Significance of Metabolite Ratios in the Interpretation of Segmental Hair Testing Results—Differentiation of Single from Chronic Morphine Use in a Case Series

In morphine intoxication cases, forensic toxicologists are frequently confronted with the question of if the individual was opioid-tolerant or opioid-naïve, which can be investigated by hair analysis. However, interpretation of results can be challenging. Here, we report on hair testing for morphine and its metabolite hydromorphone following morphine intoxication without tolerance and upon chronic use. Two consecutive hair samples were collected after a non-fatal intoxication. Analysis comprised short hair segments and their initial wash water solutions. In the intoxications, morphine and hydromorphone levels were 3.3 to 56 pg/mg and at maximum 9.8 pg/mg, respectively. Both levels and hydromorphone to morphine ratios were significantly lower compared to chronic morphine use. In the non-fatal intoxication, the highest hydromorphone to morphine ratio was obtained in the segment corresponding to the time of intoxication. Morphine ratios of wash to hair were significantly higher in the intoxications compared to chronic use, being indicative of sweat/sebum contamination. We recommend including the analysis of hydromorphone and the initial wash solution in cases of morphine intoxications. Our study demonstrates that hydromorphone to morphine ratios can help in distinguishing single from chronic morphine use and in estimating the period of exposure when a consecutive hair sample can be collected in survived intoxications.

Hair toxicology identifies greater substance use than self-report in high-risk 9-13 year-olds in the ABCD Study

Aim: A key aim of the Adolescent Brain Cognitive Development (ABCD) Study is to document substance use onset, patterns, and sequelae across adolescent development. However, substance use misreporting obscures accurate drug use characterization. Hair toxicology tests provide objective historical substance use data, but are rarely investigated in youth. Here, we compare objective hair toxicology results with self-reported substance use in youth. Methods: A literature-based substance use risk algorithm identified 696 ABCD Study participants for hair sample collections between baseline and 2-year follow-up (spanning ages 9-13) for laboratory analysis. Chi-square and t-tests assessed differences between participants demographics, positive and negative hair tests, risk algorithm scores, and self-reported substance use. Results: Hair testing confirmed that 17% of at-risk 9-13 year-olds had evidence of past 3-month use of one (n=99), two (n=17), three (n=3), or four (n=2) drug classes. After considering prescribed medication use, 11% had a positive test incongruent with self- or parent-report. No participant with a positive result self-reported substance use consistent with their toxicology results. Participants with positive tests under-reported use (p<.001), reported less sipping of alcohol (p<.001), and scored higher on the risk algorithm (p<.001) than those with negative hair toxicology. Conclusions: An alarming 11% of tested samples in at-risk 9-13 year-olds were positive for at least one unreported substance, suggesting underreporting in this population when participating in a national healthy development study. The degree of underreporting cannot yet be calculated, as at-risk samples were prioritized for assays. Expanded toxicology testing is key to characterize substance use in youth.

In a Case of Death Involving Steroids, Hair Testing is more Informative than Blood or Urine Testing

A 59-year old male was found dead at home, with 2 empty vials of an oily preparation attributed to a manufacturer from East Europe. There was no label on the vial. The subject was a former weightlifter, also known as an anabolic steroids abuser. The local prosecutor ordered a body examination, which was unremarkable and allowed collecting femoral blood, urine and scalp hair (6 cm, brown). He was treated for cardiac insufficiency with quinidine. Biological specimens were submitted to standard toxicological analyses including a screening with LC-QToF, but also to a specific LC-MS/MS method for anabolic steroids testing. Ethanol was not found in both blood and urine. Quinidine blood concentration (791 ng/mL) was therapeutic. No drug of abuse was identified. In blood, testosterone was less that 1 ng/mL and no other steroid was identified. In urine, T/E was 1.56 and boldenone showed up at 9 ng/mL. The hair test results, performed on the whole length, demonstrated repetitive steroids abuse, including testosterone (140 pg/mg), testosterone propionate (605 pg/mg) and testosterone decanoate (249 pg/mg), but also boldenone (160 pg/mg), trenbolone (143 pg/mg) and metandienone (60 pg/mg). Given forensic laboratories have limited access to steroid urinary metabolites reference material due to specific regulations (to avoid testing athletes before anti-doping verifications), hair analyses seem to be the best approach to document anabolic agents abuse. Indeed, in hair, the target drug is the parent compound and, in addition, when compared to blood or urine, this matrix has a much larger window of detection. The pathologist concluded to cardiac insufficiency in a context involving repetitive abuse of anabolic drugs. This case indicates that more attention should be paid to anabolic steroids, in a context of sudden cardiac death.

