Bioanalytical Stability Indicating Method Development and Validation for the Estimation of Anti Diabetic Drugs in Human Plasma by Using RP –HPLC Method.

Abstract

A reverse phase HPLC has been developed and validated for estimation of Metformin and Dapagliflozin in Human plasma. In this method, a reverse phase column Agilent – C18 Plasma 5 μm (4.6× 250 mm) as a Stationary phase, with a Mobile Phase of Methanol and 0.05% ortho phosphoric acid in water (pH 2.5) in the ratio of (50%:50%) at 0.8 mL/min flow rate was used with a DAD detection was carried out at 233nm. The volume injected was 20 µL. The described HPLC method was linear over a concentration range of 100 to 500 µg/ml for Metformin and 2 to 10µg/ml for Dapagliflozin. The Metformin showed retention time 2.222 min &Dapagliflozin showed retention time 4.429 min respectively. The Accuracy and Precision, recovery, selectivity for metformin was 100-500 µg/ml respectively& for Dapagliflozin 2 to 10  µg/ml. The stability of the drug spiked human during three freeze thaw cycles were stable in plasma for about one month when stored at frozen state.