Review On the Development, Validation, And Analytical Methods for Rosuvastatin: A Comprehensive Approach to Cardiovascular Treatment

Rosuvastatin is chemically designated as (3R,5S,6E)-7-[4-(4-fluorophenyl)-2-(N-methylmethanesulfonamido)-6-(propan-2-yl)pyrimidin-5-yl]-3,5-dihydroxyhept-6-enoic acid. It belongs to the statin class of drugs and is widely used to treat hypercholesterolemia and related conditions while preventing cardiovascular diseases. The molecular formula of Rosuvastatin calcium is (C22H27FN3O6S)2Ca(C_{22}H_{27}FN_3O_6S)_2Ca(C22?H27?FN3?O6?S)2?Ca, with a molecular weight of 1001.1 g/mol.

Several studies have focused on its estimation and stability in various forms. For example, Sonu Sundd Singh and Kuldeep Sharma (2005) employed an HPLC tandem mass spectroscopic method for Rosuvastatin estimation in human plasma, which has been applied in bioequivalence studies. Similarly, Marothu Vamsi Krishna and Dannana Gowri Sankar (2007) developed extractive spectrophotometric methods using Safranin O and Methylene Blue for determining Rosuvastatin calcium in both pure form and pharmaceutical formulations. Stability-indicating methods, such as those described by Nadia M. Mostafa, Amr M. Badawey, et al. (2012), have been utilized to identify oxidative degradation products and ensure the compound's stability during analysis.

Analytical methodologies for Rosuvastatin involve defining techniques, procedures, and protocols to meet specific analytical requirements, including sensitivity, accuracy, range, and precision. Rosuvastatin functions as a competitive inhibitor of HMG-CoA reductase, acting primarily in the liver. Its plasma elimination half-life is approximately 19 hours, contributing to its efficacy.

Validated analytical methods, developed in compliance with International Council for Harmonisation (ICH) guidelines, have demonstrated high accuracy, sensitivity, specificity, and reproducibility for Rosuvastatin analysis. These methods ensure consistent quality control and reliable outcomes for pharmaceutical applications.

2.0 MATERIALS AND METHODS

 2.1 Instrument Specifications

The study utilized a range of high-precision instruments to ensure accurate and reliable results. A Shimadzu AUX-200 digital balance was employed for precise weighing, while spectrophotometric analyses were conducted using both a Shimadzu 1700 double-beam UV-visible spectrophotometer and an Elico SL-210 double-beam UV-visible spectrophotometer, each equipped with a pair of 10 mm matched quartz cells. High-performance liquid chromatography (HPLC) analyses were carried out using a Shimadzu HPLC system (LC-10ATVP). Sample preparation involved the use of a Remi centrifuge apparatus and a Sonicator Model 2120 MH for effective ultrasonic treatment and thorough mixing. A Cyberlab micropipette was used for handling small liquid volumes with precision, and an Elico LI 120 pH meter ensured accurate pH measurements. Additionally, a melting point apparatus from Guna Enterprises, Chennai, was utilized to determine the melting points of substances, further supporting the study's analytical rigor.

2.2 Reagents and chemicals used in the study

All the chemicals used in the study were of analytical reagent grade and HPLC grade, procured from Qualigens India Pvt. Ltd., Mumbai. The chemicals included distilled water, acetonitrile (HPLC grade), methanol (spectral and HPLC grade), water (spectral and HPLC grade), and orthophosphoric acid (analytical grade).

2.3 Preparation of standard stock solutions : 

To prepare the desired concentration of the active pharmaceutical substance (ASP), an accurately weighed quantity of 75 mg of ASP was transferred into a 100 mL volumetric flask. Methanol was added to dissolve the substance completely, and the solution was diluted up to the mark with methanol to ensure a consistent and homogeneous stock solution.

From this stock solution, a 5 mL aliquot was carefully measured using a pipette and transferred into a separate 50 mL standard volumetric flask. Methanol was again added to dilute the solution to the mark, resulting in a final concentration of 75 µg/mL. This stepwise dilution process ensures precision and accuracy in achieving the desired concentration for subsequent analytical or experimental procedures.

2.4 Selection of wavelengths for estimation and stability studies :

Solutions of ASP and ROSU, each with a concentration of 10 ?g/mL, were scanned individually in the UV region of 200–400 nm using a spectrophotometer. The overlaid spectrum of ASP and ROSU in methanol was also recorded. From the overlain spectrum, the wavelengths 294.5 nm (the ?max of ASP) and 243 nm (the ?max of ROSU) were identified and selected for the Simultaneous Equation Method. For the Absorbance Ratio Method, the selected wavelengths were 243 nm (the ?max of ROSU) and 229.8 nm (the iso-absorptive point of ASP and ROSU). This systematic selection of wavelengths ensures accurate and reliable quantification of both compounds in simultaneous analysis.

2.5 Preparation of calibration graph :

From the above stock solution, aliquots were drawn and appropriately diluted to achieve final concentrations in the range of 7.5–37.5 ?g/mL for ASP and 1–5 ?g/mL for ROSU. The absorbances of these 30 solutions were recorded at their respective wavelengths using the specified methods. This step ensured accurate preparation and measurement of the calibration standards for subsequent analysis.

2.6 Recovery studies :

A recovery experiment was conducted to validate the accuracy of the analytical method by adding known concentrations of ASP and ROSU raw materials to a 50% pre-analyzed formulation. Standard solutions of ASP and ROSU, corresponding to 80%, 100%, and 120% of their respective theoretical concentrations, were added to the pre-analyzed formulation in a series of 10 mL volumetric flasks. Methanol was then added to dissolve the contents, and the solutions were diluted to the mark with methanol to ensure uniformity. This procedure allowed for the assessment of the method's ability to accurately recover the added analytes from the formulation.

2.7 Method Validation

(2.71) Linearity: Linearity was evaluated by diluting the standard stock solution to prepare five different concentrations. ASP demonstrated linearity within the concentration range of 7.5–37.5 µg/mL, while ROSU showed linearity between 1–5 µg/mL. Calibration curves, representing the mean values of six determinations, were plotted by correlating the concentration with the corresponding absorbance for both drugs.

(2.72) Precision: The precision of the method was assessed through repeatability and intermediate precision. Repeatability was tested by analyzing the formulation six times at the same concentration. The amount of each drug in the capsule formulation was calculated, and the % relative standard deviation (%RSD) was determined. Intermediate precision was verified through intraday and interday analyses. The amount of each drug and the %RSD for both analyses were calculated to ensure consistency and reliability of the method.

(2.73) Ruggedness: Ruggedness refers to the reproducibility of test results under expected operational conditions, including variability between laboratories and analysts. In this study, the ruggedness of the method was tested by determining ASP and ROSU concentrations under varying conditions, ensuring the results were consistent and reliable across different settings and operators.

2.8 RESULTS

The methods employed for the analysis of Rosuvastatin was in Table Dosage form

Figure-1 Calibration Curve Of   Rosuvastatin In Methanol At 294.5 Nm

       
           
       
Figure-2 Calibration Curve Of Rosuvastatin In Methanol At 229.8 Nm

Stability Studyof Rosuvastatin Calcium For Uv Spectroscopic Methods

Table-2 Intraday And Interday Analysis Of Formulation [Rozucor Asp-10]

(Simultaneous Equation Method)