Forensic toxicology is a branch of science that involves the analysis of drugs and other substances in biological fluids and tissues such as blood, urine, and oral fluid to aid medical or legal investigation of death, poisoning, and drug use. Due to the various components of different matrices, efficient and effective sample preparation techniques are necessary for reliable and accurate analysis. Following sample clean-up, a sensitive, specific, and robust method is ideal for consistent detection, identification, and quantitation of analytes. With the rise of drug abuse, there is a growing need to develop a single method that can target multiple classes of drugs quickly and effectively.
This study validated two different sample preparation techniques for the detection and quantitation of six drug classes comprised of twenty-three drugs and metabolites in oral fluid. The drug classes were as follows: amphetamines, local anesthetics, opioids, hallucinogens, antidepressants, and novel psychoactive substances (NPS). Amphetamines used were amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), and 3,4-methylenedioxymethamphetamine (MDMA). Local anesthetics contained benzoylecgonine (BZE), cocaine, and lidocaine. Opioids included codeine, methadone, morphine, 6-monoacetylmorphine (6-MAM), fentanyl, and oxycodone. Hallucinogens included lysergic acid diethylamide (LSD) and phencyclidine (PCP). Antidepressants were amitriptyline, citalopram, fluoxetine, and trazodone. Lastly, NPS included ethylone, α-pyrrolidinopentiophenone (α-PVP), and 2,5-dimethoxy-4-iodophenethylamine N-(2-methoxybenzyl) (25I-NBOMe). Supported liquid extraction (SLE) and solid phase extraction (SPE) were assessed followed by confirmatory analysis by liquid chromatography (LC)-tandem mass spectrometry (MS/MS).
Both methods were validated according to guidelines in the Standard Practices for Method Validation in Forensic Toxicology set by the American Academy of Forensic Science (AAFS) Standards Board (ASB). Parameters assessed include calibration model, bias, precision, limit of detection (LOD), limit of quantitation (LOQ), dilution integrity, ion suppression/enhancement, interference studies, and stability. Matrix recovery was added as another parameter. All calibration models were 0.99 or greater and all compounds were stable for at least 72 hours. Bias, precision, LOD, LOQ, dilution integrity, and interferences were similar between both methods. SLE yielded slightly better LOD and LOQ values. SLE had greater values of matrix recovery as well as lower levels of ionization suppression/enhancement.
Overall, SLE was determined to be the better method of sample preparation for this panel of drugs in oral fluid. Not only did it yield higher values for several of the parameters assessed but it also was more efficient (1 hour versus 2 hours) while using less solvent.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/41271 |
| Date | 08 July 2020 |
| Creators | Hwang, Hajin |
| Contributors | Botch-Jones, Sabra |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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