Defense Date
2022
Document Type
Directed Research Project
First Advisor
Dr. Michelle Peace
Second Advisor
Justin Poklis
Third Advisor
Dr. Sue Brown
Fourth Advisor
Lisa Davis
Abstract
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.
Rights
© The Author(s)
Is Part Of
VCU Master of Science in Forensic Science Directed Research Projects
Date of Submission
5-4-2022