Research Article

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


  • Hazal Yılmaz
  • Murat Emrah Maviş
  • Gökçe Göksu Gürsu
  • Oğuz Polat

Received Date: 21.04.2022 Accepted Date: 13.04.2023 J Ankara Univ Fac Med 2023;76(2):122-127


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.


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.


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

Full Text (Turkish)