Date of Award

Spring 5-31-2022

Document Type

Thesis

Degree Name

Master of Science (MS)

Department/Program

Forensic Science

Language

English

First Advisor or Mentor

Richard Stripp

Second Reader

Teeshavi Acosta

Third Advisor

Damon Borg

Abstract

An ELISA method was developed and validated to detect ketamine in human hair samples. Ketamine is an anesthetic drug that causes memory loss, dissociative sensations, and hallucinations. Due to these adverse effects, ketamine is a common drug used in drug facilitated sexual assaults (DFSAs). It is very important to be able to detect the substances used in DFSAs over a longer period of time due to the delayed reporting of these crimes. Victims, often out of fear and from the sedative/memory loss effects of the drugs, tend to report these crimes when it is too late to use urine and blood for toxicological testing. Hair has a window of detection of up to 12 months, which makes it a useful matrix to use in DFSAs. The linear range of this assay was from 0 pg/mg to 1,000 pg/mg with a calibration curve returning an acceptable R2 value of 0.9991. The lower limit of detection was calculated to be 18.1 pg/mg. Accuracy and precision of this assay was determined through replicate analysis of quality control samples. Both intraday (n = 12) and interday (n = 12) accuracy and precision data were within the acceptable limits of +/- 20% error and 10% CV. Results indicated interference with PCP, which generated a response similar to a low positive control. There was no carryover seen between samples in the wells from the plate washer or from manual pipetting. This validated method was used to analyze positive authentic hair samples from donors with reported ketamine drug use. Results indicated correlation between the ELISA screening results compared to the LC-MSMS confirmation results. However, more samples need to be tested for further research. After thorough analysis, the Ketamine Direct ELISA kit from Immunalysis is suitable to use as a screening assay to detect ketamine in hair samples.

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