Toxicological Centre, University of Antwerp, Antwerp, Belgium.
Forensic Sci Int. 2010 Mar 20;196(1-3):121-7. doi: 10.1016/j.forsciint.2009.12.043. Epub 2010 Jan 12.
Ethyl glucuronide (EtG) detection in non-conventional matrices, such as hair and meconium, can provide useful information on alcohol abuse over a long time frame, for example during pregnancy or after a withdrawal treatment. This study reports on the development, validation and application of a new hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for the analysis of EtG in meconium and hair. For each matrix, the sample preparation and the chromatographic separation were thoroughly optimised. Additionally, experiments with reversed-phase liquid chromatography were also performed in the development stages. Analyses were carried out using a Phenomenex Luna HILIC column (150 mm x 3 mm, 5 microm) and a mobile phase composed by ammonium acetate 2mM and acetonitrile, in gradient. Different SPE cartridges (Oasis MAX, Oasis WAX, aminopropyl silica) and solvents were tested in order to obtain the highest recoveries and cleanest extracts. Optimal results were obtained for meconium with aminopropyl cartridges, while for hair an incubation of 16 h with 2 mL of water and acetonitrile (50/50, v/v) provided good results. The analytical method was validated for both matrices (meconium and hair) by assessing linearity, precision, accuracy, recovery and limit of quantification. The calibration curve concentrations ranged from 50 to 1200 pg/mg for meconium and from 20 to 1000 pg/mg for hair. Real meconium and hair samples were analyzed and results were consistent with literature.
乙基葡糖苷(EtG)在非传统基质(如头发和胎粪)中的检测可提供关于长时间内滥用酒精的有用信息,例如在怀孕期间或戒酒后。本研究报告了一种新的亲水相互作用液相色谱-串联质谱(HILIC-MS/MS)方法的开发、验证和应用,用于分析胎粪和头发中的 EtG。对于每种基质,都对样品制备和色谱分离进行了彻底优化。此外,在开发阶段还进行了反相液相色谱实验。分析使用 Phenomenex Luna HILIC 柱(150mmx3mm,5μm)和由 2mM 乙酸铵和乙腈组成的流动相在梯度条件下进行。为了获得最高的回收率和最干净的提取物,测试了不同的 SPE 小柱(Oasis MAX、Oasis WAX、氨基丙基硅胶)和溶剂。对于胎粪,氨基丙基小柱获得了最佳结果,而对于头发,用 2mL 水和乙腈(50/50,v/v)孵育 16 小时可获得良好结果。通过评估线性、精密度、准确度、回收率和定量限,对两种基质(胎粪和头发)的分析方法进行了验证。胎粪的校准曲线浓度范围为 50-1200pg/mg,头发的浓度范围为 20-1000pg/mg。分析了真实的胎粪和头发样本,结果与文献一致。