Determination of ethyl glucuronide, a minor metabolite of ethanol, in human serum by liquid chromatography–electrospray ionization mass spectrometry

Nishikawa, Mayumi; Tsuchihashi, Hitoshi; Miki, Akihiro; Katagi, Munehiro; Schmitt, Georg; Zimmer, Holger; Keller, Th.; Aderjan, R.

doi:10.1016/s0378-4347(99)00008-0

Cited by 64 publications

(43 citation statements)

References 9 publications

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“…After 5.5 h, EtG could be monitored in the sera of volunteers A and B, but not for C, who ingested the lowest amount of ethanol. These data compare well with those found in other studies [5,14]. Furthermore, with one exception (lactate of volunteer C), the concentrations of lactate and acetate became enhanced upon alcohol consumption.…”

Section: Kinetics Of Etg Lactate and Acetate After Ethanol Ingestionsupporting

confidence: 91%

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Effects of lactate and acetate on the determination of serum ethyl glucuronide by CZE

et al. 2006

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The analysis of ethyl glucuronide (EtG), a marker of recent alcohol consumption, in serum with an optimized CZE assay is reported. The method uses a 0.1-mm id fused-silica capillary of 50 cm effective length that is coated with linear polyacrylamide, a pH 4.4 nicotinic acid/epsilon-aminocaproic acid (EACA) BGE, reversed polarity and indirect analyte detection. The assay is based on a 1:1 dilution of serum with deionized water and has LODs for EtG, lactate and acetate of 3.8 x 10(-7) M, 2.60 x 10(-6 )M and 2.18 x 10(-6 )M, respectively. Separation of EtG from endogenous macro- and microcomponents (anionic serum components of high and low concentration, respectively) and its quantification are shown to be possible for a wide range of lactate (stacker) and acetate (destacker) concentrations, macrocomponents that have an impact on the CZE behavior of EtG and that change after intake of ethanol. The assay has been successfully applied to the analysis of EtG, lactate and acetate in (i) sera of volunteers that ingested known amounts of alcohol and (ii) samples of patients that were classified (teetotalers and social drinkers vs. alcohol abusers) via analysis of carbohydrate-deficient transferrin.

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Section: Kinetics Of Etg Lactate and Acetate After Ethanol Ingestionsupporting

confidence: 91%

“…For analysis of urine, human serum, hair and tissues (liver, brain, fat), GC-MS after derivatization, and LC-MS or LC-MS/MS with negative ESI were used [14][15][16][17]. Recently, a simple, fast and inexpensive CZE procedure enabling the determination of EtG in 1:1 diluted serum with an LOD of 3.56610 27 M was reported [18].…”

Section: Introductionmentioning

confidence: 99%

Effects of lactate and acetate on the determination of serum ethyl glucuronide by CZE

et al. 2006

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“…In urine, it can be detected longer than ethanol. Therefore, EtG meets the need for a sensitive and specific marker to elucidate alcohol use not detected by standard testing [5][6][7][8][9][10][11][12][13]. Furthermore, EtG has been detected in hair samples [14 -16] and post-mortem tissues (liver, brain, fat tissue [17]) of alcohol addicts.…”

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confidence: 99%

“…For the identification of residues by LC-MS/MS analysis, European guidelines require chromatographic separation and a minimum of two transitions for substance detection; when LC-MS (single MS) is used, three characteristic ions are required [24,25]. None of the published LC-MS or LC-MS/MS methods for EtG listed in Table 1 fulfilled these requirements, since either only one ion with optimized "in-source" CID conditions was used with a single-quadrupole instrument [4,11], or only one …”

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confidence: 99%

Confirmatory analysis of ethylglucuronide in urine by liquid-chromatography/electrospray ionization/tandem mass spectrometry according to forensic guidelines

