Abstract
β-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, for detection of EtG none of the published LC-MS/MS methods could fulfill the minimum requirements of any of these guidelines. Therefore, an existing LC-MS/MS method has been modified by monitoring further MS/MS transitions instead of only one (deprotonated molecule [M − H]−/product ions: m/z 75, 85, 113, and 159 optional) with the aim of withstanding administrative or court scrutiny in forensic or workplace drug testing cases. Full method validation has been performed in accordance to guidelines of the German Society of Toxicology and Forensic Chemistry (GTFCh) and requirements of ISO 17025. One application field in the United States is a workplace monitoring program to detect surreptitious alcohol use among recovering health professionals, who by contract had agreed on total abstinence after drug and alcohol withdrawal therapy.
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Dahl, H.; Stephanson, N.; Beck, O.; Helander, A. Comparison of Urinary Excretion Characteristics of Ethanol and Ethylglucuronide. J. Anal. Toxicol. 2002,, 26, 201–204.
Kamil, I. A.; Smith, J. N.; Williams, R. T. A New Aspect of Ethanol Metabolism: Isolation of Ethylglucuronide. Biochem. J. 1952, 51, 32–33.
Janda, I.; Alt, A. Improvement of Ethylglucuronide Determination in Human Urine and Serum by Solid-Phase Extraction. J. Chromatogr. B Biomed. Sci. Appl. 2001, 758, 229–234.
Stephanson, N.; Dahl, H.; Helander, A.; Beck, O. Direct Quantification of Ethylglucuronide in Clinical Urine Samples by Liquid Chromatography-Mass Spectrometry. Therap. Drug Monitor. 2002, 24, 645–651.
Wurst, F. M.; Kempter, C.; Seidl, S.; Alt, A. Ethylglucuronide—A Marker of Alcohol Consumption and a Relapse Marker with Clinical and Forensic Implications. Alcohol Alcohol. 1999, 34, 71–77.
Wurst, F. M.; Schüttler, R.; Kempter, C.; Seidl, S.; Gilg, T.; Jachau, K.; Alt, A. Can Ethylglucuronide be Determined in Postmortem Body Fluids and Tissues?. Alcohol Alcohol. 1999, 34, 260–261.
Wurst, F. M.; Kempter, C.; Metzger, J.; Seidl, S.; Alt, A. Ethylglucuronide—A Marker of Recent Alcohol Consumption with Clinical and Forensic Implications. Alcohol 2000, 20, 111–116.
Wurst, F. M.; Metzger, J. W. On Behalf of the WHO/ISBRA Study on Biological State and Trait Markers of Alcohol Use and Dependence. The Direct Ethanol Metabolite Ethylglucuronide is a Specific Marker of Recent Alcohol Consumption. Alcohol Clin. Exp. Res. 2002, 26, 1114–1119.
Wurst, F. M.; Vogel, R.; Jachau, K.; Varga, A.; Alling, C.; Alt, A.; Skipper, G. E. Ethylglucuronide Discloses Recent Covert Alcohol Use Not Detected by Standard Testing in Forensic Psychiatric Inpatients. Alcohol Clin. Exp. Res. 2003, 27, 471–476.
Schmitt, G.; Droenner, P.; Skopp, G.; Aderjan, R. Ethylglucuronide: An Unusual Ethanol Metabolite in Humans. Synthesis, Analytical Data, and Determinaton in Serum and Urine. J. Anal. Toxicol. 1995, 19, 91–94.
Nishikawa, M.; Tsuchihashi, H.; Miki, A.; Katagi, M.; Schmitt, G.; Zimmer, H.; Keller, T.; Aderjan, R. Determination of Ethylglucuronide, a Minor Metabolite of Ethanol, in Human Serum by Liquid Chromatography-Electrospray Ionization Mass Spectrometry. J. Chromatogr. B Biomed. Sci. Appl. 1999, 726, 5–110.
Wurst, F. M.; Kempter, C.; Seidl, S.; Alt, A. Ethylglucuronide—A Marker of Alcohol Consumption and a Relapse marker with Clinical and Forensic Implications. Alcohol Alcohol. 1999, 34, 71–77.
Alt, A.; Wurst, F. M.; Seidl, S. Bestimmung von Ethylglucuronid in Urinproben mit dem internen Standard D5-Ethylglucuronid. Blutalkohol 1997, 34, 360–365.
Janda, I.; Weinmann, W.; Kuehnle, T.; Lahode, M.; Alt, A. Determination of Ethylglucuronide in Human Hair by SPE and LC-MS/MS. Forens. Sci. Int. 2002, 128, 59–65.
Alt, A.; Janda, I.; Seidl, S.; Wurst, F. M. Determination of Ethylglucuronide in Human Hair. Alcohol Alcohol. 2000, 35, 313–314.
Skopp, G.; Schmitt, G.; Pötsch, L.; Drönner, P.; Aderjan, R.; Mattern, R. Ethylglucuronide in Human Hair. Alcohol Alcohol. 2000, 35, 283–285.
