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Highly sensitive and rapid ultra‐performance liquid chromatography–tandem mass spectrometry method for the determination of nifedipine in human plasma and its application to a bioequivalence study
Author(s) -
Patel Daxesh P.,
Sharma Primal,
Sanyal Mallika,
Singhal Puran,
Shrivastav Pranav S.
Publication year - 2012
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.2725
Subject(s) - chemistry , chromatography , selected reaction monitoring , bioequivalence , mass spectrometry , extraction (chemistry) , high performance liquid chromatography , analyte , triple quadrupole mass spectrometer , ammonium acetate , tandem mass spectrometry , liquid chromatography–mass spectrometry , solid phase extraction , nifedipine , analytical chemistry (journal) , pharmacokinetics , calcium , medicine , organic chemistry
An ultra performance liquid chromatography–tandem mass spectrometry (UPLC‐MS/MS) method has been developed for the determination of nifedipine in human plasma using nifedipine‐d6 as the internal standard (IS). The plasma samples were prepared by solid‐phase extraction on Phenomenex Strata‐X cartridges employing 200 μL human plasma. Chromatography was carried out on Waters Acquity UPLC BEH C 18 (50 × 2.1 mm, 1.7 µm particle size) analytical column under isocratic conditions using a mobile phase consisting of 4.0 m m ammonium acetate‐acetonitrile (15:85, v/v). The precursor → product ion transitions for nifedipine ( m/z 347.2 → 315.2) and IS ( m/z 353.1 → 318.1) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring and positive‐ion mode. The method was validated over a wide dynamic concentration range of 0.050–150 ng/mL. Matrix effect was assessed by post‐column analyte infusion and the mean extraction recovery was 95.6% across four quality control levels. The method is rugged and rapid with a total run time of 1.2 min and was applied to a bioequivalence study of 20 mg nifedipine tablet formulation in 30 healthy Indian subjects under fasting condition. Assay reproducibility was confirmed by reanalysis of 116 incurred samples. Copyright © 2012 John Wiley & Sons, Ltd.