Heroin-Related Compounds and Metabolic Ratios in Postmortem Samples Using LC–MS-MS

Abstract

Analysis of postmortem samples with the presence of morphine can sometimes be challenging to interpret. Tolerance complicates interpretation of intoxications and causes of death due to overlap in therapeutic and fatal concentrations. Determination of metabolites and metabolic ratios can potentially differentiate between abstinence, continuous administration, and perhaps time of administration. The purpose of this study was to (a) develop and validate a method for quantitation of morphine-3β-D-glucuronide, morphine-6β-D-glucuronide, normorphine, codeine-6β-D-glucuronide, norcodeine, codeine, 6-acetylmorphine, and ethylmorphine in urine using liquid chromatography–tandem mass spectrometry; (b) apply the method to opiate related deaths; (c) compare metabolic ratios in urine in different causes of death (CoD) and after different drug intakes and (d) compare heroin intoxications in rapid and delayed deaths. Validation parameters such as precision, bias, matrix effects, stability, process efficiency, and dilution integrity were assessed and deemed acceptable. Lower limits of quantitation ranged from 0.01–0.2 μg/mL for all analytes. Autopsy cases (n=135) with paired blood and urine samples were analyzed. Cases were divided into three groups based on CoD; opiate intoxication, intoxication with other drugs than opiates, and other CoD. The cases were classified by intake: codeine (n=42), heroin (n=36), morphine (n=49), and ethylmorphine (n=3). Five cases were classified as mixed intakes and excluded. Heroin intoxications (n=35) were divided into rapid (n=15) or delayed (n=20) deaths. Parent drug groups were compared using metabolic ratio morphine-3β-D-glucuronide/morphine and significant differences were observed between codeine vs morphine (p=0.005) and codeine vs heroin (p≤0.0001). Urine and blood concentrations, and metabolic ratios in rapid and delayed heroin intoxications were compared and determined a significant difference for morphine (p=0.001), codeine (p=0.009), 6-acetylmorphine (p=0.02) in urine, and morphine (p=0.02) in blood, but there was no significant difference (p=0.9) between metabolic ratios. Morphine-3β-D-glucuronide results suggested a period of abstinence prior to death in 25% of the heroin intoxications.

Oxycodone in Forensic Toxicology : Analytical Strategies and Interpretation

“…This has been described for e.g. buprenorphine (26), and heroin (27,28). A negative opioid result in urine or hair can suggest short-term abstinence or long-term abstinence which could possibly explain the events leading to death.…”

Metabolic Evaluation of Synthetic Opioids on the Example of U-47700 with the Use of In Vitro and In Vivo Methods for Forensic Toxicology Application

“…The in vitro and in vivo study of the metabolic profile of NPS is a useful tool for providing data. This may be useful in terms of diagnosis, assessment of the severity of poisoning and treatment for the purposes of clinical toxicology, or the toxicological distinction of “rapid deaths” and “delayed deaths” [ 24 , 25 ], predicting possible post mortem redistribution processes and their impact on the final result of determinations for the purposes of medical and forensic opinion.…”

Heroin-Related Fatalities in Jeddah, Saudi Arabia, between 2008 and 2018

“…The level of codeine was low in all age groups (median, 20–35 ng/mL), which could be explained by the fact that 6-AC is metabolized to codeine and codeine-6-glucuronide (C6G) faster than to other compounds. A recently published study demonstrated that C6G was not detected in blood samples obtained from authentic postmortem cases following heroin intake, while codeine was [ 42 ]. In another study, codeine was always higher than C6G in autopsy blood samples from heroin- and codeine-related fatalities [ 19 ].…”

“…The use of blood-free morphine/total morphine ratios in the blood to identify the cause and mode of death has been reported [ 18 , 42 ]. However, this approach always involves certain issues.…”

“…Consistent with previous reports, deaths following heroin intake can be divided into three types: rapid and delayed deaths, and undetermined survival times [ 2 , 3 , 51 ]. Rapid deaths can be identified by the detection of 6-MAM in blood samples, fresh needle marks, and the presence of syringes attached to the deceased body or nearby [ 3 , 42 , 59 ]. In addition, valuable information on the last moment before death can be obtained from family, friends, or witnesses [ 13 ].…”