3RD INTERNATIONAL CONGRESS ON TECHNOLOGY - ENGINEERING & SCIENCE - Kuala Lumpur - Malaysia (2017-02-09)

Simultaneous Determination Of Ramipril, Glimepiride And Metformin In Human Plasma By Ultra Performance Liquid Chromatography – Tandem Mass Spectrometry

Ramipril (RAM), glimepiride (GLM) and metformin (MET) are recently formulated together for the treatment of diabetes and cardiovascular disorders. This study introduces a liquid chromatography–tandem mass spectrometric (LC-MS/MS) method for the simultaneous quantitative determination of the three drugs in human plasma after liquid-liquid extraction. This method used of electrospray ionization in positive mode for all analytes using triple quadrupole mass spectrometry where atorvastatin was used as internal standard (IS). Analytes were extracted from plasma by n-hexane: butanol (50:50%, v/v) subsequently separated on Waters AcquityTM UPLC BEH shield RP-C18 column (150 mm x 2.1 mm, 1.7 µm) using methanol: water containing 0.1% formic acid (90: 10%, v/v) as a mobile phase, at a flow rate of 0.2 mL/min. Quantification of RAM, GLM, MET and IS was performed using multi-reaction monitoring mode (MRM). The calibration curves were linear (r2 > 0.99) over the concentration range of 0.1- 1000 ng/mL for RAM & GLM and 250- 2000 ng/mL for MET. The intra-day and inter-day precisions were below 14.32% and the accuracy was all within ±15%. Moreover, other validation parameters for the proposed method such as matrix effect, selectivity, recovery and stability were adopted. In conclusion, the outcomes of the present study showed that, it can be used for the estimation of the three drugs for bioavailability and pharmacokinetics studies.
Eman Elzanfaly, Sherif Abdel-Gawad