Abstract

DETERMINATION OF NIFEDIPINE IN HUMAN PLASMA BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY AND ITS VALIDATION

Srinivasa Rao Baratam*, Kalyan Chakravarthy Janjanam and Vijaya Ratna J.

ABSTRACT

A selective, rapid and sensitive high-performance liquid chromatography–tandem mass spectrometry (HPLC -MS/MS) method was developed and validated for the quantification of nifedipine in human plasma. Nimodipine was used as an internal standard, sample pretreatment involved a simple solid phase extraction using HLB Cartridge. The analysis was carried out on a Hypersil, gold C18 column (50 mm×4.6 mm, 5μ) with flow rate of 0.6 mL/min. The mobile phase was consisting of 0.1% formic acid buffer and acetonitrile in the ratio of 30:70 v/v. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode. Linear calibration curves were obtained in the concentration range of 0.5 to 100 ng/mL. The intra- and inter-day precision (R.S.D.) and accuracy results were within the accepted limits as per USFDA guidelines.

Keywords: Nifedipine; Nimodipine; Solid–phase extraction (SPE); LC–MS/MS; Multiple reaction monitoring (MRM) mode; Method validation.