Ezetimibe
In Their Combined Dosage Forms. International Journal of Pharmacy and Pharmaceutical Sciences.
2010; 2(1):131-138.
14) Mandwal PS, Patel PR, Agarwal KM, Surana SJ. Q-Absorbance and Multicomponent UV -
Spectrophotometric Methods for Simultaneous Estimation of Rosuvastatin calcium and
Fenofibrate in Pharmaceutical Formulation.Der Pharmacia Lettre 2012; 4(4):1054-1059.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
25
15) Patel B, Jadav A, Solanki H, Parmar S, Parmar V, Captain A. Development and Validation of Derivative
Spectroscopic Method for the Simultaneous Estimation of Rosuvastatin Calcium and Fenofibrate in
Tablet. International Journal of Pharma Research & Review. 2013; 2(7):1-6.
16) Tank PH, Vadalia KR, Dedania ZR. Development and Validation of Hptlc Method for Simultaneous
Estimation of Rosuvastatin Calcium and Aspirin in Capsule Dosage Form. IJPSR. 2102; 3(10):3867-3870. 17) Su¨slu¨ I, Celebier M, Altıno¨z S. Determination of Rosuvastatin in Pharmaceutical Formulations by
Capillary Zone Electrophoresis. Chromatographia. 2007; 66(1):65-72.
18) Zhang D, Zhang J, Liu X, Wei C, Zhang R, Song H, Yao H. Validated LC-MS/MS Method for the
Determination of Rosuvastatin in Human Plasma: Application to a Bioequivalence Study in Chinese
Volunteers. Pharmacology & Pharmacy. 2011; 2: 341-346.
19) Ishaq BM, Prakash KV, Kumar CH, Rani GU, Mohan GK. Development and validation of LC-MS method for
the determination of Rosuvastatin Hydrochloride in human plasma. J. Chem. Pharm. Res. 2010;
2(6):324-333.
20) Hull CK, Penman AD, Smith CK, Martin PD. Quantification of rosuvastatin in human plasma by automated
solid-phase extraction using tandem mass spectrometric detection. Journal of Chromatography B. 2002;
772(2): 219–228.
21) Gao J, Zhong D, Duan X, Chen X. Liquid chromatography/negative ion electrospray tandem mass
spectrometry method for the quantification of rosuvastatin in human plasma: Application to a
pharmacokinetic study. Journal of Chromatography B 2007; 856(1–2): 35–40. 22) Xu DH, Ruan ZR, Zhou Q, Yuan H , Jiang B. Quantitative determination of rosuvastatin in human plasma by
liquid chromatography with electrospray ionization tandem mass spectrometry. Rapid Commu-nications
in Mass Spectrometry. 2006; 20(16): 2369-2375, doi:10.1002/rcm.2542 .
23) Siddartha B, Babu S. Estimation and Validation for Determination of Rosuvastatin In Human Plasma by
Lc/Ms/Ms Method. JGTPS. 2014; 5(3):1979–1988.
24) Zaid A, Al Ramahi R, Cortesi R, Mousa A, Jaradat N, Ghaza N, Bustami R. Investigation of the
Bioequivalence of Rosuvastatin 20 mg Tablets after a Single Oral Administration in Mediterranean Arabs
Using a Validated LC-MS/MS Method. Sci. Pharm. 2016; 84: 536–546; doi:10.3390/scipharm84030536.
25) Sheth A, Patel KN, Ramlingam B, Shah N. Simultaneous Estimation of Rosuvastatin Calcium and
Clopidogrel Bisulphate From Bulk and Commercial Products Using a Validated Reverse Phase High Performance Liquid Chromatographic Technique. IRJP. 2012; 3(11):154-157.
26) Bhati LK, Kumar MV. Bilayer Tablet of Rosuvastatin Calcium and Fenofibrate: An Assessment Prior To
Formulation Design. Ejpmr. 2016; 3(6): 391-395.
