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J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Apr 15;957:110-5. doi: 10.1016/j.jchromb.2014.03.002. Epub 2014 Mar 13.

Simultaneous determination of irbesartan and hydrochlorothiazide in human plasma by ultra high performance liquid chromatography tandem mass spectrometry and its application to a bioequivalence study.

Author information

  • 1Medical College of Henan University of Science and Technology, Luoyang 471003, China.
  • 2The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
  • 3The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
  • 4The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China. Electronic address: xurenai1986@163.com.

Abstract

An ultra high performance liquid chromatography tandem mass spectrometry (U-HPLC-MS/MS) method was developed and validated to determine irbesartan (IRB) and hydrochlorothiazide (HCTZ) in human plasma simultaneously. Plasma samples were prepared using protein precipitation with acetonitrile, the two analytes and the internal standard losartan were separated on an Acquity U-HPLC BEH C18 column and mass spectrometric analysis was performed using a QTrap5500 mass spectrometer coupled with an electro-spray ionization (ESI) source in the negative ion mode. The MRM transitions of m/z 427.2→206.9 and m/z 296.1→204.9 were used to quantify for IRB and HCTZ, respectively. The linearity of this method was found to be within the concentration range of 5-3000ng/mL for IRB, and 0.5-300ng/mL for HCTZ in human plasma, respectively. The lower limit of quantification (LLOQ) was 5ng/mL and 0.5ng/mL for IRB and HCTZ in human plasma, respectively. The relative standard deviations (RSD) of intra and inter precision were less than 12% for both IRB and HCTZ. The analysis time of per sample was 2.5min. The developed and validated method was successfully applied to a bioequivalence study of IRB (300mg) with HCTZ (12.5mg) tablet in Chinese healthy volunteers (N=20).

Copyright © 2014 Elsevier B.V. All rights reserved.

KEYWORDS:

Bioequivalence; Human plasma; Hydrochlorothiazide; Irbesartan; U-HPLC–MS/MS

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