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Development and validation of a liquid chromatographic/electrospray ionization mass spectrometric method for the determination of benazepril, benazeprilat and hydrochlorothiazide in human plasma
Author(s) -
Vonaparti Ariadni,
Kazanis Michael,
Panderi Irene
Publication year - 2006
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.1014
Subject(s) - chemistry , chromatography , electrospray ionization , detection limit , mass spectrometry , benazepril , high performance liquid chromatography , hydrochlorothiazide , analytical chemistry (journal) , medicine , blood pressure , radiology
A new method was developed and fully validated for the quantitation of benazepril, benazeprilat and hydrochlorothiazide in human plasma. Sample pretreatment was achieved by solid‐phase extraction (SPE) using Oasis HLB cartridges. The extracts were analysed by high‐performance liquid chromatography (HPLC) coupled to a single‐quadrupole mass spectrometer (MS) with an electrospray ionization interface. The MS system was operated in selected ion monitoring (SIM) modes. HPLC was performed isocratically on a reversed‐phase porous graphitized carbon (PGC) analytical column (2.1 × 125.0 mm i.d., particle size 5 µm). The mobile phase consisted of 55% acetonitrile in water containing 0.3% v/v formic acid and pumped at a flow rate of 0.15 ml min −1 . Chlorthalidone was used as the internal standard (IS) for quantitation. The assay was linear over a concentration range of 5.0–500 ng ml −1 for all the compounds analysed, with a limit of quantitation of 5 ng ml −1 for all the compounds. Quality control (QC) samples (5, 10, 100 and 500 ng ml −1 ) in five replicates from three different runs of analyses demonstrated intra‐assay precision (coefficient of variation (CV) ≤14.6%), inter‐assay precision (CV ≤ 5.6%) and overall accuracy (relative error less than −8.0%). The method can be used to quantify benazepril, benazeprilat and hydrochlorothiazide in human plasma, covering a variety of pharmacokinetic or bioequivalence studies. Copyright © 2006 John Wiley & Sons, Ltd.