Weinmann¹,

Schaefer²,

Thierauf³

et al. 2004

J. Am. Soc. Mass Spectrom.

125

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␤-D-ethylglucuronide (EtG) is a stable Phase II metabolite of ethanol which can be detected in urine samples several days after elimination of ethanol. It is a useful diagnostic parameter for monitoring abstinence of alcoholics in alcohol withdrawal treatment. For this purpose, determination in urine is mainly performed by LC-MS, LC-MS/MS, or by GC-MS. For the mass spectrometric identification and detection of controlled substances in more sensitive fields such as forensic toxicology, workplace drug testing, doping analysis, and veterinary organic residue control, official guidelines have been released requiring a chromatographic separation and a minimum of two mass spectrometric transitions of the analyte. However alcohol consumption remain suboptimal with regard to sensitivity and specificity. Furthermore, these biomarkers can be influenced by age, gender, and a variety of substances and non-alcohol-associated diseases, and do not fully cover the time axis for alcohol intake. Conjugation of ethanol with activated glucuronic acid in the presence of membrane-bound mitochondrial UDP glucuronyl transferase represents a minor detoxification pathway for ethanol: About 0.02-0.06% (mean) of the dose of ethanol administered is recovered as ␤-D-ethylglucuronide (EtG) in urine in humans [1] and-dose dependent-0.5-1.5% in rabbits [2]. EtG is a non-volatile, water-soluble, stable, direct metabolite of ethanol that can be detected in various body fluids, tissues and hair. EtG (C 8 H 14 O 7 ) has a molecular weight of 222 g/mol, and the melting point (decomposition temperature) is about 150°C. Shortly after the initial consumption of even small amounts of ethanol, EtG is formed. It has been detected in urine up to 80 h after the complete elimination of alcohol from the body and was not detectable in teetotalers with a 0.1 mg/L cut-off [3,4]. EtG is unique in covering this important time span of one to three days after alcohol uptake. In urine, it can be detected longer than ethanol. Therefore, EtG meets the need for a sensitive and specific marker to elucidate alcohol use not detected by

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“…Nishikawa et al carried out EtG analysis in human serum by liquid chromatographyelectrospray ionization mass spectrometry. 13 However, this method is typically used to analyze a single target material or chemical and requires sample processing prior to analysis. Current mass spectroscopy methods are also laborious and expensive.…”

Section: Introductionmentioning

confidence: 99%

1D Proton NMR Spectroscopic Determination of Ethanol and Ethyl Glucuronide in Human Urine

Kim¹,

Lee²,

Yoon³

et al. 2013

Bulletin of the Korean Chemical Society

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Forensic and legal medicine require reliable data to indicate excessive alcohol consumption. Ethanol is oxidatively metabolized to acetate by alcohol dehydrogenase and non-oxidatively metabolized to ethyl glucuronide (EtG), ethyl sulfate (EtS), phosphatidylethanol, or fatty acid ethyl esters (FAEE). Oxidative metabolism is too rapid to provide biomarkers for the detection of ethanol ingestion. However, the nonoxidative metabolite EtG is a useful biomarker because it is stable, non-volatile, water soluble, highly sensitive, and is detected in body fluid, hair, and tissues. EtG analysis methods such as mass spectroscopy, chromatography, or enzyme-linked immunosorbent assay techniques are currently in use. We suggest that nuclear magnetic resonance (NMR) spectroscopy could be used to monitor ethanol intake. As with current conventional methods, NMR spectroscopy doesn't require complicated pretreatments or sample separation. This method has the advantages of short acquisition time, simple sample preparation, reproducibility, and accuracy. In addition, all proton-containing compounds can be detected. In this study, we performed 1 H NMR analyses of urine to monitor the ethanol and EtG. Urinary samples were collected over time from 5 male volunteers. We confirmed that ethanol and EtG signals could be detected with NMR spectroscopy. Ethanol signals increased immediately upon alcohol intake, but decreased sharply over time. In contrast, EtG signal increased and reached a maximum about 9 h later, after which the EtG signal decreased gradually and remained detectable after 20-25 h. Based on these results, we suggest that 1 H NMR spectroscopy may be used to identify ethanol non-oxidative metabolites without the need for sample pretreatment.

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Determination of ethyl glucuronide, a minor metabolite of ethanol, in human serum by liquid chromatography–electrospray ionization mass spectrometry

Cited by 64 publications

References 9 publications

Effects of lactate and acetate on the determination of serum ethyl glucuronide by CZE

Effects of lactate and acetate on the determination of serum ethyl glucuronide by CZE

Confirmatory analysis of ethylglucuronide in urine by liquid-chromatography/electrospray ionization/tandem mass spectrometry according to forensic guidelines

1D Proton NMR Spectroscopic Determination of Ethanol and Ethyl Glucuronide in Human Urine

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