Industrial Chemistry
Open Access

Rosuvastatin calcium is used as an antihyperlipidemic, which is a HMG-CoA reductase inhibitor, a rate limiting enzyme in cholesterol biosynthesis. It is used in the treatment of dyslipidemia, which is effective at low doses and its half-life is more compared to other statins. Chemically it is, calcium salt of (3R,5S,6e)-7-(4-(4-Fluorophenyl)-6-(1-methylethyl)-2-(ethyl(methylsulfonyl)amino)-5-pyrimidinyl)-3,5-dihydroxy-6-heptenoic acid and molecular formula is (C₂₂H₂₇FN₃O₆S)₂Ca (Figure 1) [1]. Extensive literature survey reveals that a few spectrometric were available for the estimation of Rosuvastatin calcium in bulk and formulations [2-9]. The objective of the present study is to develop a new simple, sensitive, accurate, rapid and economic method for the estimation of Rosuvastatin calcium in bulk and tablet formulation.

Figure 1: Structure of Rosuvastatin.

Literature survey reveals that only an LC-MS study has been reported for the quantification of Rosuvastatin calcium in human plasma. Some UV spectrophotometric method has been reported for the estimation of Rosuvastatin calcium. The objective of the present work was to develop a simple, sensitive, precise and accurate UV spectrophotometric method for the determination of Rosuvastatin calcium in bulk and pharmaceutical formulations as per ICH Guidelines [10].

Materials and Methods

Instruments

A Shimadzu UV-visible spectrophotometer (UV 1800, Shimadzu Corporation, Kyoto, Japan) was used for all absorbance measurements with matched quartz cells.

Chemicals and reagents

All chemicals acetonitrile ammonium acetate and reagents were of analytical grade. Rosuvastatin calcium in the form of powder with certificate of analysis was provided by Cipla Research Centre, Mumbai. Pharmaceuticalgrade excipients were obtained from Pharmaceutical Technology Lab of Maharashtra.

Results

Determination of maximum wavelength (λmax)

Standard stock solution of 100 μg/ml of Rosuvastatin calcium was prepared in mobile phase. From the above stock solution, pipette out 1 ml into 10 ml volumetric flask and finally made up the volume with mobile phase to produce a concentration of 10 μg/ml. The samples was then scanned in UV spectrophotometer from a range of 200-400 nm against mobile phase as blank and the wavelength corresponding to maximum absorbance in methanol was found at 242 nm (Figure 2).

Figure 2: UV spectrum of Rosuvastatin calcium.

Preparation of standard calibration curve

For the preparation of standard calibration curve, use stock of 100 μg/ml concentrations and 2-12 μg/ml concentration were prepared for analyzing calibration curve by pipetting out 0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 ml from the stock solution into a 10 ml volumetric flask and made up the volume with mobile phase [11-13]. The absorbance of each solution was measured at 242 nm as blank. Calibration curve of the drug was then plotted by taking the absorbance obtained on y-axis and the concentration of the solution on x-axis (Figure 3).

Figure 3: Calibration curve of Rosuvastatin.

Validation

Validation can be defined as (ICH) establishing documented evidence, which provides a high degree of assurance that a specific activity will consistently produce a desired result or product meeting its predetermined specifications and quality characteristics.

The method was validated for several parameters like linearity, accuracy, precision, ruggedness, robustness, Limit of Detection (LOD), and Limit of Quantification (LOQ) according to ICH guidelines.

Linearity

The linearity of the analytical method was its ability to elicit test results which are directly proportional to analyte concentration in samples within a given range. To establish the linearity of the proposed method, various aliquots ofthe standard solution of the drug were prepared from stock solution and analyzed [14]. The drug showed linearity in therange of 2-12 μg/with correlation coefficient 1. Linearity data are shown (Table 1 and Table 2).

Table 1: Linearity table of Rosuvastatin calcium.

Table 2: Linear regression data for calibration curve.

Accuracy

This study was carried out using the stock solution (100 μg/ml). Take three concentrations 2 μg/ml, 6 μg/ml, and 12 μg/ml. And take six reading of these concentrations [15]. The result was reported as % RSD. The accuracy result showed a good with percent relative standard deviation less than 2. A result shows that the method is accurate up to 12 μg/ml.

