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Developing a Unified Method for the Analysis of Five Ethanol Metabolites: EtG, EtS, GTOL, 5-HTOL, and 5-HIAA using UPLC-MS/MS

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  • Title:
    Developing a Unified Method for the Analysis of Five Ethanol Metabolites: EtG, EtS, GTOL, 5-HTOL, and 5-HIAA using UPLC-MS/MS
  • Author: Jones, Bailey K
  • Subjects: EtG ; Ethanol ; EtS ; Forensic Science and Technology ; GTOL ; HIAA ; HTOL
  • Description: Ethanol analyses make up the largest category of quantitative assessments in forensic toxicology. Existing literature describes fit-for-purpose methodology for single or paired ethanol metabolites. The purpose of this study was to develop, and validate, a single method for the quantitation of ethyl glucuronide (EtG), ethyl sulfate (EtS), 5-hydroxytryptophol (5-HTOL), 5-hydroxytryptophol glucuronide (GTOL), and 5-hydroxyindole-3-aceteic acid (5-HIAA) in urine using UPLC-MS/MS. A single solid phase extraction method was performed using UCT Clean Screen FAST EtG SPE columns for analysis on a Shimadzu LC-MS/MS-8050. Chromatographic separation was performed on an Agilent InfinityLab Poroshell HPH-C18 column (4.6mm x 100mm x 2.7mm) at 55°C. The mobile phase was 0.1% formic acid in water (A) and acetonitrile (B). The limit of detection for EtG, EtS, GTOL, 5-HTOL, and 5-HIAA was determined to be 235, 29, 90, 4, and 13 ng/mL, respectively, and the lower limit of quantitation for EtG, EtS, GTOL, 5-HTOL, and 5-HIAA was determined to be 500, 100, 100, 10, and 20 ng/mL, respectively. Clinical samples from Paradigm Labs, LLC, were analyzed. EtG and EtS concentrations were evaluated above and below the clinical cut-off concentrations for 5-HTOL/5-HIAA and GTOL/5-HIAA. No statistically significant differences were observed between the population means for EtG concentrations below and above the clinical cutoff for 5-HTOL/5-HIAA. Statistically significant differences were observed between the population means for EtG and EtS concentrations below and above the clinical cutoff for GTOL/5-HIAA, as well as EtS concentrations below and above the clinical cutoff for 5-HTOL/5-HIAA.
  • Publisher: VCU Scholars Compass
  • Creation Date: 2022
  • Language: English
  • Source: VCU Scholars Compass
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