Institute of Legal Medicine, University Hospital Charité, Hittorfstr. 18, 14195 Berlin, Germany.
Anal Bioanal Chem. 2010 Apr;396(7):2469-77. doi: 10.1007/s00216-010-3474-5. Epub 2010 Feb 10.
Fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) were determined in 602 meconium samples in a maternal health evaluation study for detection of gestational alcohol consumption. A validated headspace solid phase microextraction method in combination with GC-MS was used for FAEE and the cumulative concentration of ethyl palmitate, ethyl linoleate, ethyl oleate, and ethyl stearate with a cut-off of 500 ng/g was applied for interpretation. A new and simple method was developed and validated for quantification of EtG from 10-20 mg meconium with D(5)-EtG as internal standard consisting of 30 min. extraction with methanol/water (1:1, v/v), evaporation of methanol, filtration of the aqueous solution through a cellulose filter and injection into LC-MS-MS. The limits of detection and quantification for EtG were 10 and 30 ng/g, the recovery 86.6 to 106.4% and the standard deviation of the concentrations ranged from 13% at 37 ng/g to 5% at 46,700 ng/g (N = 6). FAEE above the cut-off were found in 43 cases (7.1%) with cumulative concentrations between 507 and 22,580 ng/g and with one outlier of about 150,000 ng/g (EtG not detected). EtG was detected in 97 cases (16.3%) and concentrations between LOD and 10,200 ng/g with another outlier of 82,000 ng/g (FAEE 10,500 ng/g). Optimal agreement between the two markers was obtained with a cut-off for EtG of 274 ng/g and 547 cases with both FAEE- and EtG-negative, 33 cases with both FAEE- and EtG-positive, nine cases with FAEE-positive and EtG-negative, and seven cases with FAEE-negative and EtG-positive. Differences in physical, chemical, and biochemical properties and in the pharmacokinetic behavior are discussed as reasons for the deviating cases. In none of the 602 cases, serious alcohol consumption was reported by the mothers and no evidence for gestational ethanol exposure was observed in the medical investigation of the newborns. It is concluded that the combined use of FAEE and EtG in meconium as markers for fetal alcohol exposure essentially increases the accuracy of the interpretation and helps to avoid false positive and false-negative results.
脂肪酸乙酯（FAEE）和乙基葡萄糖醛酸苷（EtG）在一项母体健康评估研究中用于检测妊娠期间的酒精摄入，对 602 份胎粪样本进行了测定。采用经验证的顶空固相微萃取法与 GC-MS 联用，对 FAEE 进行了测定，并应用棕榈酸乙酯、亚油酸乙酯、油酸乙酯和硬脂酸乙酯的累积浓度（500ng/g 为截断值）进行解释。建立并验证了一种新的简单方法，用于从 10-20mg 胎粪中定量测定 EtG，以 D(5)-EtG 为内标，经甲醇/水（1:1，v/v）提取 30min，甲醇蒸发，纤维素滤器过滤水溶液，然后注入 LC-MS-MS。EtG 的检测限和定量限分别为 10 和 30ng/g，回收率为 86.6%至 106.4%，浓度的标准偏差范围为 37ng/g 时的 13%至 46,700ng/g 时的 5%（N=6）。在 43 例（7.1%）中发现 FAEE 超过截断值，累积浓度在 507 至 22,580ng/g 之间，有一例约 150,000ng/g（未检测到 EtG）。97 例（16.3%）检出 EtG，检出限至 10,200ng/g，另有一例 82,000ng/g（FAEE 为 10,500ng/g）。EtG 的截断值为 274ng/g 时，两种标志物之间的最佳一致性，547 例 FAEE 和 EtG 均为阴性，33 例 FAEE 和 EtG 均为阳性，9 例 FAEE 阳性而 EtG 阴性，7 例 FAEE 阴性而 EtG 阳性。讨论了物理、化学和生化性质以及药代动力学行为的差异，作为导致偏离病例的原因。在 602 例病例中，母亲均未报告严重饮酒，新生儿的医学检查也未观察到妊娠乙醇暴露的证据。结论是，FAEE 和 EtG 在胎粪中联合作为胎儿酒精暴露的标志物，可显著提高解释的准确性，并有助于避免假阳性和假阴性结果。
