Is there a relationship between abuse of alcohol and illicit drugs seen in hair results?

Combined use of alcohol and illicit drugs is a serious health and social problem. In this study, it was examined, whether a relationship between alcohol and drug abuse can be ascertained by comparison of alcohol marker and drug concentrations in hair. In the frame of a social support system for families with parental abuse of illicit drugs, hair samples were analyzed between 2011 and 2022 for methadone, heroin (6-acetylmorphine), cocaine, amphetamine, ecstasy (MDMA), cannabinoids (THC), and the alcohol markers ethyl glucuronide (EtG) and ethyl palmitate (EtPa). For 1314 hair samples from adolescent and adult family members, the hair results show a prevalence of combined occasional or regular drug use and social or abusive alcohol use of 41%-60% except heroin (35%). The drug concentrations were statistically compared in the three categories of abstinence or moderate drinking, social drinking, and alcohol abuse. For the most frequently detected drug cocaine (n = 703), a significant increase of the concentrations with rising alcohol consumption was found. The frequent detection of cocaethylene proved the preferred simultaneous intake of both substances. For THC (n = 489), no significant difference between the alcohol consumer groups was seen. Concerning the less frequently detected methadone (n = 89), 6-acetylmorphine (n = 92), amphetamine (n = 123), and MDMA (n = 105), no clear trend between drug and alcohol marker results was determined. It is concluded that the evaluation of hair results is an appropriate way to study the extent of combined drug-alcohol consumption and complements other studies based on acquisition of consumption data by interview or questionnaire.

Repeated poisoning of the life partner by thallium - a case of questionable Munchausen by adult proxy syndrome with ensuing attempted murder.

After the use of thallium as rat poison was banned, the knowledge about the severe and treacherous course of poisonings with this toxic metal has widely been lost. In the present case, the male victim sustained two insidious poisoning attacks in 2017 and 2020 by the perpetrator, his female life partner. In the first poisoning episode, he suffered from increasing heavy pain of the abdomen, stinging pain of both legs, persistent obstipation, hyperesthesia, and, after about 2 weeks, tuft-wise loss of hair as typical symptoms of the thallium poisoning. Within 7 weeks, he was successively examined in six hospitals with a wide variety of diagnostic methods, but a conclusive explanation of the complaints was not found. The possibility of a metal intoxication was then suggested by the perpetrator who privately arranged the analysis of a blood sample with the result of 175 µg/l thallium. Although a criminal poisoning was assumed, the perpetrator was not identified. After the victim left the perpetrator, she subtly executed a second poisoning attack with thallium sulfate (blood level 1230 µg/l after 1 day, urine level 4760 µg/l after 10 days, and hair concentrations 3.26-0.49 from proximal to distal in 9 segments). The perpetrator was sentenced to 10.5 years imprisonment for grievous bodily harm and attempted murder. Because of the behavior of the perpetrator, a Munchausen by proxy syndrome was discussed as a motivation of the first poisoning but was excluded by the psychiatric expert because of a missing antisocial personality disorder.

Prevalence of cathinones and other new psychoactive substances in hair of parents and children of families with known or suspected parental abuse of conventional illegal drugs.

