Selective detection of phosphatidylethanol homologues in blood as biomarkers for alcohol consumption by LC-ESI-MS/MS
Selective detection of phosphatidylethanol homologues in blood as biomarkers for alcohol consumption by LC-ESI-MS/MS
复制标题
DOI：
10.1002/jms.1608
复制
发表时间：
2009-09-01
影响因子：
2.3
通讯作者：
Bouzas, N. Ferreiros
中科院分区：
文献类型：
--
作者：
Gnann, H.;Weinmann, W.;Bouzas, N. Ferreiros
采用LC-ESI-MS/MS，以磷脂酰丁醇（PBut）为内标，采用小型有机溶剂萃取和反相色谱法，建立了一种新的定量检测异常磷脂-磷脂酰乙醇（PEth）的方法。研究人员在尸体解剖时从酗酒者身上采集的死后血液中，以及在一名志愿者的全血样本中，检测了具有两种脂肪酸取代基的PEth同源物——PEth 18:1/ 18:1， PEth 16:0/ 16:0:0。此外，还检测到PEth 18:1/16:0或其等压异构体PEth - 16:0/ 18:1。与以往的高效液相色谱（HPLC）蒸发光散射检测（ELSD）方法相比，LC-MS/ ms方法灵敏度更高，检测限低于20 ng/ml，对单一PEth同源物的选择性更强，而ELSD用于检测PEth同源物的总和，灵敏度低约10倍。LC-MS/MS可以监测PEth同源物作为有害和长时间饮酒的生物标志物，就像早期的HPLC/ELSD一样，只有在每天摄入超过50克乙醇超过2周后，血液中才能测量PEth。由于其较高的灵敏度，当苯醚形成的浓度很低时，有可能通过LC-MS/MS检测到单次重度饮酒。这开辟了PEth应用的新领域，以较低的频率发现单次或多次重度饮酒，并且与HPLC/ELSD或使用其他直接标记物（如葡萄糖醛酸乙酯或硫酸乙酯）相比，在血液中具有更大的检测窗口。版权所有John Wiley & Sons， Ltd. 2009
A new validated method for the quantitation of the abnormal phospholipid phosphatidylethanol (PEth) - a biomarker for ethanol uptake - has been developed by LC-ESI-MS/MS following miniaturised organic solvent extraction and reversed phase chromatography with phosphatidylbutanol (PBut) as internal standard. PEth homologues with two fatty acid substituents - PEth 18:1/18: 1, PEth 16:0/16: 0 - were determined in post-mortem blood collected from heavy drinkers at autopsy and also in whole blood samples from a volunteer after a single 60 g-dose of ethanol. Furthermore, PEth 18:1/16:0 or its isobaric isomer PEth - 16:0/18: 1 was detected. In comparison to previous high-performance liquid chromatography (HPLC) methods with evaporative light scattering detection (ELSD), the LC-MS/MS-method is more sensitive - with a limit of detection below 20 ng/ml - and more selective for single PEth homologues, while ELSD has been used for detection of the sum of PEth homologues with approximately 10 times less sensitivity. LC-MS/MS enables monitoring of PEth homologues as biomarkers for harmful and prolonged alcohol consumption as with HPLC/ELSD earlier, where PEth is measurable in blood only after more than 50 g ethanol daily intake for more than 2 weeks. Because of its higher sensitivity, there is a potential to detect single heavy drinking by LC-MS/MS, when PEth is formed in very low concentrations. This opens a new field of application of PEth to uncover single or multiple heavy drinking at a lower frequency and with a larger window of detection in blood than before by HPLC/ELSD or by use of other direct markers, e.g. ethyl glucuronide or ethyl sulfate. Copyright (C) 2009 John Wiley & Sons, Ltd.