Original Paper

Analytical and Bioanalytical Chemistry

, Volume 382, Issue 4, pp 1049-1054

Development and validation of a selective and robust LC-MS/MS method for quantifying amlodipine in human plasma

  • P. MassarotiAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , L. A. B. MoraesAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , M. A. M. MarchiorettoAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , N. M. CassianoAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , G. BernasconiAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , S. A. CalafattiAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , F. A. P. BarrosAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School
  • , E. C. MeurerAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School Email author 
  • , J. PedrazzoliAffiliated withClinical Pharmacology and Gastroenterology Unit, São Francisco University Medical School

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Abstract

A liquid chromatographic–tandem mass spectrometric method (LC-MS/MS) for quantifying amlodipine in human plasma was developed and validated. Sample preparation was based on liquid–liquid extraction using NaOH and a mixture of ethyl acetate/hexane (80/20; v/v). Chromatography was performed on a C-18 analytical column and the retention times were 1.9 and 3.0 min for amlodipine and nimodipine (internal standard), respectively. The ionization was optimized using ESI(+) and enhanced selectivity was achieved using tandem mass spectrometric analysis via two MRM functions, 409→238 and 418→343 for amlodipine and nimodipine. The calibration curve ranged from 0.2 to 20.0 ng/mL. The inter-day precision and accuracy and the relative standard deviation (RSD) were <15%. The analyte was shown to be stable over the timescale of the whole procedure. The robustness of the method was demonstrated by the good reproducibility of the results obtained during the analysis of clinical samples.

Keywords

Mass spectrometry Bioequivalence Amlodipine