Performance of self-reported measures of alcohol use and of harmful drinking patterns against ethyl glucuronide hair testing among young Swiss men

Background There is a need for empirical studies assessing the psychometric properties of self-reported alcohol use as measures of excessive chronic drinking (ECD) compared to those of objective measures, such as ethyl glucuronide (EtG). Objectives To test the quality of self-reported measures of alcohol use and of risky single-occasion drinking (RSOD) to detect ECD assessed by EtG. Methods A total of 227 samples of hair from young Swiss men were used for the determination of EtG. Self-reported measures of alcohol use (previous twelve-month and previous-week alcohol use) and RSOD were assessed. Using EtG (<30 pg/mg) as the gold standard of ECD assessment, the sensitivity and specificity were computed, and the AUROC were compared for alcohol use measures and RSOD. Logistic regressions were used to test the contribution of RSOD to the understanding of ECD after controlling for alcohol use. Results A total of 23.3% of participants presented with ECD. Previous twelve-month alcohol use with a cut-off of >15 drinks per week (sensitivity = 75.5%, specificity = 78.7%) and weekly RSOD (sensitivity = 75.5%, specificity = 70.1%) yielded acceptable psychometric properties. No cut-off for previous-week alcohol use gave acceptable results. In the multivariate logistic regression, after controlling for the previous twelve months of alcohol use, RSOD was still significantly associated with EtG (p = .016). Conclusion Self-reported measures of the previous twelve months of alcohol use and RSOD were acceptable measures of ECD for population-based screening. Self-reported RSOD appeared to be an interesting screening measure, in addition to the previous twelve months of alcohol use, to understand ECD among young people.

Analysis of Cocaine and Its Metabolites in Urine after Consumption of Coca Tea by Five Subjects and Subsequent Hair Testing

Coca tea is a popular drink in some countries of South America where it is reputed to have medicinal properties. This preparation is composed of natural cocaine (COC) alkaloids and therefore can be banned in some countries. During an anti-doping control in Peru, the urine of an athlete was tested positive for benzoylecgonine (BZE), ecgonine methyl ester (EME) and COC (400, 180 and 0.5 ng/mL, respectively). The athlete indicated that she had consumed coca tea in the morning before the competition. As her lawyer contacted us to assess the scientific aspects of the possible involvement of coca tea to explain the adverse analytical finding, a study was implemented with similar tea bags. Five volunteers from the laboratory consumed 250 mL of coca tea containing approximately 3.8 mg of COC. Eleven urine specimens were collected for each subject for 3 days to follow the elimination of COC and metabolites (BZE and EME). All samples were analyzed by UHPLC-MS/MS (Ultra High Performance Liquid Chromatography - tandem mass spectrometer) after alkaline extraction. Maximum detection times for COC was 20 h, with concentrations ranging from 6 to 91 ng/mL. Maximum detection times for BZE and EME were 70 h and 60 h, respectively, with concentrations ranging from 6 to 3,730 ng/mL and from 6 to 1,738 ng/mL. The concentration profiles were identical for the five volunteers. This study supports the athlete’s claims. In addition, the sample of hair strands of the five subjects was collected a month later and all the hair tests showed a negative result for COC with a limit of decision of 10 pg/mg. Although it is accepted that a 4-mg dose of COC has no significant pharmacological effect, the consummation of coca tea can lead to significant legal consequences since the measured urine concentrations sometimes cannot be considered incidental. Therefore, discrimination between coca tea consummation and recreational COC abuse relies primarily on hair analysis.