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Development of a UPLC–MS/MS method for determining γ-hydroxybutyric acid (GHB) and GHB glucuronide concentrations in hair and application to forensic cases

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Abstract

We present a series of forensic cases measuring concentrations in hair of γ-hydroxybutyric acid (GHB) and its glucuronide. The compounds were extracted from hair by incubation for 1.5 h in a 25:25:50 (v/v/v) mixture of methanol/acetonitrile/2 mM ammonium formate (8 % acetonitrile, pH 5.3). The compounds were determined by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). The limits of quantitation (LOQ) were 0.32 and 0.48 ng/mg hair for GHB and its glucuronide, respectively, and linearity up to 50 ng/mg. Extraction recoveries were 62–92 % for the analytes, and the accuracy was 90–108 %. Precision values expressed as relative standard deviations (%) obtained from daily controls were 9.1–11.3 % for GHB and 12.2–17.9 % for GHB glucuronide. Endogenous levels were up to 1.0 ng/mg for GHB and 1.2 ng/mg for GHB glucuronide in control subjects. In two GHB abusers, GHB was highly elevated (461–591 ng/mg), whereas GHB glucuronide was only slightly elevated in one of the subjects (1.7–3.1 ng/mg) and below the LOQ in the other. Thus, GHB glucuronide is not incorporated in hair in any appreciable amounts in GHB abusers and does not contribute diagnostically with regard to detection of exogenous exposure. To our knowledge, this is the first report to present GHB glucuronide in human hair.

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Acknowledgments

We appreciate Daniel Sejer Pedersen from Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen for providing reference standards of GHB glucuronide and its deuterium-labeled internal standard (GHB-d 4 glucuronide). We also thank the China Scholarship Council for providing financial support under the State Scholarship.

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Authors and Affiliations

  1. Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Frederik V’s Vej 11, 2100, Copenhagen, Denmark

    Xin Wang, Kristian Linnet & Sys Stybe Johansen

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  1. Xin Wang
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  2. Kristian Linnet
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Correspondence to Xin Wang.

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Informed consent was obtained from all healthy individuals included in the study, who supplied small strands of scalp hair for use as control materials.

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Wang, X., Linnet, K. & Johansen, S.S. Development of a UPLC–MS/MS method for determining γ-hydroxybutyric acid (GHB) and GHB glucuronide concentrations in hair and application to forensic cases. Forensic Toxicol 34, 51–60 (2016). https://doi.org/10.1007/s11419-015-0285-6

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  • DOI: https://doi.org/10.1007/s11419-015-0285-6

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