Insights into the decontamination of cocaine-positive hair samples

A highly discussed step in hair sample preparation for forensic analytics is the applied decontamination. The here presented investigations aim to gain insight and give recommendations on how to conduct this decontamination for the analysis of cocaine consumption in hair. Key insights were gained from the investigation of cocaine consumer hair, which was artificially contaminated in a humid atmosphere with 13C6 labelled cocaine and from cocaine powder contaminated hair. Several decontamination protocols were investigated, whereby the usage of a decontamination protocol consisting of multiple short repetitive washes allowed to visualize the wash-out of (13C6-) cocaine. Multiple methanol washes proved to be an efficient and simple decontamination approach. Our findings showed that decontamination protocols can successfully wash-out recent cocaine contaminations. They were observed to be rather quickly washed-out, whereas cocaine from consumption or “older” cocaine contaminations were shown to eliminate both at a constant rate (from inner hair compartments). Thus, the usage of decontamination protocols to differentiate between consumption and contamination was shown to be limited. As contamination can happen any time at any level, only the application of elaborated decision trees, based on cocaine metabolite ratios and thresholds, can provide the distinction between consumption and contamination. Thus, the authors highly recommend the usage of such tools on all hair samples analyzed for cocaine consumption

Single sample preparation for the simultaneous extraction of drugs, pharmaceuticals, cannabinoids and endogenous steroids in hair

A combined sample preparation for the simultaneous extraction of drugs, pharmaceuticals, cannabinoids and endogenous steroids was developed based on three independent fully validated analytical methods. Recently, we published a multi-analyte method for the simultaneous analysis of 116 drugs and pharmaceuticals including different substance groups like opioids, stimulants, benzodiazepines, z-drugs, antidepressants and neuroleptics based on a single sample workup followed by a single analytical measurement with LC-MS/MS. However, in some cases, additional analysis of further substance groups, such as cannabinoids and endogenous steroids, is required, which are analyzed in our laboratory using separate sample preparation and separate analytical methods. The goal of this study was to use the knowledge from the different sample preparations and combine them into a single sample preparation and extraction workflow for the simultaneous extraction of drugs, pharmaceuticals, cannabinoids, and endogenous steroids to be analyzed with the appropriate analytical methods. A partial validation of selected parameters such as selectivity, linearity, limit of quantification (LOQ), accuracy, precision and robustness for the different analytical methods was carried out and revalidated. In addition, comparative measurements of quality controls and authentic pools were performed and statistically evaluated using the unpaired t-test or the non-parametric Mann-Whitney test. The results using the newly established sample preparation and extraction were in good agreement with the original data. In conclusion, the newly established sample preparation is suitable for the combined extraction of drugs, pharmaceuticals, cannabinoids and endogenous steroids, and gives reliable results for quantification of various substances

LC-MS/MS analysis of Δ9-THC, CBN and CBD in hair: investigation of artefacts

In forensic toxicology, high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) is increasingly used for the fast and sensitive measurement of a wide range of drugs. For our routine casework, a liquid chromatography atmospheric pressure chemical ionization-tandem mass spectrometry (LC-APCI-MS/MS) method for the quantification of Δ9-tetrahydrocannabinol (Δ9-THC), cannabinol (CBN) and cannabidiol (CBD) in hair was established and fully validated. Separation was achieved using a Kinetex® C18 column (100 mm × 2.1 mm, 100 Å, 1.7 μm, Phenomenex) at a flow rate of 0.5 mL/min. Measurements were performed on a QTrap 5,500 mass spectrometer (Sciex, Darmstadt, Germany). Unexpected signals were observed in authentic THC-positive hair samples. First, a signal with a slightly shifted retention time of THC whose origin could be assigned to the isomer Δ8-THC. Second, additional peaks exhibiting the same fragments as CBN and Δ9-THC but eluting at different retention times were detected. Spiking experiments and enhanced product ion scans (EPI) pointed to the origin of these additional signals as result of in-source decarboxylation of Δ9-tetrahydrocannabinolic acid A (Δ9-THCA-A) into Δ9-THC and further partial oxidation of Δ9-THC into CBN, respectively. Positive findings of Δ9-THCA-A in hair have been shown to derive from external contamination, therefore, the herein described artefacts may be used as indirect markers for external contamination