27) Turabi ZM, Khatatbeh OA. Stability-Indicating RP-HPLC Method Development and Validation for the
Determination of Rosuvastatin (Calcium) In Pharmaceutical Dosage Form. Int. J. Pharm. Sci. Drug Res.
2014; 6(2): 154-159 .
28) Substances in solid oral dosage form using the developed and validated, stability indicating, RP-UPLC
method,Chapter-4, "Determination of Related Substance in Rosuvastatin Tablets by RP-UPLC
Method",117-146. 29) Thriveni J, Rambabu R, Rao JV, Vidyadhara S. Development and Validation of Rp-Hplc Method For
Simultaneous Estimation of Rosuvastatin Calcium and Fenofibrate In Bulk and Pharmaceutical Dosage
Forms. IJRPC. 2013; 3(2):208-212.
30) Petkovska R, Cornett C, Dimitrovska A. Development and Validation of Rapid Resolution RP-HPLC Method
for Simultaneous Determination of Atorvastatin and Related Compounds by Use of Chemometrics.
JournalAnalytical Letters 2008; 41( 6):992-1009.
31) Varma PD, Rao AL, Dinda SC. Development and Validation of Stability Indicating Rp-Hplc Method for
Simultaneous Estimation of Rosuvastatin And Ezetimibe in Combined Tablet Dosage Form. RASAYAN J
Chem. 2012; 5(3):269-279.
32) Karunakaran A, Subhash V, Chinthala R, Muthuvijayan J. Simultaneous Estimation of Rosuvastatin
Calcium and Fenofibrate in Bulk and in Tablet Dosage Form by UV-Spectrophotometry and RP-HPLC. S J
Pharm Sci. 2011; 4(1): 58-63.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
26
33) Pandya CB, Channabasavaraj KP, Shridhara HS. Simultaneous Estimation of RosuvastatinCalcium and
Ezetimibe in Bulk and Tablet Dosage Form by Simultaneous EquationMethod. Int J ChemTech Res.
2010; 2(4):2140-2144.
34) Tajane D, Raurale AM, Bharande PD, Mali AN, Gadkari AV, Bhosale VR. Development and validation of a
RP-HPLC-PDA method for simultaneous determination of Rosuvastatin calcium and Amlodipine besylate in pharmaceutical dosage form. J Chem Pharm Res. 2012; 4(5):2789-2794.
35) Swathi Sd, Hemant KT, Vara Prasada RK, Srinivasa RY. Validated Rp-Hplc Method for Simultaneous
Determination of Rosuvastatin Calcium and Ezetimibe in Pharmaceutical Dosage Form. Int J Pharm
Pharm Sci. 2015; 7(4): 209-213.
36) Chakraborty AK, Mishra SR, Sahoo HB. Formulation of Dosage Form of Rosuvastatin Calcium And
Development of Validated Rp-Hplc Method for Its Estimation. International Journal of Analytical and
Bioanalytical Chemistry. 2011; 1(3): 89-101.
37) Sultana N, Arayne MS, Ali SN. An Ultra-Sensitive LC Method for Simultaneous Determination of
Rosuvastatin, Alprazolam and Diclofenac Sodium in API, Pharmaceutical Formulations and Human Serum
by Programming the Detector. J Anal Bioanal Techniques 2012; 3(7):1-6.
38) Sultana N, Arayne MS, Shah SN, Shafi N, Naveed S. Simultaneous Determination of Prazosin, Atorvastatin,
Rosuvastatin and Simvastatin in API, Dosage Formulations and Human Serum by RP-HPLC. Jnl Chinese
Chemical Soc. 2010; 57(6):1286.
39) Solanki C, Patel N. Development and Validation of Rp-Hplc Method for Simultaneous Estimation of Rosuvastatin Calcium and Aspirin in Capsule Dosage Form . Int J Pharm Bio Sci. 2012; 3(3): 577 – 585.
40) Sultana N, Arayne MS, Iftikhar B. Simultaneous Determination of Atenolol, Rosuvastatin, Spironolactone,
Glibenclamide and aproxen Sodium in Pharmaceutical Formulations and Human Plasma by RP-HPLC. Jnl
Chinese Chemical Soc. 2008; 55(5): 1022–1029.
