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Journal of Chemical and Pharmaceutical Research, 2012, 4(5):2404-2408

Research Article

ISSN : 0975-7384
CODEN(USA) : JCPRC5

Development and Validation of HPLC Method for the Determination of

Simvastatin in Bulk and Pharmaceutical Formulation
Praveen Kumar S. N*. and Bhadre Gowda D.G.
Department of Studies in Chemistry, Yuvarajas College, University of Mysore, Mysore,
Karnataka, India
______________________________________________________________________________
ABSTRACT
A simple, sensitive and validated HPLC method has been developed to determine Simvastatin in bulk drug and
pharmaceutical formulation. The chromatographic method was achieved on a C18 column (150x4.6 mm, 2.7 m)
using a mixture of methanol and 0.1% ortho phosphoric acid in water (10:90) at a wavelength of 238 nm. The
retention time of Simvastatin was found to be 3.106 min. The method was validated for specificity, linearity,
precision, accuracy and ruggedness. The result of robustness study also indicate that the method is robust and is
unaffected by small variation in chromatographic condition. The method is specific against excipient interference.
The method was found to show good linearity in the concentration range of 5.0 60.0 g/ml of Simvastatin with
correlation co-efficient of 0.9999. Accuracy was between 97.45% and 98.32%. Validation of the method was
carried out as per USP and ICH guidelines.
Key words: Simvastatin, HPLC, Validation.

______________________________________________________________________________
INTRODUCTION
Simvastatin belongs to the group of cholesterol-lowering lactones knows as statins which are among the worlds
most widely prescribed drugs. In addition to its use to control elevated cholesterol level it is also prescribed as a
preventive in cardiovascular diseases. It generally inhibits the production of mevalonate, which is responsible for the
endogenous synthesis of cholesterol, through competitive binding of 3-hydroxy-3-methylglutarylcoenzyme
reductase inhibitors [1]. The literature survey of Simvastatin revealed few methods based on chromatography [2-14]
have been reported for the determination. The scope and objective of present work describes the development and
validation of a precise, accurate, reproducible and economical reverse phase high performance liquid
chromatographic method with isocratic mode for the determination of Simvastatin in bulk drug and its
pharmaceutical formulation indicating the scientific novelty of the work. The proposed method was optimized and
validated in accordance with USP [15] and International Conference on Harmonization (ICH) guidelines [16].
EXPERIMENTAL SECTION
Apparatus and Chromatographic Conditions: Agilent 1200 series HPLC, G1331A Quaternary pump connected
with G1314B Variable Wavelength detector, G1316A Thermostatted Column Compartment, G1329A ALS
autosampler. The data acquisition was performed by Agilent Chemstation software. Analysis was carried out with
Poroshell SB C18 (150x4.6 mm) 2.7 m column, using isocratic elution. The run time was 6.0 min with a flow rate

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J. Chem. Pharm. Res., 2012, 4(5):2404-2408
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of 1.0 ml/min. UV detection was performed at 238 nm. Peak identity was confirmed by retention time comparison
and the HPLC was operated at room temperature.
Reagents: ortho phosphoric acid (AR grade, Spectrochem), methanol (HPLC grade, Thomas Baker), water (HPLC
grade, Thomas Baker) and Simvastatin.
Preparation of Mobile Phase:
The mobile phase is composed of a mixture of methanol and 0.1% ortho phosphoric acid in water 10:90 (v/v),
filtered through a 0.45 m nylon filter (Millipore, USA) and degassed by sonication prior to use to avoid
disturbances and column clogging due to small particles..
Preparation of Standard Solution:
The standard stock solution of Simvastatin (0.5 mg/ml) was prepared in methanol. The working standard solution
Simvastatin (50 g/ml) was prepared by diluting the stock solution with mobile phase solution.
Preparation of Sample Solution:
Twenty tablets were weighed to get the average weight and then ground. An amount of powder equivalent to 50 mg
of Simvastatin was transferred to a 100 ml volumetric flask, added 70 ml of methanol and sonicated for 30 min with
intermediate shaking. Then making up to volume with methanol to obtain a solution containing 0.5 mg/ ml
Simvastatin. An aliquot was then removed and centrifuged for 10min. The solution was filtered using 0.45m
membrane filter paper. After filtration, the solutions were diluted with mobile phase to give a final concentration of
Simvastatin (50 g/ml) and 10.0 l of clear solution was injected into HPLC system for the analysis.
System Suitability:
The column was equilibrated with mobile phase at prescribed condition till a stable baseline was achieved.
Separately injected the single injection of diluent, six injections of standard and duplicate injections of sample and
recorded the chromatogram.
The following system suitability criteria were fulfilled in the chromatogram obtained with standard preparation:
% Relative standard deviation (RSD) for Simvastatin peak area of replicate injections should not be more than
2.00.
Tailing factor for Simvastatin peak should be between 0.80 - 2.00.
The theoretical plates for Simvastatin peak should not be less than 2000.
RESULTS AND DISCUSSION
Method Development:
The object of this work was to develop and validate a simple, rapid and sensitive assay method for the determination
of Simvastatin. To achieve the objective, different options were evaluated. Optimization of the chromatographic
conditions is intended to take into account that various goals of method development like runtime sensitivity, peak
symmetry, theoretical plates, tailing factor, etc. The chromatographic conditions were optimized by changing the
mobile phase compositions and buffers used. Finally a mixture of methanol and 0.1% ortho phosphoric acid in water
in the ratio of 10:90 was used. The retention time of Simvastatin was 3.106 min with C18 column (150 x 4.6 mm
i.d.2.7m) and UV detection 238nm. A typical chromatogram obtained by using the aforementioned mobile phase
from 10.0 l of the assay preparation is illustrated in Figure 1.
Accuracy:
The recovery studies were carried out for the accuracy parameter. The recovery was determined at three levels, viz.,
50%, 100% and 150% of the selected concentrations and percentage recovery for Simvastatin ranges from 97.4598.32% indicates that developed method is accurate and reliable the data are tabulated inTable-1.