BACKGROUND: Hair analysis of parents and their children was regularly used since 2011 as a diagnostic tool in a social support project for families with known or suspected abuse of conventional illegal drugs and revealed a high incidence of cocaine, cannabinoids, amphetamines, ecstasy and heroin. In this context, the prevalence of new psychoactive substances (NPS) in these families should be important for a realistic estimate of the situation. METHODS: The extracts of 1537 hair samples from 318 children (age 1-14 years), 44 adolescents and 611 adults, which were collected and tested for conventional drugs between June 2016 and April 2021 and frozen at -20 °C, were reanalyzed by a validated LC-MS/MS method (limits of quantitation 5-24 pg/mg) for 33 cathinones, 10 phenylethylamines, 5 piperazines including the antidepressant trazodone, 2 tryptamines, 9 designer benzodiazepines, 4 synthetic opioids and 4 ketamine-like substances including phencyclidine. RESULTS: Between one and up to five from 42 of these substances were detected in 227 samples (14.8%). The most frequently detected substances were benzedrone (62x), α-pyrrolidinovalerophenone (41x), N-ethylamphetamine (29x), dimethyltryptamine (13x) and pyrovalerone (11x). The quantification was possible only for 34 results of 15 drugs and the remaining majority of the results were unambiguously identified below LLOQ. The relative frequency of conventional drugs in the 227 NPS positive samples was higher than in all 1310 NPS negative samples for cocaine (69.6% vs. 56.0%), heroin (6-acetylmorphine 8.8% vs. 4.9%), amphetamine (16.3% vs. 7.7%) and MDMA (16.3% vs. 7.0%) but was similar for THC (38.3% vs. 36.3%) and benzodiazepines (1.8% vs. 1.7%). The high prevalence of N-ethylamphetamine can be explained as a byproduct of the illicit amphetamine synthesis from benzaldehyde and nitroethane rather than as a separate drug or as a combined metabolite of amphetamine and ethanol. The isolated appearance of 3-trifluoromethylphenylpiperazine in 9 hair samples collected in January 2017 can be caused either by its use as an NPS or by its formation as a metabolite of the medical drug flibanserin. The results were compared within 17 families whose members were tested at the same time and showed positive NPS results. The detected drugs agreed between both parents only in about half of the cases whereas the drugs found in children's hair was always detected also in hair of one or both parents. CONCLUSION: The re-testing of hair extracts for NPS after long-time storage in frozen state enables an impression about the relative high prevalence in the tested population group, despite the limitation by partial degradation of the substances and the corresponding impossibility in quantitative assessments. In addition to conventional drugs, the hair test for these substances should be useful in unclear cases of child's welfare endangerment and in family law.

Positive findings of ethyl glucuronide in hair of young children from families with addiction background.

AIMS: Small children are expected to be abstinent from alcohol, and children's hair is frequently used as the blank matrix for calibration of the alcohol consumption marker ethyl glucuronide (EtG). The basal EtG concentrations of total abstainers were described to be 0.3-2.1 pg/mg (Pirro et al. 2013). It is examined whether this assumption is valid for children from families with addiction background. METHODS: In a social support system for families with drug and/or alcohol addicted parents, 161 hair samples from 126 children (age 1-14 years, hair segment 0-3 cm) were analyzed for EtG by a validated LC-MS/MS method (LOD 0.56 pg/mg, LLOQ 2.3 pg/mg). For comparison, ethyl palmitate (EtPa) was measured and hair samples from parents were included. EtG ≥ 3 pg/mg was considered as an alarming result for children. RESULTS AND DISCUSSION: EtG concentrations between 3.0 and 42.6 pg/mg (mean 9.55 pg/mg, median 6.40 pg/mg) were measured for 25 samples (15.5%, age 22 × 1-5 years, 3 × 9-11 years). Elevated EtPa (0.15-0.46 ng/mg) was found in 6 samples and cocaethylene (0.02-0.07 ng/mg) was detected in 5 samples with high cocaine findings. Hair results of one or both parents indicated drug abuse in 12 from 14 cases (85.7%) if both parents were tested. CONCLUSION: Although accidental or voluntary intake of alcoholic beverages cannot be excluded, the external contamination of children's hair by EtG-containing wine and sweat or urine of the alcohol abusing parents is assumed to be the most probable explanation for the positive EtG results in hair of 1-5-year-old children.

Hair analysis of more than 140 families with drug consuming parents. Comparison between hair results from adults and their children.

BACKGROUND: Hair samples from children are frequently analyzed in order to characterize their endangerment in a drug using environment. However, the interpretation of the results remains difficult because of lacking data for comparison. In this study, hair samples from families with drug consuming parents were analyzed for illegal and selected medical drugs and the results were evaluated concerning a relationship between findings of parents and children depending on kind of drug, age and gender of children as well as maternal or paternal drug concentrations in hair. METHODS: In an ongoing social supporting project for families with underage children and drug consuming parents, hair samples were analyzed since 2011 for methadone, opiates and opioid analgesics, cocaine, amphetamines, ecstasy, cannabinoids and benzodiazepines by LC-MS/MS with LOQs ≈ 0.01 ng/mg. From the data pool of more than 1300 individuals, 100 families with results for one or both parents and one to five children, 30 families with results only for both parents, and 11 families with results only for 2-4 children were selected. Fifty eight of these 141 families were repeatedly tested (altogether 251 family tests). RESULTS: One to 5 drugs were detected in 239 (95.2%) of the family tests with highest occurrence of cocaine (79.7%) and THC (50.2%). According to the concentrations of the tested persons, the most probable drug users were the mother (25%), the father (24%), both parents (16%), or were not tested (30%). Within the families, there was an agreement of the detected drugs between parents and children of 47.8%, between both parents of 36.1%, and between children of 42.3%. For parents with hair concentrations in the typical range of regular drug use, the drug was detected in children hair with the following frequency: methadone 65.5%, heroin (6-AM) 63.6%, cocaine 92.1%, amphetamine 80%, MDMA 42.9% and THC 67.4% with higher percentage for younger children. The agreement for medical drugs (benzodiazepines 7.7%, synthetic opioids 8.7%, diphenhydramine 7.1%) was much lower suggesting voluntary administration or intake. Despite the strong variation of the data, clear trends were found that the child/parent drug concentration ratio decreases with increasing children age and is higher for boys than for girls. CONCLUSION: The comparison of hair results within families gives a deeper insight in the drug situation, often enables the identification of the drug user and is helpful for social and legal decisions to improve the conditions of the children.

