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Abstract

DEVELOPMENT AND VALIDATION OF ANALYTICAL LC-MS METHOD FOR SIMULTANEOUS DETERMINATION OF ORAL HYPOGLYCEMIC AGENTS(MET, DFZ, EFZ AND CFZ) IN PHARMACEUTICAL DOSAGE FORM

Mohini A. Patel*, Dr. Pranav Pandya, Dr. Seema Kothari, Dr. Suresh Chanda Ameta

ABSTRACT

A simple, rapid and highly sensitive and selective liquid chromatography-mass spectrometry (LC–MS) method was developed and validated for the simultaneous estimation of the recently approved oral hypoglycemic mixture of metformin (MET), canagliflozin (CFZ), empagliflozin (EFZ) and dapagliflozin (DFZ) in pharmaceutical dosage form. The sepration was carried out on column Acquity BEH C18 (50mm, 2.5mm, 5μm) as mobile phase of Line A: 5mM ammonium acetate in 0.1%formic acid in water and Line B:0.1% formic acide in acetonitrile. The isocratic ratio A:B=10:90v/v with flow rate 5.5ml/min. LC-MS method in using system ID is water’s UPLC H-class with PDA and SQD mass detector. The describe LC-MS method was linear over a concentration range of MET, DFZ, EFZ and CFZ were found to be 5-30μg/ml, 1-5 μg/ml, 2-10μg/ml and 5-30μg/ml respectively. The retention times of MET, DFZ, EFZ and CFZ were found to be 1.16, 1.25, 1.39 and 1.83min respectively theoretical plate count as per ICH limits. The limit of detection is achieved on lowest level for making sensitive and robust method for simultaneous assay analysis. The LC-MS method was validated and was successfully employed for the analysis of pharmaceutical formulation containing Metformin, Dapagliflozin, Empagliflozin and Canagliflozin in combined tablet dosage form.

Keywords: Antidiabetic drugs: Metformin, Canagliflozin, Empagliflozin, Dapagliflozin, LC–MS, Development and Validation.


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