Simultaneous quantification of steroid hormones and endocannabinoids (ECs) in human hair using an automated supported liquid extraction (SLE) and LC-MS/MS – Insights into EC baseline values and correlation to steroid concentrations

Endogenous steroid hormones and endocannabinoids (ECs) are important regulators in the stress response of the human body. For the measurement of chronic stress, hair analysis has been established as method of choice for long-term and retrospective determination of endogenous stress markers. A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of five steroid hormones (cortisone, cortisol, androstenedione, testosterone, progesterone) and four endocannabinoids (anandamide, palmitoylethanolamide, 2-arachidonylglycerol, oleoylethanolamide) in hair was developed and validated. The hair samples were extracted with methanol and cleaned up with a fully automated supported liquid extraction (SLE) before analysis. Special attention was paid to the difficulties accompanying the quantification of endogenous analytes in hair. Five different strategies for endogenous compound quantification in hair (surrogate analyte, standard addition, background correction, stripped matrix and solvent calibration) were tested and compared. As a result, the approach of the surrogate analyte was used for the quantification of steroid hormones whereas background correction was used for endocannabinoids. The measurement of 58 samples from healthy young adults allowed insights into endocannabinoid ranges in hair and the correlation to steroid hormones. No significant differences in steroid and EC concentration levels of male and female in hair were found, except for testosterone (p < 0.001) and androstenedione (p < 0.0001). Cortisol to cortisone and testosterone to androstenedione concentrations were significantly and positively correlated. There were significant intercorrelations between endocannabinoids

Time-dependent postmortem redistribution of butyrfentanyl and its metabolites in blood and alternative matrices in a case of butyrfentanyl intoxication

A fatal case of butyrfentanyl poisoning was investigated at the Zurich Institute of Forensic Medicine. At admission at the institute approx. 9 h after death (first time point, t1), femoral and heart blood (right ventricle) was collected, as well as samples from the lung, liver, kidney, spleen, muscle and adipose tissue using computed tomography (CT)-guided biopsy sampling. At autopsy (t2), samples from the same body regions were collected manually. Additionally, urine, heart blood (left ventricle), gastric content, brain samples and hair were collected. Butyrfentanyl concentrations and relative concentrations of the metabolites carboxy-, hydroxy-, nor-, and desbutyrfentanyl were determined by LC⿿MS/MS and LC-QTOF. At t1, butyrfentanyl concentrations were 66 ng/mL in femoral blood, 39 ng/mL in heart blood, 110 ng/g in muscle, 57 ng/g in liver, 160 ng/g in kidney, 3100 ng/g in lung, 590 ng/g in spleen and 550 ng/g in adipose tissue. At t2, butyrfentanyl concentration in urine was 1100 ng/mL, in gastric content 2000 ng/mL, in hair 11,000 pg/mg and brain concentrations ranged between 200⿿340 ng/g. Carboxy- and hydroxybutyrfentanyl were identified as most abundant metabolites. Comparison of t1 and t2 showed a concentration increase of butyrfentanyl in femoral blood of 120%, in heart blood of 55% and a decrease in lung of 30% within 19 h. No clear concentration changes could be observed in the other matrices. Postmortem concentration changes were also observed for the metabolites. In conclusion, butyrfentanyl seems to be prone to postmortem redistribution processes and concentrations in forensic death cases should be interpreted with caution

In- and Out-Group Effects on Social Perception and Empathy in Cocaine Use Disorder