Commentary on current changes of the SoHT 2016 consensus on alcohol markers in hair and further background information.

The consensus on alcohol markers in hair was revised for the fourth time by an expert group of the Society of Hair Testing based on current state of research. This revision was adopted by the members of the Society during the business meeting in Brisbane on August 29th 2016. For both markers, ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs), two cut-off values for discrimination between teetotalers or occasional low amount consumption and moderate alcohol drinking (low cut-off), and between non-excessive (abstinence up to moderate alcohol intake) and chronic excessive drinking (high cut-off value) were critically examined. For the current revision, the cut-off values for EtG (7pg/mg and 30pg/mg, respectively) remained unchanged despite different findings or discussions published in the meantime. This was mainly due to the lack of broader data collections from new studies with great numbers of volunteers following thorough study concepts. In contrast, an essential change of the consensus was accepted for the FAEEs, where the concentration of ethyl palmitate (E16:0) can be used autonomously for interpretation instead of the concentration sum (ΣFAEE) of the four esters ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate, as previously applied. After evaluation of the data from seven laboratories, the E16:0 cut-off for abstinence assessment was defined at 0.12ng/mg for the 0-3cm segment and at 0.15ng/mg for the 0-6cm segment. The cut-off for chronic excessive drinking was fixed at 0.35ng/mg for the 0-3cm segment and at 0.45ng/mg for the 0-6cm segment. The use of E16:0 with these cut-offs in place of ΣFAEE for alcohol intake assessment produces only a minor loss in discrimination power, leads to no essential difference in the interpretation concerning chronic excessive alcohol consumption and is suitable to confirm EtG results in abstinence assessment if ethanol containing hair sprays or lotions are excluded.

High concentrations of lead and barium in hair of the rural population caused by water pollution in the Thar Jath oilfields in South Sudan.

In the oil fields of Thar Jath, South Sudan, increasing salinity of drinking water was observed together with human incompatibilities and rise in livestock mortalities. Hair analysis was used to characterize the toxic exposure of the population. Hair samples of volunteers from four communities with different distance from the center of the oil field (Koch 23km, n=24; Leer 50km, n=26; Nyal 110km, n=21; and Rumbek 220km, n=25) were analyzed for altogether 39 elements by inductively coupled plasma-mass spectrometry. Very high concentrations and a toxic health endangerment were assessed for lead and barium. The concentration of lead increased steadily with decreasing distance from the oil field from Rumbek (mean 2.8µg/g) to Koch (mean 18.7µg/g) and was there in the same range as in highly contaminated mining regions in Kosovo, China or Bolivia. The weighting materials in drilling muds barite (BaSO4) and galena (PbS) were considered to be the sources of drinking water pollution and high hair values. The high concentrations of lead and barium in hair demonstrate clearly the health risk caused by harmful deposition of toxic industrial waste but cannot be used for diagnosis of a chronic intoxication of the individuals.

Behaviour of hygrine and cuscohygrine in illicit cocaine production establishes their use as markers for chewing coca leaves in contrast with cocaine abuse.

