Kinetics in serum and urinary excretion of ethyl sulfate and ethyl glucuronide after medium dose ethanol intake

Institute of Forensic Medicine, University Hospital Freiburg, Albertstrasse 9, 79104 Freiburg, Germany.
Deutsche Zeitschrift für die Gesamte Gerichtliche Medizin (Impact Factor: 2.71). 03/2008; 122(2):123-8. DOI: 10.1007/s00414-007-0180-8
Source: PubMed


The direct ethanol metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS), are of increasing importance for clinical and forensic applications, but there are only few studies on the kinetics of EtG in serum and none on EtS. In this study, 13 volunteers (social drinkers) drank ethanol in the form of white wine to reach a blood alcohol concentration of 0.51 +/- 0.17 g/kg, and blood and urine samples were analyzed for EtG and EtS simultaneously by chromatography-tandem mass spectrometry (LC-MS/MS). Mean peak serum EtG and EtS concentrations were 2.9 +/- 1.3 and 2.8 +/- 1.6 micromol/l, respectively, and were reached between 4.0 +/- 0.9 h after the start of drinking (3.0 +/- 0.5 h for EtS). The mean time differences between reaching maximum blood ethanol levels and serum metabolite levels were 2.3 +/- 0.9 h for EtG and 1.2 +/- 0.5 h for EtS. In the last blood samples collected (10-11 h after the start of drinking), 11 (of 13) volunteers were still positive for EtG in serum, whereas only 2 were positive for EtS. In the serum of one female person, no EtS was detectable at any time; however, it was excreted in the urine in (low) concentrations. Ethanol was detectable in the serum for up to 8.6 h after the start of drinking, whereas EtG and EtS were detectable up to more than 5.8 h (EtG) and 4.0 h (EtS), respectively. Mean peak urinary concentrations were 401 +/- 232 micromol/l for EtG and 266 +/- 153 micromol/l for EtS, and mean peak levels were reached 6.2 +/- 0.9 h (EtG) and 5.3 +/- 1.2 h (EtS) after the start of drinking. Maximum concentrations of EtG and EtS in serum showed a wide interindividual variation and could not be correlated to the maximum blood ethanol concentrations. Correlations (p < 0.001, Kendall's Tau b) were found when comparing pairs of parameters, but mostly involved areas under the curve (AUC) of metabolites or of ethanol; one correlation linked the peak concentrations of EtG and EtS in urine.

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    • "Although combined tests, like early detection of alcohol consumption (EDAC) and an analysis of a panel of 10–56 routine laboratory tests, are considered useful in detection of heavy drinking (Harasymiw and Bean, 2001, 2007; Helander, 2003; Harasymiw et al., 2004, 2005; Bean and Harasymiw, 2011), more specific tests seem to be much more convenient. Determination of the ethanol derivatives of extended excretion time, like ethyl glucuronides and sulfates, fatty acid ethyl esters and phosphatidylethanol , is considered as they are sensitive and specific indicators of recent alcohol ingestion (Dahl et al., 2002; Helander and Beck, 2004, 2005; Erim et al., 2007; Halter et al., 2008; Helander and Zheng, 2009; Helander et al., 2009a,b; Palmer, 2009; Waszkiewicz et al., 2010b; Hastedt et al., 2012; Lees et al., 2012). Although useful, such tests also generate false-positive as well as false-negative results. "
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    ABSTRACT: Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. In lectin blotting ten glycoprotein bands were analyzed. The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake.
    Alcohol and Alcoholism 10/2013; 49(1). DOI:10.1093/alcalc/agt152 · 2.89 Impact Factor
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    • "Some reports have indeed documented a high interindividual variability in EtG production (Halter et al., 2008; Paul et al., 2008). Halter et al. (2007) showed that the administration of a conventional dose of ethanol to 13 individuals resulted in highly variable (8-fold) maximum concentrations of serum EtG, and that EtG concentrations did not correlate with blood ethanol concentrations (Halter et al., 2008). This marked interindividual variability in EtG levels could be attributed to variable activities of UGTs involved in ethanol metabolism. "
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    ABSTRACT: Background: Ethylglucuronide (EtG) determination is increasingly used in clinical and forensic toxicology to document ethanol consumption. The enzymes involved in EtG production, as well as potential interactions with common drugs of abuse, have not been extensively studied. Methods: Activities of human liver (HLM), kidney (HKM) and intestinal (HIM) microsomes, as well as of twelve major human recombinant UDP-glucuronosyltransferases (UGTs), toward ethanol (50 and 500 mM) were evaluated in vitro using liquid chromatography-tandem mass spectrometry. Enzyme kinetic parameters were determined for pooled microsomes and recombinant UGTs with significant activity. Individual contributions of UGTs were estimated using the relative activity factor (RAF) approach, proposed for scaling activities obtained with cDNA-expressed enzymes to HLM. Interaction of morphine, codeine, lorazepam, oxazepam, nicotine, cotinine, cannabinol and cannabidiol (5, 10, 15 mg/L) with ethanol (1.15, 4.6, 11.5 g/L; i.e. 25, 100, 250 mM) glucuronidation was assessed using pooled HLM. Results: Ethanol glucuronidation intrinsic clearance (Cl(int)) was 4- and 12.7- times higher for HLM than for HKM and HIM, respectively. All recombinant UGTs, except UGT1A1, 1A6 and 1A10, produced EtG in detectable amounts. UGT1A9 and 2B7 were the most active enzymes, each accounting for 17% and 33% of HLM Cl(int), respectively. Only cannabinol and cannabidiol significantly affected ethanol glucuronidation. Cannabinol increased ethanol glucuronidation in a concentration-dependent manner, whereas cannabidiol significantly inhibited EtG formation in a non-competitive manner (IC(50)=1.17 mg/L; Ki=3.1 mg/L). Conclusions: UGT1A9 and 2B7 are the main enzymes involved in ethanol glucuronidation. In addition, our results suggest that cannabinol and cannabidiol could alter significantly ethanol glucuronidation.
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    • "According to former investigations [24] [29] [30], the marker and ethanol concentrations did not show good analogy and vice versa. "
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    Forensic science international 02/2011; 210(1-3):63-8. DOI:10.1016/j.forsciint.2011.01.036 · 2.14 Impact Factor
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