INTRODUCTION:

Rosuvastatin Calcium² (3R,5S,6E)-7-[4-(4-Flurophenyl)-2-(N-methylmethanesulphanamido)-6-(propan-2-yl) pyrimidin-5-yl]-3,5-dihydrohept-6-enoic acid is an antihyper lipidemic drug belong to the class of medications called HMG-CoA reductase inhibitors. It works by retarding the cholesterol production in the body and is used to prevent Cardiovascular diseases.

Rosuvastatin Calcium² (3R, 5S, 6E)-7-[4-(4-Flurophenyl)-2-(N-methylmethanesulphanamido)-6-(propan-2-yl) pyrimidin-5-yl]-3, 5-dihydrohept-6-enoic acid is an antihyper lipidemic drug belong to the class of medications called HMG-CoA reductase inhibitors.

It works by retarding the cholesterol production in the body and is used to prevent Cardiovascular diseases.

Figure 1. Structure of Rosuvastatin Calcium ²

Fenofibrate Propan-2-yl[4-(4-chlorobenzol)phenoxy]-2-methyl propanoic acid,³ 1methylethyl ester is an anti-hyper lipidemic drug which activates peroxisome proliferator activated receptor alpha. This in turn activates lipoprotein lipase and reduces the production of apolipoprotein.

Figure 2. Structure of Fenofibrate¹

The objective of this study was to develop and validate a simple and specific Zero cross technique for the simultaneous determination of Rosuvastatin Calcium and Fenofibrate⁵ in combined dosage form. This is one of the derivative spectroscopic techniques that is widely used as a tool for quantitative analysis. This technique offers various advantages over the conventional spectrophotometricmethods^6,7, such as discrimination of the sharp spectral features over the large bands and enhancement of the resolution of the overlapping spectra^8,9. This proposed method was aimed at exhibiting a precise, accurate and cost effective simultaneous assay for the selected drugs.

MATERIALS AND METHODS:

Instruments and materials:

The proposed UV Spectroscopic method was carried out using Agilent Cary 60 UV-Visible double beam Spectrophotometer with 1cm matched quartz cells. Rosuvastatin Calcium and Fenofibrate were procured from Yarrow Chem Products, Mumbai. A Commercially available brand (ROSAVE F 10) containing 10mg of Rosuvastatin Calcium and 160mg of Fenofibrate was purchased from a local market in Vizianagaram, Andhra Pradesh. Ethanol^10,11 was purchased from Fisher Scientific, Mumbai.

Preparation of standard stock solution:

10 mg of standard Rosuvastatin Calcium^12,13 and Fenofibrate were weighed separately, transferred to 100 ml volumetric flasks and dissolved in ethanol. The flasks were shaken and volumes were made up to the mark with ethanol to give a solution containing 100µg/ml each of Rosuvastatin Calcium and Fenofibrate.

Methodology:

The working standard solutions of Rosuvastatin Calcium¹⁴ and Fenofibrate were prepared separately in ethanol both having concentration range of 5-25µg/ml. Suitable aliquots of the selected drugs were scanned in the wavelength range of 200-400 nm against solvent ethanol as blank and the absorption spectra was obtained as shown in figure 3. This Zero order spectrum was derivatised to 1°, 2° and 3° spectra. Third order (3°) derivative spectrum was selected for the analysis of both drugs. From the overlain spectra of both the drugs, wavelengths selected for quantitation were 235, 251 and 268nm (Zero cross points for Rosuvastatin Calcium) and 249, 273 and 291nm (Zero cross points for Fenofibrate).

Figure 3. Isosbestic point of Rosuvastatin Calcium and Fenofibrate

Figure 4. Third order derivative spectrum of Rosuvastatin Calcium and Fenofibrate

RESULTS AND DISCUSSION:

The developed method was validated using different validation parameters like accuracy, precision, LOD, LOQ, robustness and ruggedness as per International Council for Harmonization guidelines.

Validation Parameters¹⁵:

Linearity:

Appropriate aliquots from the standard stock solution of Rosuvastatin Calcium and Fenofibrate were used to prepare two different sets of dilutions. Series A consisted of working standard solutions of Rosuvastatin Calcium in the concentration range of 5-25µg/ml. Series B consisted of varying concentrations of Fenofibrate in the range of 5-25µg/ml. These working standards were analysed by the proposed method. The calibration curves for Rosuvastatin Calcium and Fenofibrate obtained were assessed from their respective regression equations. The responses were found to be linear over the abovementioned range with correlation co-efficients of 0.9993 and 0.9992 for Rosuvastatin Calcium and Fenofibrate, respectively.

Accuracy:

The accuracy of the method was determined by calculating the recoveries of Rosuvastatin Calcium and Fenofibrate by the standard addition method. Known amounts of standard Rosuvastatin Calcium and Fenofibrate were added at 80%, 100% and 120% levels to the pre-analysed sample solutions of Rosuvastatin Calcium and Fenofibrate.

