Determination of ethyl glucuronide and ethyl sulfate as human biomarkers of alcohol exposure in urine by liquid chromatography/electrospray tandem mass spectrometry with large volume injection Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/fj236428q

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  • This research was undertaken to develop a sensitive, selective, and rapid analytical method to measure ethyl glucuronide (EtG) and ethyl sulfate (EtS) in urine as urinary biomarkers of alcohol exposure. Large-volume injection (LVI) coupled with liquid chromatography / electrospray tandem mass spectrometry was used to eliminate the need for either off-line or on-line solid phase extraction. Urine sample preparation was limited to diluting the urine 1:1000 with deionized water prior to large-volume injection (900 μL). Three MS/MS transitions were monitored for EtG and two MS/MS transitions were monitored for EtS, with the deprotonated molecular ion [M-H]- as the precursor ion: m/z 221→75, 221→85, 221→113(EtG) and 125→80, 125→97(EtS). Pentadeuterated EtG and pentadeuterated EtS were used as internal standards for EtG and EtS, respectively. The method was accurate with recoveries from 99.8% to 102.9% for EtG and 93.3% to 102.5% for EtS and gave good precision, as indicated by average RSD, of 3.5% and 4.9% for EtG and EtS, respectively. Matrix effects were eliminated and the instrumental detection limit was 0.005 μg/mL for EtG and 0.010 μg/mL for EtS, which is a factor of 10 lower than previous published analytical methods for measuring EtG and EtS. The method quantification limits were 0.015 μg/mL for EtG and 0.025 μg/mL for EtS in 1:1000 diluted urine, corresponding to 15 μg/mL for EtG and 25 μg/mL for EtS in undiluted urine. Finally, the developed method gave statistically equivalent results when applied to measuring EtG and EtS in a third-party reference urine.
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