Hygrine (HYG) and cuscohygrine (CUS) are natural alkaloids of coca leaves but are not found in illicit cocaine seizures. Therefore, they were proposed as markers for coca chewing in contrast to cocaine abuse in urine and hair testing. In order to examine at which step of the illegal cocaine production these compounds are lost, coca leaves were processed according to an authentic procedure by extraction with lime and kerosene, re-extraction with sulphuric acid, and precipitation of coca paste with ammonia. Non-extracted and extracted coca leaves, acidic extract and coca paste were analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) for cocaine, ecgonine methyl ester (EME), cinnamoylcocaine (CIN), HYG, and CUS. It follows from the results that under these conditions, HYG and CUS are extracted only to a minor extent by kerosene and are not precipitated from the acidic re-extract in the coca paste. Due to this behaviour in illegal cocaine production, they fulfil the conditions as markers for coca chewing in an optimal way. However, for unambiguous discrimination between coca chewing and cocaine abuse in human samples, additional markers of manufactured cocaine are required. Copyright © 2016 John Wiley & Sons, Ltd.

Drug resistance in cortical and hippocampal slices from resected tissue of epilepsy patients: no significant impact of p-glycoprotein and multidrug resistance-associated proteins.

Drug resistant patients undergoing epilepsy surgery have a good chance to become sensitive to anticonvulsant medication, suggesting that the resected brain tissue is responsible for drug resistance. Here, we address the question whether P-glycoprotein (Pgp) and multidrug resistance-associated proteins (MRPs) expressed in the resected tissue contribute to drug resistance in vitro. Effects of anti-epileptic drugs [carbamazepine (CBZ), sodium valproate, phenytoin] and two unspecific inhibitors of Pgp and MRPs [verapamil (VPM) and probenecid (PBN)] on seizure-like events (SLEs) induced in slices from 35 hippocampal and 35 temporal cortex specimens of altogether 51 patients (161 slices) were studied. Although in slice preparations the blood brain barrier is not functional, we found that SLEs predominantly persisted in the presence of anticonvulsant drugs (90%) and also in the presence of VPM and PBN (86%). Following subsequent co-administration of anti-epileptic drugs and drug transport inhibitors, SLEs continued in 63% of 143 slices. Drug sensitivity in slices was recognized either as transition to recurrent epileptiform transients (30%) or as suppression (7%), particularly by perfusion with CBZ in PBN containing solutions (43, 9%). Summarizing responses to co-administration from more than one slice per patient revealed that suppression of seizure-like activity in all slices was only observed in 7% of patients. Patients whose tissue was completely or partially sensitive (65%) presented with higher seizure frequencies than those with resistant tissue (35%). However, corresponding subgroups of patients do not differ with respect to expression rates of drug transporters. Our results imply that parenchymal MRPs and Pgp are not responsible for drug resistance in resected tissue.

Effect of the analyzed hair length on fatty acid ethyl ester (FAEE) concentrations in hair--is there congruence of cut-offs for 0-3 and 0-6 cm hair segments?

BACKGROUND: According to the current SoHT consensus for the use of alcohol markers in hair FAEEs can be analyzed in the proximal 0-3 or 0-6 cm segment with the cut-offs 0.2 and 0.4 ng/mg for abstinence assessment and 0.5 and 1.0 ng/mg for chronic excessive alcohol consumption. Both sets of cut-offs should be congruent for uniform interpretation. METHODS: FAEEs were determined in parallel for the 0-3 and the 0-6 cm segment of 157 hair samples and the concentration ratio between both segments (0-3 cm)/(0-6 cm) was calculated. For comparison, EtG was measured in the 0-3 cm segment of 135 of these samples and the FAEE concentration ratio was calculated by re-evaluation of segmental concentrations from further 42 samples of a previous study. RESULTS: The concentration ratio ranged from 0.3 to 1.5 (mean 0.83, median 0.82) and showed that the current cut-offs (ratio 0.50) are not congruent and that the cut-offs of the 0-3 cm are more restrictive against alcohol use or abuse. There was no correlation between the ratio and the concentration of FAEEs or EtG showing that the ratio does not depend on the drinking amount. Furthermore, no significant difference of the ratio was found between cosmetically untreated, bleached, dyed or spayed hair samples. Adaptation of the 0-3 cm cut-offs by increasing to 0.3 and 0.8 ng/mg respectively improved the congruence of both segment lengths but did not lead to a better agreement between FAEE and EtG interpretation. CONCLUSIONS: It follows from the results and further literature data that the variable length of the proximal hair segment between 3 and 6 cm which is possible according to the present SoHT consensus additionally contributes to the uncertainty of interpretation caused by biological variability and hair cosmetics. Furthermore, it is improbable from the present state of knowledge that a proximal segment length above 3 cm unambiguously enables the detection of alcohol consumption or abuse occurring more than 3 months before sampling. Therefore, it should be considered in a future consensus to redefine the hair length for both markers on 0-3 cm.

