DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF GLIPIZIDE AND MOMORDICININ IN RAT PLASMA

Momordicinin (MMR) is a major triterpenoid compound from Momordica charantia Linn fruit having antidiabetic potential, whereas glipizide (GPZ) is a well-known molecule in treatment of diabetes mellitus. The primary objective of the study is to develop and validate a simultaneous RP-HPLC method for upcoming pharmacokinetic studies to analyze herb-drug interactions. The study uses the protein precipitation method for plasma sample preparation. Chromatographic separation of GPZ and MMR was well achieved using hypersil gold C18 column (250 × 4.6, 5µm) and mobile phase constituting acetonitrile: phosphate buffer 10 mM with pH 3.5 (40:60, v/v). Quantification was done at wavelength 223 nm. The method was validated as per USFDA guidelines. The developed method showed good linearity over the range of 7-17 µg/ml, and the lower limit of quantifications was found to be 1.89 µg/ml and 1.99 µg/ml for MMR and GPZ, respectively. The extraction and chromatographic separation of both the drugs was found to be robust and reproducible. The %RSD for validation parameters was within the specified limits as per USFDA guidelines. The developed and validated bioanalytical RP-HPLC method for quantitative measurement of MMR and GPZ simultaneously was found to be accurate, rapid, simple, and reproducible, which will be applicable for future pharmacokinetic studies.