Development and validation of RP HPLC method for the estimation of Sofosbuvir and related impurity in bulk and pharmaceutical dosage form

Abstract

Background The present work is aimed at development and validation of RP HPLC method which is simple, specific, precise, and accurate for estimation of Sofosbuvir and its process-related impurity in bulk and pharmaceutical dosage forms. Extensive literature survey revealed no method for estimation of the above said. The chromatographic separation was achieved on Agilent Eclipse XDB-C18, 4.6 × 250 mm, 5 μm with mobile phase composed of 0.1% trifluoroacetic acid in 1000 ml of water:acetonitrile (50:50) using an isocratic mode of elution. Detection was made using UV detector at 260.0 nm and LC solution software for analysis of data. The developed method was validated according to ICH guidelines. Results The linearity of calibration curve for Sofosbuvir in concentration range of 160-480 μg/ml was good. The curve was linear for its process related impurity (Phosphoryl) in concentration range of 10-30 μg/ml. There exists a good correlation between peak area and analyte concentration. Retention time for Sofosbuvir was found to be 3.674 min and its impurity was 5.704 min. Relative standard deviation values for Sofosbuvir is 1.741 and its process related impurity is 0.043. LOD for Sofosbuvir and its impurity was found to be 0.01% (0.04 μg) and 0.03% (0.12 μg) respectively. LOQ for Sofosbuvir and its impurity was found to be 0.50% (0.125 μg) and 1.50% (0.375 μg) respectively. Conclusion All the results reveal that the proposed method was found to be highly sensitive, simple, precise, accurate, robust, and fast. Large number of samples can be analyzed in shorter time due to shorter retention times, so it can be successfully applied for routine analysis of Sofosbuvir and related phosphoryl impurity in bulk and pharmaceutical dosage forms.

RP-HPLC Method Development and Validation for an Estimation of Empaglifl ozin from Bulk and Pharmaceutical Dosage Form

“…Estimates of the correlation coefficient and intercept were derived from the calibration curve's corresponding equation of the line. [32][33][34] The equation that describes a straight line in its most generic form is shown below. Y = mX+c Where, Y = Peak region; m = slope; X = measured concentration; c = intercept.…”

“…In each instance, the percent RSD was computed, and then conformity with the stipulated requirements was determined by examining the results. [32][33][34] %Accuracy According to the precision study, the accuracy of the procedure was decided by utilizing three different standard solutions of the empagliflozin that were included in the combination (API mixture). After injecting the solutions three times, the concentration of each solution was calculated by extrapolating from the calibration curve.…”

“…After that, an estimate of the accuracy percentage was calculated using the subsequent formula. [32][33][34] Limit of Detection and Limit of Quantitation A sample's LoD is the lowermost amount of analyte that can be recognized through a given assay, whereas a sample's LoQ is the lowermost amount of analyte that can be determined. The following formula was used to obtain these values for the parameters.…”

“…The following formula was used to obtain these values for the parameters. [32][33][34] Where, s = Standard deviation and S = Slope of the calibration curve.…”

Analytical Method Development and Validation of Rp-HPLC Method for the Estimation of Sofosbuvir in Bulk and Formulation

“…The pharmacokinetics of Sofosbuvir and predominant circulating metabolite GS-331007 have been evaluated in healthy subject and subject with chronic hepatitis C. Oral administration doses up to 400mg given to both subjects, Sofosbuvir was absorbed with peak plasma concentration observed at ~0.5-2 hours post dose and regardless of the dose level and GS-331007 peak was observed between 2-4 hours post dose [7]. From literature survey, it was found that few methods are available for determination of single drug and in combination containing Sofosbuvir and Ledipasvir by UV-VIS spectroscopes [8][9][10][11], RP-HPLC methods [11][12][13][14][15][16][17][18]. The aim of the study is to develop and validate a sample, precise and accurate RP-HPLC method for Sofosbuvir in bulk and formulation as per ICH guidelines.…”