[ Pharmaceutical Sciences Asia - ONLINE ]
E-ISSN 2586-8470
[ Journal Abbreviation: Pharm.Sci.Asia ]
Mahidol University Journal of Pharmaceutical Sciences
  FORMER NAME   "Mahidol University Journal of Pharmaceutical Sciences" Published Since 1974


DOI: https://doi.org/10.29090/psa.2017.04.217Pharm Sci Asia 2017; 44(4), 217-226

Development and Validation of Liquid Chromatography Tandem Mass Spectrometry Method for Determination of Amlodipine in Human Plasma and its Application

P. Pongnarin, W. Thanakosai, P. Kingkhangphloo, W. Chandranipapongse, S. Kongpatanakul, S. Chatsiricharoenkul*

- Department of Pharmacology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

Asensitive liquid chromatography tandem mass spectrometry method was developed to quantify amlodipinein human plasma. Alkalinized plasma spiked with desipramine, an internal standard, was extracted by liquid-liquid extraction andevaporated an organic part to dryness. The residue was reconstituted and injected into anAcquityUltra Performance LCTM, (Waters, Co., Ltd.USA) with C18 column. The isocratic elution of mobile phasewas performed by 90% of acetonitrile and 10% of 10 mM ammonium acetate pH 4 atflow rate of 0.20 mL/min with 5 minutes of total run time. Mass spectrometric analysis was performed using a Quattro Premier XE mass spectrometer, (Micromass Technologies, UK) coupled with an electrospray ionization (ESI) source in the positive ion mode. The MRM transitions of m/z 409.17>238.19 and 409.17>294.09were selected for amlodipineand 267.09>208.06for desipramine. The retention times were 1.63 and 1.69 minutes for amlodipineand desipramine, respectively. The linearity of the method revealed a correlation coefficient of >0.998 within the concentration range of 0.05 - 20 ng/mL. This work has been fully validated according to the Guidance for Industry: Bioanalytical Method Validation (USFDA CDER, 2001, BP) with high degree of accuracy and precision. This assay method was applied to quantify amlodipine concentrations in human plasma samples in a bioequivalence study.


amlodipine, LC-MS/MS, Method development, Method Validation

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