41) Arayne MS, Sultana N,Tabassum A, Ali SN,Naveed S. Simultaneous LC determination of rosuvastatin,
lisinopril, captopril, and enalapril in API, pharmaceutical dosage formulations, and human serum. Med
Chem Res. 2012; 21:4542. doi:10.1007/s00044- 012-9997-x.
42) Raj HA, Rajput SJ, Dave JB, Patel CN. Development and Validation of Two Chromatographic Stability-
Indicating Methods for Determination of Rosuvastatin In Pure Form and Pharmaceutical Preparation. Int
J ChemTech Res. 2009; 1(3):677-89. 43) Pandya CB, Channabasavaraj KP, Chudasama JD, Mani TT. Development and Validation of Rp-Hplc
Method for Determination of Rosuvastatin Calcium in Bulk and Pharmaceutical Dosage Form. 2010;
5(1)82-86.
44) Kaila HO, et al. A new improved RP-HPLC method for assay of rosuvastatin calcium in tablets. Indian
journal of pharmaceutical scienc. 2010; 72(5): 592.
45) Donthula S, Kumar MK, Teja GS, Kumar YM, Krishna JY, Ramesh D. A new validated RP-HPLC method
for determination of Rosuvastatin calcium in bulk andpharmaceutical dosage form. Der Pharmacia
Lettre. 2011; 3(3): 350-356.
46) Safwan A , Omar A. Validated high-performance liquid chromatographic method for the estimation of rosuvastatin calcium in bulk and pharmaceutical formulations. International journal of biomedical
science. 2011; 7(4): 283.
47) Trivedi HK, Patel MC. Development and Validation of a Stability-Indicating RP-UPLC Method for
Determination of Rosuvastatin and Related Substances in Pharmaceutical Dosage Form. Scientia
Pharmaceutica. 2012;80(2):393-406. doi:10.3797/scipharm.1201-09.
48) Mostafa Nadia M, et al. Selective Chromatographic Methods For The Determination Of Rosuvastatin
Calcium In The Presence Of Its Acid Degradation Products. Journal of Liquid Chromatography & Related
Technologies. 2014; 37(15): 2182-2196.
49) Smitha G, Sharath S, Reddy C, Sameer Kumar D, Shiva Kumar J, Jukanti Raju. Rosuvastatin calcium
Quantification in Rat Serum with the aid of RP-HPLC: Method Development and Validation. IOSR Journal
of Pharmacy and Biological Sciences. 2015;10(5): 23-28., DOI: 10.9790/3008-10522328
50) Kumar HT, Sri SD, Rao VPK and Rao SY. Validated RP-HPLC Method for Determination of Rosuvastatin
Calcium in Bulk and Pharmaceutical Formulation. Int J Pharm Sci Res. 2015; 6(7): 2913-17.doi:
10.13040/IJPSR.0975-8232. 6(7).2913-17.
Available Online : www.ijapbr.com
M. Hassouna et al IJAPBR, 2017; 2(2):11-27
27
51) Altınöz S, Uyar B. Electrochemical behaviour and voltammetric determination of rosuvastatin calcium in
pharmaceutical preparations using a square-wave voltammetric method. Ana Methods. 2013; 5: 5709-
5716.
52) Ramadan A, Mandil H, Ghazal N. Electrochemical Behavior and Differential Pulse Polarographic
Determination of Rosuvastatin In Pure Form and In Pharmaceutical Preparations Using Dropping Mercury Electrode . Int J Pharm Pharm Sci. 2014; 6(3): 128-133.
53) Baldut M, Bonafede SL, Petrone L, Simionato LD, Segall AI. Development and Validation of a
ComplexometricTitration Method for the Determination of Rosuvastatin Calcium in Raw Material. AIR.
2015; 5(5): 1-8.
54) ICH, Q2 (R1) Validation of Analytical Procedures: Text and Methodology. ICH Harmonized Tripartite
Guideline (2005).
Top Related