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Validation of a HPLC‐ESI MS/MS method for the determination of clonidine in human plasma and its application in a bioequivalence study in Chinese healthy volunteers
Author(s) -
Zhuang Jialang,
Chen Jiangying,
Wang Xueding,
Pang Yin,
Bi Huichang,
Huang Lihui,
Zeng Guixiong,
Liao Xiaoxing,
Ma Zhongfu,
Chen Xiao,
Zhong Guoping,
Huang Min,
Zhao Xianglan
Publication year - 2015
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.3450
Subject(s) - chromatography , chemistry , bioequivalence , electrospray ionization , mass spectrometry , selected reaction monitoring , formic acid , extraction (chemistry) , high performance liquid chromatography , detection limit , sample preparation , tandem mass spectrometry , pharmacokinetics , pharmacology , medicine
A rapid and sensitive liquid chromatography–tandem mass spectrometry (HPLC‐MS/MS) method to determine clonidine in human plasma was developed and fully validated. Sample preparation was involved an one‐step extraction with diethyl ether. Donepezil was employed as the internal standard (IS). Chromatographic separation was performed on a Hypersil BDS C 18 column (i.d. 2.1 × 50 mm, particle size 3μm) with a mobile phase of methanol–water (containing 0.1% formic acid; 60:40, v/v) at a flow rate of 200 μL/min. The peaks were detected by mass spectrometry using the electrospray ion source in selected reaction monitoring mode. The extraction recovery was 72.53–85.25%. The method was found to be linear in a concentration range of 0.02–6.00 ng/mL and the lower limit of quantification was 0.02 ng/mL. The within‐ and between‐batch precisions at three concentrations were 4.33–16.47 and 7.24–17.24% with accuracies of −2.47–10.91 and 1.86–10.19%, respectively. This validated method was successfully used for a bioequivalence study of two clonidine transdermal patches on healthy volunteers. The results suggested that the test formulation of clonidine patch met the regulatory criterion for bioequivalence to the reference formulation, but a larger sample size should be needed for the estimation of bioequivalence. Copyright © 2015 John Wiley & Sons, Ltd.