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Abstract

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF CLOMIPHENE CITRATE AND ACETYLCYSTEINE IN THEIR COMBINED TABLET DOSAGE FORM

Swati S. Agawane*, Ashpak M. Tamboli,Nazia I. Khan , Manoj S.Patil, Sneha S.Ghule, Swati T.Mane

Sahyadri College of Pharmacy, Methwade, Sangola, Solapur,Maharashtra, India-413307

ABSTRACT

The simple, accurate, rapid, and selective analytical procedures were developed and validated for the simultaneous estimation of Clomiphene citrate and Acetylcysteine using reverse phase High-performance liquid chromatography. The HPLC method employs a C18 Column (250mm x 4.6mm, 5m particle size 10 µL injection volume, column temperature controlled at 30° C, detection at 225nm as a isobestic point, Acetonitrile: Water (70: 30 v/v) is used in the mobile phase with a flow rate of 1ml/min. Parameters consisting of linearity, precision, accuracy, robustness, detection and quantitation are studied. From the investigations, the Absorptive factor of Clomiphene Citrate and Acetylcysteine at both the isosbestic points has been found at 225nm. Acetylcysteine had a 2.43minute retention duration, while Clomiphene Citrate had a 4.26 minute retention period. The detection concentrations for Acetylcysteine and Clomiphene Citrate were linear over the ranges of 240-1200 µg/mL and 20-100 µg/mL, respectively. Acetylcysteine and Clomiphene Citrate have regression equations of y = 147.9x +15846 and y = 187.1x + 6641, respectively with regression coefficients of 0.997 and 0.999. The percentage assay of 99.8% and 101.1% for Clomiphene Citrate and `Acetylcysteine. The recovery of Clomiphene Citrate and Acetylcysteine was found to be 99.6% and 101.3%. The objective of validation of analytical procedures is to demonstrate that it is suitable for its intended purpose.

Keywords: Clomiphenecitrate ; Acetylcysteine ; Simultaneous equation; Validation ; RP-HPLC.


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