Janda, I. Bestimmung des Alkoholkonsummarkers Ethylglucuronid in Körperflüssigkeiten, Haaren und Geweben—Methodenentwicklung und Validierung. Dissertation, University of Ulm, Germany, 2001; pp 85–88.
Skipper, G.E.; Schaefer, P.; Thierauf, A.; Weinmann, W.; Dierkes, J.; Wurst, F.M. Detection of Surreptitious Alcohol Use Among Health Professionals Recovering from Substance-Related Disorders Using a New Marker, Ethyl Glucuronide, unpublished
Schmitt, G.; Aderjan, R.; Kirchner, M.; Zimmer, H. Enzymimmunoassay und GC-MS zur Bestimmung von Ethylglucuronid in Serum und Urin. Blutalkohol 1998, 25, 428–433.
Zimmer, H.; Schmitt, G.; Aderjan, R. Preliminary Immunochemical Test for the Determination of Ethylglucuronide in Serum and Urine: Comparison of Screening Method Results with Gas Chromatography-Mass Spectrometry. J. Anal. Toxicol. 2002, 26, 11–16.
Wurst, F. M.; Metzger, J.; Jauchau, K.; Seidl, S.; Pridzun, L.; Janda, I.; Alt, A. The Direct Ethanol Metabolite Ethylglucuronide: A Specific Marker of Recent Alcohol Consumption. In New and Upcoming Markers of Alcohol Consumption; Wurst, F. M., Ed.; Steinkopff-Springer Verlag: Darmstadt, 2001; 62–74.
Shoemaker, M. J.; Earley, R. J.; Gorsky, J. E.; Jenny, R. W.; Leiendecker-Foster, C.; Markus, W. R.; Moore, N.; Welch, M. J. Urine Drug Testing in the Clinical Laboratory; Approved Guideline, 1999. (http://www.nccls.org/free/TDM8-A.pdf accessed May 20, 2003)
Aderjan, R.; Babel, B.; Briellmann, T.; Daldrup, T.; Demme, U.; Hallbach, J.; Hartung, M.; Harzer, K.; Herbold, M.; von Meyer, L.; Moeller, M.; Musshoff, F.; Schmitt, G.; Weinmann, W. Anlage zu den Richtlinien der GTFCh zur Qualitätssicherung bei forensisch-toxikologischen Untersuchungen. Anhang A: Anforderung an einzelne Analysenmethoden. Toxichem + Krimtech 2000, 67, 13–16. (http://www.gtfch.org/GTFChneuEngl.htm accessed May 20, 2003).
Stolker, A. A. M.; Stephany, R. W.; van Ginkel, L. A. Identification of Residues by LC-MS. The Application of New EU Guidelines. Analusis 2000, 28, 947–951.
Rivier, L. Criteria for the Identification of Compounds by Liquid Chromatography-Mass Spectrometry and Liquid Chromatography-Multiple Mass Spectrometry in Forensic Toxicology and Doping Analysis. Anal. Chimica Acta 2002, 492, 69–82.
Schmidt, G.; Herbold, M.; Peters, F. Qualitaetssicherung im forensich-toxikologichen Labor and VALISTAT 1.0. (http://www.pts-gtfch.de/topd.htm).
Weinmann, W.; Svoboda, M. Fast Screening for Drugs of Abuse by Solid-Phase Extraction Combined with Flow-Injection Ionspray Tandem Mass Spectrometry. J. Anal. Toxicol. 1998, 22, 319–328.
Weinmann, W.; Vogt, S.; Goerke, R.; Mueller, C.; Bromberger, A. Simultaneous Determination of THC-COOH and THC-COOH-Glucuronide in Urine Samples by LC-MS/MS. Forens. Sci. Int. 2000, 113, 381–387.
Peters, F. T.; Maurer, H. H. Bioanalytical Method Validation and Its Implications for Forensic and Clinical Toxicology—A Review. Accred. Qual. Assur. 2002, 7, 441–449.
Dahl, H.; Stephanson, N.; Beck, O.; Helander, A. Comparison of Urinary Excretion Characteristics of Ethanol and Ethyl Glucuronide. J. Anal. Toxicol. 2002, 26, 201–204.
Seidl, S.; Wurst, F. M.; Alt, A. Überprüfung einer Abstinenzbehauptung in der Fahreignungsoberbegutachtung mit Hilfe des Ethanolmetaboliten Ethylglucuronid (EtG). Blutalkohol 1998, 35, 174–182.
Sticht, G. GC-MS-Bestimmung von Ethylglucuronid im Serum mit Propylglucuronid als internem Standard. Rechtsmedizin 1997, 7, 184–188.
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Published online December 17, 2003.
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Weinmann, W., Schaefer, P., Thierauf, A. et al. Confirmatory analysis of ethylglucuronide in urine by liquid-chromatography/electrospray ionization/tandem mass spectrometry according to forensic guidelines. J Am Soc Mass Spectrom 15, 188–193 (2004). https://doi.org/10.1016/j.jasms.2003.10.010
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DOI: https://doi.org/10.1016/j.jasms.2003.10.010