Reverse-phase high performance liquid chromatography for the determination of tiopronin in human plasma after derivatization with p-bromophenacyl bromide

In the study, we developed a simple, rapid and sensitive method for the determination of tiopronin (TP) in human plasma, which was based on derivatization with p-bromophenacyl bromide ( p-BPB) followed by liquid–liquid extraction and reverse-phase HPLC-UV detection. For the first time, the p-BPB was...

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Bibliographic Details
Published in:Analytica chimica acta Vol. 565; no. 2; pp. 178 - 182
Main Authors: Huang, Taomin, Yang, Bei, Yu, Yingjia, Zheng, Xiaowei, Duan, Gengli
Format: Journal Article
Language:English
Published: Amsterdam Elsevier B.V 21-04-2006
Elsevier
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Summary:In the study, we developed a simple, rapid and sensitive method for the determination of tiopronin (TP) in human plasma, which was based on derivatization with p-bromophenacyl bromide ( p-BPB) followed by liquid–liquid extraction and reverse-phase HPLC-UV detection. For the first time, the p-BPB was introduced into the derivatization of TP. The thiol group of TP was trapped with p-BPB to form a TP- p-BPB adduct, which can be very suitable for UV detection. From acidified plasma samples, the derivatized TP was extracted with 5 mL dichloromethane. Effective chromatographic separation was achieved using a C18 column (DIAMONSIL 150 mm × 4 mm i.d., 5 μm) based on an acetonitrile–water–trifluoroacetic acid (40:59.88:0.12, v/v/v) elution at a flow-rate of 1 mL/min. The IS and the derivatized TP were detected at 263 nm. No endogenous substances were found to interfere. The limit of quantification for derivatized TP (TP- p-BPB) in plasma was 40 ng/mL. The calibration curve for the derivatized TP showed linearity in the range 0.04–4 μg/mL with a regression coefficient corresponding to 0.9991 and the coefficient of the variation of the points of the calibration curve being lower than 10%. Extraction recoveries of the derivatized TP in plasma were greater than 72%. The method was suitably validated and successfully applied to determination of TP in human plasma samples.
ISSN:0003-2670
1873-4324
DOI:10.1016/j.aca.2006.02.049