Department of Legal Medicine, Forensic and Pharmaco-Toxicological Science, University of Pavia, 27100 Pavia, Italy.
Anal Biochem. 2011 Nov 1;418(1):30-6. doi: 10.1016/j.ab.2011.06.038. Epub 2011 Jul 5.
The aim of this study was to develop a method for the determination of ethyl-glucuronide (EtG) and ethyl-sulfate (EtS), two direct ethanol metabolites, in early placental and fetal human tissues, as potential biomarkers of transplacental ethanol transfer from the mother to the fetus. Placental and fetal tissue samples were obtained from women undergoing voluntary termination of pregnancy at 12 weeks of gestation. Samples were deproteinized and directly injected into a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) system. Limits of detection of 13.0 and 23.0 pmol/g and lower limits of quantification of 22.0 and 40.0 pmol/g were reached for EtG and EtS, respectively. Inter- and intraday imprecision and accuracy were always lower than 15%. The method was applied to 70 samples (35 placentas and 35 fetal tissues). Of 35 samples, 4 samples collected from 4 women tested positive for EtG and EtS, always showing higher concentrations for EtG. The placenta/fetal tissue ratio for EtG was 2.9 ± 0.9, whereas EtS showed a ratio of 1.7 ± 0.7. Preliminary results suggest that these metabolites are present in both tissues. Further studies should now corroborate the hypothesis, not yet confirmed, that transplacental transfer of ethanol takes place not only for the parent compound but also for EtG and EtS.
本研究旨在建立一种测定胎盘和胎儿人组织中两种直接乙醇代谢物乙基-葡萄糖醛酸苷（EtG）和乙基硫酸盐（EtS）的方法，作为从母体向胎儿转移的潜在生物标志物。在妊娠 12 周时，从自愿终止妊娠的女性中获取胎盘和胎儿组织样本。将样本进行蛋白沉淀，直接注入液相色谱-串联质谱（LC-MS/MS）系统。对于 EtG 和 EtS，检测限分别达到 13.0 和 23.0 pmol/g，定量下限分别达到 22.0 和 40.0 pmol/g。日内和日间精密度和准确度始终低于 15%。该方法应用于 70 个样本（35 个胎盘和 35 个胎儿组织）。在 35 个样本中，有 4 个样本来自 4 名女性，检测到 EtG 和 EtS 阳性，且 EtG 的浓度总是更高。对于 EtG，胎盘/胎儿组织比值为 2.9 ± 0.9，而 EtS 显示的比值为 1.7 ± 0.7。初步结果表明，这些代谢物存在于两种组织中。现在应该进行进一步的研究来证实尚未证实的假设，即乙醇不仅通过母体化合物而且通过 EtG 和 EtS 进行胎盘转移。
