Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Artillerigatan 12, SE-587 58 Linkoping, Sweden.
Forensic Sci Int. 2012 Feb 10;215(1-3):51-5. doi: 10.1016/j.forsciint.2011.01.044. Epub 2011 Mar 1.
The overall objectives of the study were to develop a sensitive method for ethyl glucuronide (EtG) determination in hair and then investigate if a low or moderate intake of ethanol could be differentiated from total abstinence. Forty-four subjects were included in the study, 12 males (7 drinkers and 5 abstinent) and 32 females (14 drinkers and 18 abstinent). The study lasted 3 months and the female drinkers consumed one glass (16 g of ethanol) and the males consumed two glasses (32 g of ethanol) of wine (13.5-14%) daily. Hair samples were collected as close as possible above the skin and the proximal 2 cm were analyzed for EtG. Hair was cut into pieces of about 0.5 cm length and washed before incubation overnight in water and then extracted on Clean Screen EtG Carbon columns. The LC/MS/MS system consisted of a Waters ACQUITY UPLC connected to an API 4000 triple quadrupole instrument. Two transitions for EtG and one for the internal standard EtG-D(5) were measured. The method was linear from 60 to 10,000 pg/sample. Imprecision studies were performed at three levels as well as with an authentic sample. Total imprecision was 16% at 200 pg/sample, 8% at 1000 pg/sample, 6% at 8000 pg/sample and 13% at 29 pg/mg in the authentic sample. Of those who drank two glasses of wine every day, four had measurable amounts of EtG in their hair (5-11 pg/mg), and in only one of the females drinking one glass of wine EtG was quantified (3 pg/mg). Among the 23 abstinent subjects two had traces of EtG in the hair. We conclude that persons who ingested 16 or 32 g of ethanol daily for 3 months presented with low concentrations of EtG in hair, well below the proposed threshold for overconsumption set at 30 pg/mg. In addition, none of those who ingested 16 g/day had concentrations over the proposed abstinence threshold of 7 pg/mg.
本研究的总体目标是开发一种灵敏的毛发乙基葡萄糖醛酸苷（EtG）检测方法，然后研究能否区分低或中量乙醇摄入与完全戒断。研究纳入 44 名受试者，12 名男性（7 名饮酒者和 5 名戒酒者）和 32 名女性（14 名饮酒者和 18 名戒酒者）。研究持续 3 个月，女性饮酒者每天饮用一杯（16 克乙醇），男性饮酒者每天饮用两杯（32 克乙醇）葡萄酒（13.5-14%）。尽可能靠近头皮采集毛发样本，分析近端 2 厘米处的 EtG。毛发剪成约 0.5 厘米长的小段，在水中孵育过夜，然后用 Clean Screen EtG Carbon 柱提取。LC/MS/MS 系统由 Waters ACQUITY UPLC 与 API 4000 三重四极杆仪器相连。测量 EtG 和内标 EtG-D(5)的两个跃迁。方法在 60 至 10000 pg/样品的范围内呈线性。在三个水平和一个真实样本中进行了精密度研究。在 200 pg/样品时总精密度为 16%，在 1000 pg/样品时为 8%，在 8000 pg/样品时为 6%，在真实样本中为 29 pg/mg 时为 13%。在每天饮用两杯葡萄酒的人群中，有 4 人的头发中可测量到 EtG（5-11 pg/mg），而在每天饮用一杯葡萄酒的女性中，只有 1 人定量检测到 EtG（3 pg/mg）。在 23 名戒酒者中，有 2 人的头发中存在 EtG 痕迹。我们的结论是，每天摄入 16 或 32 克乙醇持续 3 个月的人，其头发中的 EtG 浓度较低，远低于设定的 30 pg/mg 过量摄入阈值。此外，每天摄入 16 克乙醇的人，其浓度均未超过设定的 7 pg/mg 戒酒阈值。
