HPLC-MS/MS methods for the quantitative analysis of ophthalmic acid in rodent plasma and hepatic cell line culture medium
HPLC-MS/MS methods for the quantitative analysis of ophthalmic acid in rodent plasma and hepatic cell line culture medium
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DOI：
10.1016/j.jpba.2010.11.038
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发表时间：
2011-04-05
影响因子：
3.4
通讯作者：
Wilson, Ian
中科院分区：
文献类型：
--
作者：
Geenen, Suzanne;Michopoulos, Filippos;Wilson, Ian
眼酸是谷胱甘肽的内源性三肽类似物，已被认为是扑热息痛/对乙酰氨基酚肝毒性的潜在生物标志物。本研究建立了高效液相色谱-串联质谱法，对体外细胞培养上清液和血浆中的OA进行准确、灵敏、特异的检测和定量。对于细胞培养液，最低检出限为1 ng/ml，所有浓度的样品携带之间的精密度小于1%，日内精密度低于15%，日间分析精密度低于9%。对于大鼠血浆，内源性OA的存在导致LLOQ为25 ng/ml(定义为校准曲线上的最低浓度，其中基峰小于LLOQ的20%)。对于血浆分析，所有浓度的残留率小于1%，批内和批间精密度低于21%。该方法对两种样品的线性范围均为5微克/毫升。该方法已成功地应用于慢性给药大鼠肝毒素甲基吡喃后的样品中，血浆中的OA浓度与已建立的肝损伤生物标志物如天冬氨酸氨基转移酶(AST)的浓度呈弱负相关。(C)2010爱思唯尔B.V.保留所有权利。
Ophthalmic acid (OA), an endogenous tripeptide analogue of glutathione, has been suggested as a potential biomarker for paracetamol/acetaminophen hepatotoxicity. Here HPLC-MS/MS methods have been developed for the precise, sensitive and specific detection and quantification of OA in in vitro cell culture medium and plasma. For the cell culture medium the LLOQ was found to be 1 ng/ml, with less than 1% between sample carry over at all concentrations and precision below 15% for within day and below 9% for between day analyses. For rat plasma the presence of endogenous OA resulted in the LLOQ being 25 ng/ml (defined as the lowest concentration on the calibration curve where the base peak was less than 20% of the LLOQ). For the plasma assay the percentage carry over was less than 1% for all concentrations and within and between batch precision was below 21%. The methods were linear for both sample types from the LLOQ up to 5 mu g/ml. The method was successfully applied to the determination of OA in samples obtained following the chronic administration of the rat hepatotoxin methapyrilene, where plasma OA concentrations were observed to show a weak negative correlation with those of established liver injury biomarkers such as aspartate aminotransferase (AST). (C) 2010 Elsevier B.V. All rights reserved.