Liquid chromatography–mass spectrometry method for determination of tetramethylpyrazine and its metabolite in dog plasma
- 作者：Wang ; Peng ; Jin ; Xin ; Qi ; Meiling ; Fang ; Lin
- 关键词：Tetramethylpyrazine ; Metabolite ; Liquid chromatography&ndash ; mass spectrometry ; Dog plasma
- 刊名：Journal of Chromatography B ; Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2004
- 期刊代码：124_15700232
- 类别：ch
- 出版时间：December 25, 2004
- 卷：813
- 期：1-2
- 页码：263-268
- 文件大小：128 K
摘要
A liquid chromatography–mass spectrometry method is described for the determination of tetramethylpyrazine (TMP) and its active metabolite, 2-hydroxymethyl-3,5,6-trimethylpyrazine (HTMP) in dog plasma. This method involves a plasma clean-up step using protein precipitation procedure followed by LC separation and positive electrospray ionization mass spectrometry detection (ESI-MS). Chromatographic separation of the analytes was achieved on a C18 column using a mobile phase of methanol, water and acetic acid (50:50:0.6, v/v/v) at a flow rate of 1.0ml/min. Selected ion monitoring (SIM) mode was used for analyte quantitation at m/z 137.2 for TMP, m/z 153.2 for HTMP and m/z 195.2 for caffeine. The linearity was obtained over the concentration ranges of 20–6000ng/ml for TMP and 20–4000ng/ml for HTMP and the lower limit of quantitation was 20ng/ml for both analytes. For each level of QC samples, both inter- and intra-day precisions (R.S.D.) were ≤7.4%for TMP and ≤6.0%for HTMP, and accuracy (R.E.) was ±6.0%for TMP and ≤3.5%for HTMP. The proposed LC–MS method was successfully applied to the pharmacokinetic studies of a TMP formulation preparation after oral administration to beagle dogs.