A Retrospective Metabolomics Analysis of Gamma-Hydroxybutyrate in Humans: New Potential Markers and Changes in Metabolism Related to GHB Consumption

Tingting Wang; Kirstine L Nielsen; Kim Frisch; Johan K Lassen; Camilla B Nielsen; Charlotte U Andersen; Palle Villesen; Mette F Andreasen; Jørgen B Hasselstrøm; Mogens Johannsen

doi:10.3389/fphar.2022.816376

A Retrospective Metabolomics Analysis of Gamma-Hydroxybutyrate in Humans: New Potential Markers and Changes in Metabolism Related to GHB Consumption

Front Pharmacol. 2022 Mar 3:13:816376. doi: 10.3389/fphar.2022.816376. eCollection 2022.

Affiliations

¹ Department of Forensic Medicine, Section for Forensic Chemistry, Aarhus University, Aarhus, Denmark.
² Bioinformatics Research Centre, Aarhus University, Aarhus, Denmark.

Abstract

GHB is an endogenous short-chain organic acid presumably also widely applied as a rape and knock out drug in cases of drug-facilitated crimes or sexual assaults (DFSA). Due to the endogenous nature of GHB and its fast metabolism in vivo, the detection window of exogenous GHB is however narrow, making it challenging to prove use of GHB in DFSA cases. Alternative markers of GHB intake have recently appeared though none has hitherto been validated for forensic use. UHPLC-HRMS based screening of blood samples for drugs of abuse is routinely performed in several forensic laboratories which leaves an enormous amount of unexploited data. Recently we devised a novel metabolomics approach to use archived data from such routine screenings for elucidating both direct metabolites from exogenous compounds, but potentially also regulation of endogenous metabolism and metabolites. In this paper we used UHPLC-HRMS data acquired over a 6-year period from whole blood analysis of 51 drivers driving under the influence of GHB as well as a matched control group. The data were analyzed using a metabolomics approach applying a range of advanced analytical methods such as OPLS-DA, LASSO, random forest, and Pearson correlation to examine the data in depth and demonstrate the feasibility and potential power of the approach. This was done by initially detecting a range of potential biomarkers of GHB consumption, some that previously have been found in controlled GHB studies, as well as several new potential markers not hitherto known. Furthermore, we investigate the impact of GHB intake on human metabolism. In aggregate, we demonstrate the feasibility to extract meaningful information from archived data here exemplified using GHB cases. Hereby we hope to pave the way for more general use of the principle to elucidate human metabolites of e.g. new legal or illegal drugs as well as for applications in more global and large scale metabolomics studies in the future.

Keywords: Gamma-hydroxybutyrate; UPLC-QTOF analysis; biomaker discovery; driving under the influence of drugs (DUID); drug metabolism; metabolomics; retrospective study; whole blood samples.