Janda Ines, Weinmann Wolfgang, Kuehnle Thorsten, Lahode Martina, Alt Andreas
Department of Forensic Medicine, University Ulm, Prittwitzstr. 6, D-89075 Ulm, Germany.
Forensic Sci Int. 2002 Aug 14;128(1-2):59-65. doi: 10.1016/s0379-0738(02)00163-9.
A method for the sensitive and selective determination of ethyl glucuronide (EtG) in hair has been developed using solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Washed and cut hair segments were extracted by ultrasonication (3h, 50 degrees C) and the extracts were cleaned-up with aminopropyl SPE columns. LC-MS/MS analysis was performed using a polar-endcapped phenyl-hexyl-RP-phase with negative mode electrospray ionisation (ESI) using a triple quadrupole mass spectrometer (Sciex API 365) with a turboionspray source and post-column addition of acetonitrile for enhanced sensitivity. The MS/MS transitions monitored were m/z 221 -->75 for EtG and 226 -->75 for D(5)-EtG as an internal standard. The method was selective and sensitive, with a detection limit of 51 pg/mg hair at a signal-to-noise ratio of 3:1. The mean recovery was 96%, with an intra- and inter-day precision of less than 11.7% at a concentration of 200 pg/mg. The linearity was assessed in the range of 25-2000 pg/mg hair, with a correlation coefficient of 0.997. The method was successfully applied to 97 human hair samples which were taken at autopsies from persons with known alcoholism or were obtained from alcoholics who were hospitalized for ethanol withdrawal, from social drinkers and from children having not consumed any alcohol. Although, approximately two-third of the alcoholics showed EtG concentrations in hair of higher than 51 pg/mg (up to >4000 pg/mg), in one-third the EtG concentration was below the detection limit. However, only in one of five hair samples of "social drinkers", the EtG concentration was above the detection limit (51 pg/mg). No EtG has been detected in the hair of children. These investigations demonstrate that heavy alcohol consumption may be but not necessarily has to be detectable by EtG analysis in hair.
已开发出一种使用固相萃取（SPE）和液相色谱 - 串联质谱（LC - MS/MS）灵敏且选择性地测定毛发中葡萄糖醛酸乙酯（EtG）的方法。将洗净并剪好的毛发段通过超声处理（3小时，50摄氏度）进行提取，提取物用氨丙基SPE柱进行净化。使用带有涡轮离子喷雾源的三重四极杆质谱仪（Sciex API 365），在负极封端的苯基 - 己基反相柱上，采用负模式电喷雾电离（ESI）进行LC - MS/MS分析，并在柱后添加乙腈以提高灵敏度。监测的MS/MS跃迁对于EtG为m/z 221→75，对于作为内标的D(5)-EtG为m/z 226→75。该方法具有选择性和灵敏性，在信噪比为3:1时，检测限为51 pg/mg毛发。平均回收率为96%，在浓度为200 pg/mg时，日内和日间精密度均小于11.7%。线性范围评估为25 - 2000 pg/mg毛发，相关系数为0.997。该方法成功应用于97份人类毛发样本，这些样本取自已知酗酒者的尸检，或从因乙醇戒断住院的酗酒者、社交饮酒者以及未饮用过任何酒精的儿童处获得。尽管约三分之二的酗酒者毛发中的EtG浓度高于51 pg/mg（高达>4000 pg/mg），但三分之一的酗酒者EtG浓度低于检测限。然而，在“社交饮酒者”的五份毛发样本中，只有一份的EtG浓度高于检测限（51 pg/mg）。在儿童毛发中未检测到EtG。这些研究表明，大量饮酒可能但不一定能通过毛发中EtG分析检测到。
