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用于揭示近期酒精摄入量的生物标志物:采用新型直接超高效液相色谱-串联质谱法和酶联免疫吸附测定法检测5-羟色醇葡糖醛酸苷的潜力

Biomarkers to disclose recent intake of alcohol: potential of 5-hydroxytryptophol glucuronide testing using new direct UPLC-tandem MS and ELISA methods.

作者信息

Beck Olof, Stephanson Nikolai, Böttcher Michael, Dahmen Norbert, Fehr Christoph, Helander Anders

机构信息

Department of Medicine, Karolinska Institute and Karolinska University Hospital, Stockholm, Sweden.

出版信息

Alcohol Alcohol. 2007 Jul-Aug;42(4):321-5. doi: 10.1093/alcalc/agm039. Epub 2007 May 28.

Abstract

AIMS

This study compared two new methods for direct determination of 5-hydroxytryptophol glucuronide (GTOL) in urine, a biomarker for detection of recent alcohol consumption.

METHODS

Urine samples were collected from ten alcoholic patients during recovery from intoxication. A direct injection ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for measurement of the urinary GTOL to 5-hydroxyindoleacetic acid (5-HIAA) ratio, and an ELISA assay for direct measurement of GTOL, were used. Comparison was made with the urinary ethanol and ethyl glucuronide (EtG) concentrations.

RESULTS

The breath ethanol concentration on admission ranged between 1.0-3.1 g/l. The UPLC-MS/MS method showed a median detection time of 39 h for an elevated urinary GTOL/5-HIAA ratio, while EtG was detected for a median of 65 h. Determination of GTOL by the ELISA assay showed 87% sensitivity in detecting positive samples at a 44% specificity, as compared with the UPLC-MS/MS method.

CONCLUSIONS

The lower sensitivity of the urinary GTOL/5-HIAA ratio compared with EtG for recent drinking may be clinically useful, in cases where the EtG test provides an unwanted high sensitivity for intake of only small amounts of alcohol or unintentional ethanol exposure.

摘要

目的

本研究比较了两种直接测定尿中5-羟色胺醇葡糖苷酸(GTOL)的新方法,GTOL是近期饮酒检测的生物标志物。

方法

从10名酒精中毒恢复期的患者收集尿液样本。采用直接进样超高效液相色谱-串联质谱法(UPLC-MS/MS)测定尿中GTOL与5-羟吲哚乙酸(5-HIAA)的比值,以及采用酶联免疫吸附测定法(ELISA)直接测定GTOL。并与尿中乙醇和葡糖醛酸乙酯(EtG)浓度进行比较。

结果

入院时呼气乙醇浓度在1.0 - 3.1 g/l之间。UPLC-MS/MS方法显示,尿中GTOL/5-HIAA比值升高的中位检测时间为39小时,而EtG的中位检测时间为65小时。与UPLC-MS/MS方法相比,ELISA法测定GTOL在检测阳性样本时的灵敏度为87%,特异性为44%。

结论

与EtG相比,尿中GTOL/5-HIAA比值对近期饮酒的较低灵敏度在临床上可能有用,例如在EtG检测对仅少量饮酒或无意乙醇暴露摄入具有过高灵敏度的情况下。

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