Hegstad Solfrid, Helland Arne, Hagemann Cecilie, Spigset Olav
Department of Clinical Pharmacology, St. Olav University Hospital, 7006 Trondheim, Norway.
Department for Laboratory Medicine, Children's and Women's Health, Norwegian University of Science and Technology, 7491 Trondheim, Norway.
J Anal Toxicol. 2017 Sep 1;41(7):618-622. doi: 10.1093/jat/bkx032.
A method including semi-automated extraction of ethyl glucuronide (EtG) and ethyl sulfate (EtS) from serum followed by ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS-MS) has been developed and validated. Sample preparation prior to UHPLC-MS-MS analysis consisted of protein precipitation and filtration through a phospholipid removal plate. Chromatography was achieved using an HSS T3 column and gradient elution with formic acid in water in combination with methanol. The mass spectrometer was monitored in the negative mode with multiple reaction monitoring. Two transitions were monitored for the analytes and one for the deuterated internal standards (ISs). The limits of quantification were 0.025 mg/L for EtG and 0.009 mg/L for EtS. The between-assay relative standard deviations were in the range of 3.8-9.1%, the recovery was 66-102% and matrix effects ranged from 88 to 97% (corrected with IS). Compared to previously published studies, the method presented is semi-automated, uses a simple method for phospholipid removal and has short run times and low limit of quantifications. We analyzed serum samples from 49 female patients presenting to the Sexual Assault Centre at St. Olav University Hospital in Trondheim, Norway, for ethanol, EtG and EtS. EtG and EtS were detected longer than ethanol itself after intake of ethanol, with estimated maximum detection times of >24 h. The ethanol, EtG and EtS concentrations were highly correlated (P < 0.001), but with large inter-individual variations. This study suggests that analysis of EtG and EtS in serum or blood may complement ethanol analysis and shed light on the patient's recent ethanol intake after ethanol itself is no longer detectable.
已开发并验证了一种从血清中半自动提取葡萄糖醛酸乙酯（EtG）和硫酸乙酯（EtS），随后进行超高效液相色谱 - 串联质谱法（UHPLC - MS - MS）分析的方法。UHPLC - MS - MS分析前的样品制备包括蛋白质沉淀和通过磷脂去除板过滤。使用HSS T3柱进行色谱分离，并采用甲酸与水混合甲醇的梯度洗脱。质谱仪在负离子模式下通过多反应监测进行监测。对分析物监测两个跃迁，对氘代内标（ISs）监测一个跃迁。EtG的定量限为0.025 mg/L，EtS的定量限为0.009 mg/L。批间相对标准偏差在3.8 - 9.1%范围内，回收率为66 - 102%，基质效应在88%至97%之间（用内标校正）。与先前发表的研究相比，所提出的方法是半自动的，采用简单的磷脂去除方法，运行时间短且定量限低。我们分析了挪威特隆赫姆市圣奥拉夫大学医院性侵犯中心49名女性患者血清样本中的乙醇、EtG和EtS。摄入乙醇后，EtG和EtS的检测时间比乙醇本身更长，估计最大检测时间>24小时。乙醇、EtG和EtS的浓度高度相关（P < 0.001），但个体间差异较大。这项研究表明，血清或血液中EtG和EtS的分析可能补充乙醇分析，并在乙醇本身不再可检测后揭示患者近期的乙醇摄入量。
