Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHOD FOR DARUNAVIR IN PHARMACEUTICAL DOSAGE FORM

Prathamesh Dhiwar*, Rahul Dumbre and Narendra Gowekar

Abstract

High-performance liquid chromatography is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds. HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase (s) through the column, and a detector that shows the retention times of the molecules. Retention time varies depending on the interactions between the stationary phase, the molecules being analyzed, and the solvent (s) used. The sample to be analyzed is introduced in small volume to the stream of mobile phase and is retarded by specific chemical or physical interactions with the stationary phase. The amount of retardation depends on the nature of the analyte and composition of both stationary and mobile phase. The time at which a specific analyte elutes (comes out of the end of the column) is called the retention time. For the characterization of the drug some identification tests were done such as identification of drug, melting point of the drug and solubility test etc. methods were perform during entire process are as follows Analysis of tablet formulation of DAR in which determination of Recovery study of marketed formulation of DAR and Validation of RP-HPLC method for analysis of DAR along with Specificity, linearity, accuracy, precision, reproducibility, LOD and LOQ To access the stability of drug it is subjected to various stress conditions like acid, alkali, neutral oxidative and photo degredation these test were performed. For the assessment of different test dilution were prepared in a different concentration to obtain the accurate results dilutions were in range 0f 80- 240μg/ml.Developed and estimated stability indicating RP-HPLC method for darunavir which complies with ICH guidlines. Seperation was accomplished on agilent C18 colums (150 mm × 4.6mm, 3μm) using acetonitrile and phosphate buffer (40:60) as mobile phase. Flow rate of 1.5ml/min and optimised wavelength was 266nm. The calibration graph showed linear relationship with R=0.999 in range 0f 80-240μg/ml. To access the stability of drug it is subjected to various stress conditions like photo degradation, acid degradation, alkali degradation and oxidative degradation. With the conclusion from above information that the proposed HPLC method can be used successfully for estimation of Darunavir.

Keywords: Darunavir, ICH, Reverse phase High performance liquid chromatography, Forced Degradation, Development, validation.