Table 1. Accuracy Data

Drug Name

% Added

Amount of drug added (mg)

Drug Recovered (mg)

% Recovery

Mean

Rosuvastatin Calcium(5mg)

80%

100%

120%

8.58

10.23

11.22

95.33

102

99.7%

Fenofibrate(32mg)

80%

100%

120%

57.52

65.4

69.63

99.1

102

99.4

100%

Table 2. Precision Data

Intra day

Inter day

Drug Name

Absorbance

% RSD n=3

Mean

Absorbance

% RSD n=3

Mean

Rosuvastatin Calcium

5µg/ml

0.348

0.358

0.341

0.0632

0.0831

0.0910

0.0936%

0.431

0.423

0.434

0.083

0.102

0.132

0.105%

Fenofibrate

32µg/ml

1.4024

1.3991

1.4029

0.1471

0.5796

0.8016

0.679%

1.451

1.489

1.442

0.412

0.762

0.997

0.732%

The amounts of Rosuvastatin Calcium and Fenofibrate recovered were estimated by substituting the obtained values in the respective regression equations. The results are shown in Table 1.

Precision:

The reproducibility of the proposed method was determined by performing the assay on the same day (intraday precision) and on three consecutive days (interday precision). Precision was assessed from nine determinations (3 x 3 replicates) for each concentration covering the specified range. The low %RSD values show that the method is quite precise, as shown in Table 2.

LOD and LOQ:

The standard solutions of Rosuvastatin Calcium (5, 10, 15, 20 and 25μg/ml) and Fenofibrate (5, 10, 15, 20, 25 μg/ml) were analysed using the developed method. The minimum detectable and quantifiable limits were found. The standard deviation values of y-intercept in the regression equations were used to calculate LOD and LOQ from the expressions, LOD = 3.3 x SD/S and LOQ = 10 x SD/S. The low values obtained as shown in Table 3indicate that the proposed method is quite sensitive.

Table 3. LOD and LOQ

Parameters	Rosuvastatin Calcium(µg/ml)	Fenofibrate (µg/ml)
LOD	0.1524	1.0626
LOQ	0.4618	3.22

Robustness, Ruggedness:

The robustness of the developed derivative spectroscopic method was assessed by a slight variation in the wave length (±1). The ruggedness of the proposed method was determined by analysing the standard and sample solutions by two different analysts and also at two different laboratories to check the reproducibility. The results were summarised as shown in Table 4.

Table 4. Robustness, Ruggedness

	Robustness		Ruggedness
Drug Name	Wavelength 229±1 nm	Wavelength 270 ±1nm	Analyst	Laboratory
Rosuvastatin % RSD	0.949%	0.0954%	0.95%	0.098%
Fenofibrate % RSD	0.672%	0.703%	0.663%	0.724%

Assay of ROSAVE F 10 tablets.

Accurately weighed quantity equivalent to 1mg Rosuvastatin Calcium and 16mg fenofibrate of tablet powder was transferred into 100ml volumetric flask, 20ml of ethanol was added to it and sonicated for 30mins. The resulting solution was made up to the mark with ethanol and filtered. The filtrate was diluted suitably with ethanol to get a solution containing 10µg/ml of Rosuvastatin Calcium and 160µg/ml of Fenofibrate. Then 1ml of resulting solution was transferred to 10ml volumetric flask and diluted up to the mark with ethanol. The absorbance of the resulting solution was measured at 249nm (isosbestic point). The concentrations of Rosuvastatin Calcium and Fenofibrate present in the tablet solution were calculated using the regression equation of the respective drugs. The results are shown in Table 5.

Table 5. Assay Result of Marketed Formulation

Brand Name	Rosuvastatin Calcium(10mg)	Fenofibrate (160mg)
ROSAVE F 10	101.6%	100%

CONCLUSION:

A new Zero Cross technique for simultaneous estimation of Rosuvastatin Calcium and Fenofibrate in tablets by a UV derivative spectroscopic method was developed. The developed method was performed as per ICH guidelines. The proposed UV spectrometric method for the quantitative determination of Fenofibrate and Rosuvastatin Calcium in combined dosage form is found to be simple, rapid, precise, and accurate. The non-interference of the excipients in the analysis of the formulation indicates the specificity of the method. Therefore, the developed method can successfully be used for routine analysis of selected drugs in their combined dosage forms.

ACKNOWLEDGEMENT:

We are very much thankful to Maharajah’s College of Pharmacy, Vizianagaram, for giving us permission to carry out my research work.

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Received on 12.09.2022 Modified on 17.10.2022

Asian J. Pharm. Ana. 2023; 13(2):99-102.

DOI: 10.52711/2231-5675.2023.00017