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Figure 1: A typical chromatogram showing the peak of Simvastatin

Table -1: Recovery Study of Simvastatin.

Set No.
1
2
3

1
2
3

1
2
3

Accuracy
Levels (%)
50

100

150

Amount
Added (mg)
25.2
25.4
25.3

50.2
50.1
50.2

75.4
75.6
75.3

Amount
Recovered (mg)
24.88
24.86
24.88
Mean
RSD %
49.39
49.92
48.91
Mean
RSD %
73.48
73.59
73.46
Mean
RSD %

Amount
Recovered (%)
98.74
97.88
98.35
98.32
0.44
98.38
97.64
97.44
97.82
0.51
97.46
97.34
97.56
97.87
0.35

Precision:
The precision (repeatability and intermediate precision) of the method was determined from one lot of combined
dosage form. Inter-day and intra-day studies were performed by taking six replicates of assay preparation and %
RSD was found to be less than 2, tailing factor and the theoretical plates found to be well within the system
suitability limits are tabulated in Table-2.
Linearity:
It was observed that the optimized method were linear within a specific concentration range of Simvastatin. The
calibration curves were plotted between response factor and concentration of the standard solutions. The linearity
ranges were found to be 5.0- 60.0 g/ml with correlation coefficients of 0.9999 for Simvastatin as shown in Figure
2.

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Table -2: System Suitability Test of Simvastatin.
Sl. No.
1
2
3
4
5
6
Average
Standard Deviation
RSD %
Tailing Factor
Theoretical Plates

Simvastatin
Simvastatin
Retention Time
Peak Area
3.107
1093.4
3.103
1094.2
3.103
1091.9
3.105
1093.4
3.105
1094.0
3.110
1093.5
3.106
1093.4
0.003
0.807
0.09
0.07
1.138
13845

Figure 2: Graph showing Linearity and Range of Simvastatin.

Solution Stability:
Stability studies were carried out by keeping the prepared sample solution at room temperature for 24 hours. The
prepared sample solution was analyzed at different time intervals and found that the solution is stable.
Robustness:
The assay results were found to be unaffected by small changes in the chromatographic conditions like
concentration of the mobile phase, flow rate and wavelength.
Ruggedness:
Ruggedness of the method was determined between two different analysts and instruments. The value of % RSD
was < 2, tailing factor and the theoretical plates found to be well within the system suitability limits. This indicates
the ruggedness of the developed analytical method.
LOD and LOQ:
For calculating the LOD and LOQ values, solutions with known decreased concentrations of analytes were injected
into the HPLC system. The limit of detection (LOD) and quantification (LOQ) were then measured by calculating
the minimum level at which the analytes can be readily detected (signal to noise ratio of 3:1) and quantified (signal
to noise ratio of 10:1) , the LOD was found to be 0.25 g/ml and LOQ was found to be 5.0 g/ml Simvastatin
respectively.

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J. Chem. Pharm. Res., 2012, 4(5):2404-2408
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Application of the developed method:
The method is sensitive and specific for the determination of Simvastatin and also subjected to validation for
different parameters, hence has been applied for the estimation of drug in bulk and its pharmaceutical formulation.
The amount of Simvastatin was found to be within the range of 95% - 105%.
CONCLUSION
The proposed method was found to be simple, precise, accurate and rapid for the determination of Simvastatin in
bulk drug and its pharmaceutical formulation by isocratic mode. The mobile phase is simple to prepare and
economical. The validation of the analysis proved that the method is reproducible and efficient for the determination
of Simvastatin without any interference from the excipients. The HPLC method was validated and demonstrated
good linearity, precision, accuracy and specificity. Thus, the developed method can be easily and conveniently
utilized for routine analysis in quality control laboratories for the determination of Simvastatin.
Acknowledgements
The authors are thankful to Dr. Ravi Datar, R&D Manager, FMC India R&D Center, Indian Institute of Science
Campus, Bangalore, Karnataka, India, for providing facilities to carry out this work.
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