Precision

Precision studies were carried out to ascertain the reproducibility of the proposed method. Repeatability was determined by preparing six replicates of same concentration of the sample and the absorbance was measured.

Intraday precision study was carried out by preparing drug solution of same concentration and analyzing it at three different times in a day. The same procedure was followed for one day to determine interday precision.

The result was reported as % RSD. The precision result showed a good reproducibility with percent relative standard deviation less than 2. The result shows that the method was found to be precise.

Robustness

Analysis was carried out at two different wavelengths to determine the robustness of the method and the respective absorbance was measured. The result was indicated as % RSD which is less than 2%. The results show that the method is robust for change in wavelength.

LOQ and LOD

Limit of Detection (LOD) is the lowest amount of analyte in the sample that can be detected. Limit of Quantification (LOQ) is the lowest amount of analyte in the sample that can be quantitatively determined by suitable precision and accuracy. The values of LOQ and LOD were found to be 0.769306 and 2.331231 μg/ml respectively.

Degradation Studies

The ICH guidelines Q1A (R2) entitled stability testing of new drug substances and products that required stress testing to be carried out to elucidate the inherent stability characteristics of the active substance. The aim of this study was to perform the stress degradation studies on Rosuvastatin calcium using the developed method.

Stress degradation by hydrolysis under acidic condition

The preparation of 0.01 N Hydrochloric Acid (HCL) was done by diluting 0.085 ml of conc. HCL to 100 ml of distilled water. Amount of drug was weighed and was transferred to a labeled round bottomed flask. Refluxed the sample for 2 hrs and pipette out 1 ml to 10 ml volumetric flask, made up the volume with mobile phase to get the final dilution so as to get final test concentration. Taken the reading on UV. Check the degradation behavior of drug.

Stress degradation by hydrolysis under alkaline condition

The 0.01 N Sodium Hydroxide (NaOH) was prepared by dissolving 0.04 gm of sodium hydroxide pellets in 100 ml of distilled water. Amount of drug was weighted and was transferred to a labeled round bottom flask. Reflux the sample for 2 hrs. and pipette out 1 ml to 10 ml volumetric flask, made up the volume with diluent to get the final dilution so as to get final test concentration to get the final test concentration 10 ppm. Take the reading on UV. Check the degradation behavior of drug.

Stress degradation by neutral condition

Amount of drug was weighed and transferred in to 100 ml water in round bottom flask. Reflux it for 2 hours. Pipette out 1 ml in to 10 ml volumetric flask, made up the volume with diluent to get the final dilution so as to get final test concentration 10 ppm. Take the reading on UV. Check the degradation behavior of drug.

Photolytic degradation

Photo stability was performed by placing amount of drug in daylight for 24 hours. The samples were diluted with mobile phase up to 10 ml in a volumetric flask. Pipette out 1 ml sample diluted up to 10 ml by diluent to get the final dilution so as to get final test concentration 10 ppm. Take the reading on UV. Check the degradation behavior of drug.

Dry heat induced degradation

Standard drug was placed in an oven at 60°C for 2 hours to study dry heat degradation. Amount of drug samples were diluted with mobile phase up to 100 ml in a volumetric flask. Pipette out 1 ml and were diluted up to 10 ml by diluent to get the final dilution so as to get final test concentration 10 ppm. Take the reading on UV. Check the degradation behavior of drug.

Oxidative degradation

Weight accurately amount of drug and transfer into 10 ml of volumetric flask which containing 3% hydrogen peroxide. Keep it in dark place for 2 hours. Pipette out 1 ml sample diluted up to 10 ml by diluent to get the final dilution so as to get final test concentration 10 ppm. Take the reading on UV. Check the degradation behavior of drug(Table 3).

Table 3: % Degradation under stress conditions.

Conclusion

The proposed UV method is simple, accurate, precise, specific and highly sensitive. The method is economical rapid and do not require any sophisticated apparatus in contrast to spectroscopic methods. The proposed method is also useful for the determination of Rosuvastatin calcium stability in samples of pharmaceutical dosage forms. Hence, the proposed method can be successfully used for routine quality control analysis of drug in marketed preparations.

Acknowledgment

The authors take this opportunity to thanks Cipla Research Centre, Mumbai for providing gift sample. The authors would like to acknowledge Principal Dr. Sanjay Thonte, Channabasweshwar Pharmacy College, Latur for providing facilities for conducting research.

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