Development of LC-MS/MS Method of Measurement of Direct Ethanol Biomarkers Ethyl Glucuronid and Ethyl Sulfate in Urine and Serum Matriches in Detection and Tracking of Alcohol Use
PDF
Cite
Share
Request
Research Article
P: 122-127
June 2023

Development of LC-MS/MS Method of Measurement of Direct Ethanol Biomarkers Ethyl Glucuronid and Ethyl Sulfate in Urine and Serum Matriches in Detection and Tracking of Alcohol Use

J Ankara Univ Fac Med 2023;76(2):122-127
1. Acıbadem Üniversitesi Tıp Fakültesi, Adli Tıp Anabilim Dalı, İstanbul, Türkiye
2. SEM Laboratuvar Cihazları Pazarlama San. ve Tic. A.Ş. Ar-Ge Merkezi, İstanbul, Türkiye
No information available.
No information available
Received Date: 21.04.2022
Accepted Date: 13.04.2023
Publish Date: 31.07.2023
PDF
Cite
Share
Request

ABSTRACT

Objectives:

Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are minor non-oxidative metabolites of ethanol. They can be detected in body fluids shortly after ethanol consumption and have a longer detection window than the parent compound. They are considered highly sensitive and specific biomarkers for recent alcohol consumption. EtG and EtS biomarkers are often used in the clinical and justice system to monitor alcohol withdrawal. In this study, it was aimed to develop an easy and cost-effective analysis methodology that provides the qualitative and quantitative detection of EtG and EtS in urine and serum matrices.

Materials and Methods:

For the simultaneous quantification of EtG and EtS biomarkers in urine and serum, an LC-MS/MS method with fast and reliable sample preparation protocols with 5 minutes analysis time has been developed and validated. Two product ions along with the precursor ions of both alcohol metabolites were monitored and the deuteron-derivative of EtG was used as an internal standard, for increasing the reliability of the results.

Results:

The validation parameters and obtained values were investigated based on international guidelines.They were found within acceptable ranges. Thus, sensitive determination of EtG and EtS in urine/serum samples was achieved with a simple sample preparation protocol which could be automated.

Conclusion:

In this study, a simple LC-MS/MS based analytical methodology has been developed for the simultaneous quantitative analysis of EtG and EtS in urine and serum matrices. Time-consuming and costly processes such as chemical modification (derivatization), matrix cleaning with SPE cartridges or concentration were not applied at any step of sample preparation. The developed analytical method has been validated within the framework of internationally accepted biochemical analytical method development guidelines.

Keywords: Alcohol Biomarkers, Ethyl Glucuronide, Ethyl Sulphate, Alcohol Use Disorder, LC-ESI-MS/MS

References

1
Halter CC, Dresen S, Auwaerter V, Wurst FM, Weinmann W. Kinetics in serum and urinary excretion of ethyl sulfate and ethyl glucuronide after medium dose ethanol intake. Int J Legal Med. 2008;122:123-128.
2
Luginbühl M, Weinmann W, Butzke I, Pfeifer P. Monitoring of direct alcohol markers in alcohol use disorder patients during withdrawal treatment and successive rehabilitation. Drug Test Anal. 2019;11:859-869.
3
Barrio P, Teixidor L, Rico N, et al. Urine Ethyl Glucuronide Unraveling the Reality of Abstinence Monitoring in a Routine Outpatient Setting: A Cross-Sectional Comparison with Ethanol, Self Report and Clinical Judgment. Eur Addict Res. 2016;22:243-248.
4
Reisfield GM, Teitelbaum SA, Opie SO, Jones J, Morrison DG, Lewis B. The roles of phosphatidylethanol, ethyl glucuronide, and ethyl sulfate in identifying alcohol consumption among participants in professionals health programs. Drug Test Anal. 2020;12:1102-1108.
5
Ulwelling W, Smith K. The PEth Blood Test in the Security Environment: What it is; Why it is Important; and Interpretative Guidelines. J Forensic Sci. 2018;63:1634-1640.
6
Fucci N, Gili A, Aroni K, et al. Monitoring people at risk of drinking by a rapid urinary ethyl glucuronide test. Interdiscip Toxicol. 2017;10:155-162.
7
Walsham NE, Sherwood RA. Ethyl glucuronide. Ann Clin Biochem. 2012;49:110-117.
8
Grodin EN, Nguyen XT, Ho D, Bujarski S, Ray LA. Sensitivity and specificity of a commercial urinary ethyl glucuronide (ETG) test in heavy drinkers. Addict Behav Rep. 2020;11:100249.
9
Hernández Redondo A, Schroeck A, Kneubuehl B, Weinmann W. Determination of ethyl glucuronide and ethyl sulfate from dried blood spots. Int J Legal Med. 2013;127:769-775.
10
Aşıcıoğlu F, Mutlu E, Belce A. The Importance of Ethyl Glucuronide and Ethyl Sulphate Analysis in Forensic Cases. Turkiye Klinikleri J Foren Sci Leg Med. 2019;16:64-69.
11
Helander A, Olsson I, Dahl H. Postcollection synthesis of ethyl glucuronide by bacteria in urine may cause false identification of alcohol consumption. Clin Chem. 2007;53:1855-1857.
12
Arndt T, Beyreiß R, Schröfel S, Stemmerich K. Cross-reaction of propyl and butyl alcohol glucuronides with an ethyl glucuronide enzyme immunoassay. Forensic Sci Int. 2014;241:84-86.
13
Arndt T, Grüner J, Schröfel S, Stemmerich K. False-positive ethyl glucuronide immunoassay screening caused by a propyl alcohol-based hand sanitizer. Forensic Sci Int. 2012;223:359-363.
14
Arndt T, Gierten B, Güssregen B, Werle A, Grüner J. False-positive ethyl glucuronide immunoassay screening associated with chloral hydrate medication as confirmed by LC-MS/MS and self-medication. Forensic Sci Int. 2009;184:e27-29.
15
Berg T, Eliassen E, Jørgenrud B, Kabashi S, Petukhov A, Bogstrand ST. Determination of phosphatidylethanol 16:0/18:1 in whole blood by 96-well supported liquid extraction and UHPLC-MS/MS. J Clin Lab Anal. 2019;33:e22631.
2024 ©️ Galenos Publishing House