Institute of Forensic Medicine, Forensic Toxicology, Freiburg University Medical Centre, Albertstrasse 9, 79104 Freiburg, Germany.
Anal Bioanal Chem. 2009 Dec;395(8):2499-507. doi: 10.1007/s00216-009-3036-x.
A new quantitation method for the determination of drugs of abuse (opiates, amphetamine and derivatives, cocaine, methadone and metabolites) in serum by using online extraction coupled to liquid chromatography (LC)-mass spectrometry (MS)/MS has been developed. The online extraction is carried out using two extraction columns simultaneously and one analytical column. One extraction column is loaded, while the other one is eluted by a gradient. The elution gradient also separates the analytes in the analytical column. For the sample preparation, serum is spiked with a mixture of deuterated analogues of the drugs. After protein precipitation with methanol/zinc sulphate, centrifugation, evaporation and reconstitution, the sample is injected into the LC system. The quantitation is based on the analysis of two multiple reaction monitoring transitions per drug. The recovery of the protein precipitation step is over 80% for all analytes. Intra- and interday precision, as relative standard deviation, is lower than 6%, and in the case of accuracy, RE is lower than 15%. Only the most polar analytes showed matrix effects. The limits of quantitation for the analysed compounds vary between 0.5 and 2.8 ng/mL. The developed method was used to quantify basic drugs in samples "from driving under the influence of drugs" cases. The results were compared with those obtained by using solid-phase extraction-GC-MS.
一种新的定量分析方法，用于通过在线提取结合液相色谱（LC）-质谱（MS）/MS 来检测血清中的滥用药物（阿片类药物、苯丙胺及其衍生物、可卡因、美沙酮及其代谢物）。在线提取同时使用两个萃取柱和一个分析柱进行。一个萃取柱加载，另一个用梯度洗脱。洗脱梯度还分离分析柱中的分析物。对于样品制备，血清用药物的氘代类似物混合物进行标记。用甲醇/硫酸锌沉淀蛋白后，离心、蒸发和复溶，将样品注入 LC 系统。定量基于每个药物的两个多重反应监测转换的分析。所有分析物的蛋白沉淀步骤回收率均高于 80%。日内和日间精密度（以相对标准偏差表示）低于 6%，准确度（RE）低于 15%。只有最极性的分析物显示出基质效应。分析化合物的定量限在 0.5 至 2.8ng/mL 之间变化。所开发的方法用于定量“药物影响驾驶”案例中基本药物的样本。将结果与固相萃取-GC-MS 获得的结果进行比较。