Possibilities for discrimination between chewing of coca leaves and abuse of cocaine by hair analysis including hygrine, cuscohygrine, cinnamoylcocaine and cocaine metabolite/cocaine ratios.

Contrary to the illegal use of any form of manufactured cocaine, chewing of coca leaves and drinking of coca tea are allowed and are very common and socially integrated in several South American countries. Because of this different legal state, an analytical method for discrimination between use of coca leaves and abuse of processed cocaine preparations is required. In this study, the applicability of hair analysis for this purpose was examined. Hair samples from 26 Argentinean coca chewers and 22 German cocaine users were analysed for cocaine (COC), norcocaine (NC), benzoylecgonine (BE), ecgonine methyl ester (EME), cocaethylene (CE), cinnamoylcocaine (CIN), tropacocaine (TRO), cuscohygrine (CUS) and hygrine (HYG) by hydrophilic interaction liquid chromatography (HILIC) in combination with triplequad mass spectrometry (MS/MS) and hybrid quadrupole time-of-flight mass spectrometry (QTOF-MS). The following concentrations (range, median, ng/mg) were determined in hair of the coca chewers: COC 0.085-75.5, 17.0; NC 0.03-1.15, 0.12; BE 0.046-35.5, 6.1; EME 0.014-6.0, 0.66; CE 0.00-13.8, 0.38; CIN 0.005-16.8, 0.79; TRO 0.02-0.16, 0.023; CUS 0.026-26.7, 0.31. In lack of a reference substance, only qualitative data were obtained for HYG, and two metabolites of CUS were detected which were not found in hair of the cocaine users. For interpretation, the concentrations of the metabolites and of the coca alkaloids in relation to cocaine were statistically compared between coca chewers and cocaine users. By analysis of variance (ANOVA) significant differences were found for all analytes (α = 0.000 to 0.030) with the exception of TRO (α = 0.218). The ratios CUS/COC, CIN/COC and EME/COC appeared to be the most suitable criteria for discrimination between both groups with the means and medians 5-fold to 10-fold higher for coca chewers and a low overlap of the ranges between both groups. The same was qualitatively found for HYG. However, these criteria cannot exclude cocaine use in addition to coca chewing. In this regard screening for typical cutting agents can be helpful and led to the detection of levamisole (21×), lidocaine (6×) and paracetamol (3×) in the 22 samples from German cocaine users, whereas no levamisole, lidocaine (3×) and paracetamol (1×) were found in hair from the Argentinean coca chewers. These criteria have to be confirmed for South American cocaine consumers including smokers of coca paste and may be different because of different composition of the drug and other use habits.

Cannabinoid findings in children hair - what do they really tell us? An assessment in the light of three different analytical methods with focus on interpretation of Δ9-tetrahydrocannabinolic acid A concentrations.

Hair analysis for drugs and drugs of abuse is increasingly applied in child protection cases. To determine the potential risk to a child living in a household where drugs are consumed, not only can the hair of the parents be analyzed but also the hair of the child. In the case of hair analysis for cannabinoids, the differentiation between external contamination and systemic uptake is particularly difficult, since the drug is quite often handled extensively prior to consumption (e.g. when preparing a joint) and smoke causes a further risk for an external contamination. Δ9-tetrahydrocannabinolic acid A (THCA-A), the non-psychoactive biogenetic precursor of Δ9-tetrahydrocannabinol (THC), is a suitable marker for external contamination since it is not incorporated into the hair matrix through the bloodstream in relevant amounts. In the presented study, hair samples from 41 children, 4 teenagers, and 34 drug-consuming parents were analyzed for THCA-A, THC and cannabinol (CBN) applying methanolic extraction and a fully validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method (Method 1). For comparison, a part of the samples was also analyzed applying alkaline hydrolysis followed by liquid/liquid extraction and gas chromatography-mass spectrometry (GC-M)S (Method 2), or by headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) (Method 3). Furthermore, 458 seized marihuana samples and 180 seized hashish samples were analyzed for the same cannabinoids by gas-chromatography-flame ionization detector (GC-FID). In all but one of the hair samples, the concentration of THCA-A was higher than the concentration of THC and in 14 cases no THC could be detected despite the presence of THCA-A, suggesting that in almost all cases a significant external contamination had occurred. Within-family comparison showed a higher THCA-A/THC ratio in hair of children than of their consuming caregivers. Mean and median of this ratio of all hair samples (6.7 and 4.2) were between those of marihuana (11.0 and 8.3) and hashish (2.8 and 2.1) with a large variation in all samples. Comparison of the Methods 1 to 3 showed clearly that the choice of the analytical procedure has a strong influence on the quantitative results, mainly because of decarboxylation of THCA-A during hair hydrolysis by NaOH and other analytical steps, which lead to artifactually elevated THC concentrations. In conclusion, these findings suggest that the major part of the cannabinoids detected in the hair samples from children arose from an external contamination through 'passive' transfer by e.g. contaminated hands or surfaces and not from inhalation or deposition of side stream smoke.

