Validated Method for the Determination of the Ethanol Consumption Markers Ethyl Glucuronide, Ethyl Phosphate, and Ethyl Sulfate in Human Urine by Reversed-Phase/Weak Anion Exchange Liquid Chromatography−Tandem Mass Spectrometry

A liquid chromatography−tandem mass spectrometry (LC−MS/MS) method for the determination of the ethanol consumption markers ethyl glucuronide (EtG), ethyl phosphate (EtP), and ethyl sulfate (EtS) in human urine was developed. A reversed-phase/weak anion exchange type stationary phase demonstrated particular suitability for the analysis of these highly polar acids. Sample preparation was minimized to centrifugation and dilution of urine prior to injection. The method was validated in the range of 5−750 μg·L-1 with 1:20 and 1:1000 diluted urine, which corresponds to actual concentration ranges of 0.1−15 and 5−750 mg·L-1 in undiluted samples. Method validation was carried out using six different lots of human urine. Over the entire calibration range intraday and interday precision (each n = 5, three concentration levels per dilution factor) adopted values between 0.6 and 4.7% and 0.8 and 12.1%, relative standard deviation, respectively. Corresponding accuracy values ranged between 94.2 and 113.5% and 86.6 and 110.9%, respectively. Matrix effects (absolute/relative) were found to be present in minor extent (∼ −30% to +15% MS/MS signal alterations) and were well corrected by the employed isotopically labeled internal standards. The validated assay was applied to urine samples of a drinking study as well as postmortem specimens. It was possible to assess the principal potential of EtP as ethanol consumption marker. Elevated concentration levels were found in real samples; however, EtP seems to be less sensitive compared to the previously known ethanol phase II conjugates EtG and EtS.

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PID https://www.doi.org/10.1021/ac060680+.s001
URL https://figshare.com/articles/journal_contribution/Validated_Method_for_the_Determination_of_the_Ethanol_Consumption_Markers_Ethyl_Glucuronide_Ethyl_Phosphate_and_Ethyl_Sulfate_in_Human_Urine_by_Reversed_Phase_Weak_Anion_Exchange_Liquid_Chromatography_Tandem_Mass_Spectrometry/3371803
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Author Bicker, Wolfgang
Author Lämmerhofer, Michael
Author Keller, Thomas
Author Schuhmacher, Rainer
Author Krska, Rudolf
Author Lindner, Wolfgang
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Collected From figshare
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Publication Date 2016-05-12
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Language English
Resource Type Other literature type
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Source https://science-innovation-policy.openaire.eu/search/publication?articleId=r37980778c78::ccabfe812f9f9ddb31c9b393f1e525bf
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Last Updated 25 December 2020, 17:53 (CET)
Created 25 December 2020, 17:53 (CET)