Earlier research revealed that cocaine users display impairments in emotional but not necessarily in cognitive empathy. However, no study to date has tested whether empathy is generally altered or whether impairments are restricted to specific social targets. The current investigation addresses this open question. In addition, we examined whether attributions of warmth and competence as well as personal future expectancies differed between cocaine users and substance-naïve controls. Twenty-two chronic cocaine users and 40 stimulant-naïve controls specified their perceived warmth and competence for four social targets [in-group member, opposite consumption out-group member (cocaine user for controls and non-user for cocaine user), opposite consumption out-group member of opposite gender, and elderly person]. They also specified their cognitive and emotional empathy for these four targets facing eight desirable and eight undesirable events. Finally, they rated the likelihood of these scenarios happening to themselves. Both cocaine users and controls attributed lower warmth to cocaine-using than non-using targets. Comparably, no in-group preference was observed in cocaine user’s emotional empathy ratings, and greater denigration of the in-group was associated with higher frequency and doses of cocaine consumption. In addition, cocaine users rated both desirable and undesirable events as more likely to happen to themselves than did controls. Results show that substance-naïve individuals stigmatize cocaine users. They further point to compromised self-esteem in cocaine users resulting from such stigmatization. Interventions should address stigmatization processes to break the vicious circle of mutual social distancing and stronger dedication to the drug

When Substance Use Is Underreported: Comparing Self-Reports and Hair Toxicology in an Urban Cohort of Young Adults

Objective: Large-scale epidemiological research often uses self-reports to determine the prevalence of illicit substance use. Self-reports may suffer from inaccurate reporting but can be verified with objective measures. This study examined the following: the prevalence of illicit and non-medical substance use with self-reports and hair toxicology, the convergence of self-reported and objectively quantified substance use, and the correlates of under- and overreporting. Method: The data came from a large urban cohort study of young adults (n = 1,002, mean age = 20.6 years, 50% female). The participants provided 3 cm of hair (covering the previous 3 months) and reported their illicit and non-medical substance use and their sociodemographic, psychological, and behavioral characteristics. Hair toxicology analyses targeted cannabinoids, ketamine, opiates/opioids, stimulants including 3,4-methylenedioxymethamphetamine, and relevant metabolites. Results: Self-reports underestimated the prevalence of most substances by 30% to 60% compared to hair tests. The average detection ratio (hair test/self-report) was 1.50. Hair tests were typically more sensitive than self-reports. Underreporting was associated with a low level of that substance in hair. Self-reported delinquency and psychopathology were correlated with an increased likelihood of concordant positive self-reports and hair tests compared to underreporting. Overreporting was associated with infrequent self-reported use. Conclusion: Our study suggests that self-reports underestimate young adults' exposure to illicit substances and non-medical use of prescription drugs. Consequently, estimates of associations between substance use and risk factors or outcomes are likely biased. Combining self-reports with hair tests may be most beneficial in study samples with occasional substance use. Researchers can use specific factors (eg, detection ratios) to adjust prevalence estimates and correlations based on self-reports

When Substance Use Is Underreported: Comparing Self-Reports and Hair Toxicology in an Urban Cohort of Young Adults

Objective: Large-scale epidemiological research often uses self-reports to determine the prevalence of illicit substance use. Self-reports may suffer from inaccurate reporting but can be verified with objective measures. This study examined the following: the prevalence of illicit and non-medical substance use with self-reports and hair toxicology, the convergence of self-reported and objectively quantified substance use, and the correlates of under- and overreporting. Method: The data came from a large urban cohort study of young adults (n = 1,002, mean age = 20.6 years, 50% female). The participants provided 3 cm of hair (covering the previous 3 months) and reported their illicit and non-medical substance use and their sociodemographic, psychological, and behavioral characteristics. Hair toxicology analyses targeted cannabinoids, ketamine, opiates/opioids, stimulants including 3,4-methylenedioxymethamphetamine, and relevant metabolites. Results: Self-reports underestimated the prevalence of most substances by 30% to 60% compared to hair tests. The average detection ratio (hair test/self-report) was 1.50. Hair tests were typically more sensitive than self-reports. Underreporting was associated with a low level of that substance in hair. Self-reported delinquency and psychopathology were correlated with an increased likelihood of concordant positive self-reports and hair tests compared to underreporting. Overreporting was associated with infrequent self-reported use. Conclusion: Our study suggests that self-reports underestimate young adults' exposure to illicit substances and non-medical use of prescription drugs. Consequently, estimates of associations between substance use and risk factors or outcomes are likely biased. Combining self-reports with hair tests may be most beneficial in study samples with occasional substance use. Researchers can use specific factors (eg, detection ratios) to adjust prevalence estimates and correlations based on self-reports