Meconium indicators of maternal alcohol abuse during pregnancy and association with patient characteristics.

AIM: Identification of women with moderate alcohol abuse during pregnancy is difficult. We correlated self-reported alcohol consumption during pregnancy and patient characteristics with objective alcohol indicators measured in fetal meconium. METHODS: A total of 557 women singleton births and available psychological tests, obstetric data and meconium samples were included in statistical analysis. Alcohol metabolites (fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG)), were determined from meconium and correlated with patient characteristics. RESULTS: We found that 21.2% of the 557 participants admitted low-to-moderate alcohol consumption during pregnancy. Of the parameters analyzed from meconium, only EtG showed an association with alcohol history (P < 0.01). This association was inverse in cases with EtG value above 120 ng/g. These values indicate women with most severe alcohol consumption, who obviously denied having consumed alcohol during pregnancy. No other associations between socioeconomic or psychological characteristics and the drinking status (via meconium alcohol metabolites) could be found. CONCLUSION: Women who drink higher doses of ethanol during pregnancy, according to metabolite measures in meconium, might be less likely to admit alcohol consumption. No profile of socioeconomic or psychological characteristics of those women positively tested via meconium could be established.

Methadone and illegal drugs in hair from children with parents in maintenance treatment or suspected for drug abuse in a German community.

BACKGROUND: Children living in homes with drug-addicted parents are in a steady danger of poisoning and may suffer from neglect, maltreatment, and lagging behind in development. Hair analysis could be a suitable way to examine this endangering exposure to drugs. METHODS: Hair samples from 149 children (aged 1-14 years) living with parents substituted by methadone and/or suspected for abuse of illegal drugs, and from 124 of the parents in a German community were investigated by liquid chromatography-hybrid quadrupole time-of flight mass spectrometry and by headspace solid phase microextraction gas chromatography-mass spectrometry for methadone, heroin, cocaine, amphetamines, ecstasy, cannabinoids and benzodiazepines and their metabolites or degradation products (32 compounds). RESULTS: From the children's hair, only in 35 samples, no drugs were detected. Cannabinoids were found in 56 samples, in 20 of them as the only drug. In the remaining 95 samples, methadone was identified 35 times with additional use of illegal drugs in 28 cases. Drug use in the children's environment was obvious for heroin in 44 cases, cocaine in 73 cases, amphetamine or ecstasy in 6 cases, and diazepam in 8 cases. The concentrations varied from limit of quantification to 2.16 ng/mg of methadone, 11.1 ng/mg of 6-acetylmorphine, 17.8 ng/mg of cocaine, 3.29 ng/mg of amphetamine, and 0.72 ng/mg of Δ-tetrahydrocannabinol. In general, hair from younger children contained higher concentrations than from their elder siblings. Systemic incorporation of methadone, cocaine, or cannabinoids appeared likely from detection of the nonhydrolytic metabolites 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine in 11 cases, norcocaine in 16 cases, and 11-nor-9-carboxy-Δ-tetrahydrocannabinol in 9 cases. Within the families, hair samples of children and parents provided often the same drug pattern. External deposition from smoke and by contact with contaminated surfaces or parent's hands and systemic deposition after passive smoking, administration, or oral intake by hand-to-mouth transfer were discussed as alternative incorporation mechanisms into hair. CONCLUSIONS: Altogether, investigation of children's hair proved to be a useful way to detect endangering drug use in their environment and lead to a more thorough inspection and measures to improve their situation in many of the cases.

Ethyl glucuronide concentrations in beard hair after a single alcohol dose: evidence for incorporation in hair root.

BACKGROUND: Despite the growing importance of ethyl glucuronide (EtG) in hair for detection of chronic excessive alcohol consumption, the mechanism of incorporation is not yet clear. Deposition from sweat is believed to be the main route. In order to get more information, EtG was determined in daily shaved beard hair after single higher alcohol doses. METHODS: Three volunteers drank within 5.5 h 153, 165 and 200 g ethanol followed by abstinence. Daily shaved beard hair was analysed for EtG using a validated liquid chromatography-tandem mass spectrometry method with a limit of quantification of 2 pg/mg. RESULTS: For all three volunteers, small concentrations of EtG were already detected 9 h after end of drinking. The concentrations increased to maxima of 182, 242 and 74 pg/mg on days 2 to 4 and then gradually decreased to limit of quantification on days 8 to 10. DISCUSSION: The time course of EtG is discussed based on literature data about anatomic dimensions of the hair root, physiology of hair growth, kinetics of EtG formation and elimination in blood, and in comparison to literature results about drugs in beard hair. It follows that for beard hair the predominant portion of EtG is incorporated in the upper part of the hair root between suprabulbar region and isthmus leading to a positive zone of about 3 mm (8-9 days) after a single drinking event. Deposition from sweat which is only possible into the residual hair stubble after shaving and in the infundibulum down to the sebaceous gland mouth was found to be of minor importance but could play a greater role in long hair. CONCLUSION: It is concluded that EtG in hair fulfils the prerequisites for time-resolved interpretation of segmental concentrations and that a single excessive drinking can be well detected in sufficiently short hair segments. However, in the routinely investigated 3-cm proximal scalp hair segment and using the cutoff of 7 pg/mg, a negative result can be expected with high probability because of dilution by negative hair.

Isomerization of cannabidiol and Δ9-tetrahydrocannabinol during positive electrospray ionization. In-source hydrogen/deuterium exchange experiments by flow injection hybrid quadrupole-time-of-flight mass spectrometry.

RATIONALE: Liquid chromatography/tandem mass spectrometry (LC/MS/MS) is frequently used for analysis of cannabinoids in drug abuse control. Despite differences in structure, the isomers Δ(9) -tetrahydrocannabinol (THC) and cannabidiol (CBD) provide identical fragment spectra after positive electrospray ionization (ESI). For elucidation of the reason, hydrogen/deuterium (H/D) exchange experiments were performed. METHODS: Solutions of THC and CBD in D(2) O/acetonitrile (50:50, v/v) were flow-injected into acetonitrile as the mobile phase and measured by hybrid quadrupole-time-of-flight mass spectrometry (FI-QTOF-MS) in targeted MS/MS mode. The MS and collision-induced dissociation (CID) spectra at 10, 20 and 40 eV were interpreted with respect to number and position of exchanged hydrogen atoms. For comparison the same measurements were preformed in H(2) O, after addition of 0.5% formic acid and with negative ESI. RESULTS: Depending on injected volume and position in the response curve, up to 7 or 8 hydrogen atoms were exchanged by deuterium in THC or CBD. Positive ESI CID spectra were available for precursors with up to 4 exchanged D-atoms and showed that besides the OH groups also an H/D exchange at carbon atoms of the non-aromatic part of the molecules occurred for both THC and CBD. After negative ESI, no H/D exchange in addition to the OH groups and different CID spectra of both substances was found. CONCLUSIONS: Injection of the investigated substances in D(2) O and measurement by FI-QTOF-MS proved to be an efficient way to perform H/D exchange experiments. The results were interpreted as an acid-catalyzed in-source equilibration between THC and CBD leading to the same precursor ions and to an H/D exchange in the methyl groups under the increased acidic conditions in the positive ESI droplets. Therefore, in positive LC/ESI-MS/MS, peak identification by CID spectra or by abundance ratio of multiple reaction monitoring (MRM) transitions is not sufficient for unambiguous discrimination between THC and CBD and must be supported by retention time or other experimental evidence.

General unknown screening in hair by liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-QTOF-MS).

The retrospective investigation of the exposure to toxic substances by general unknown screening of hair is still a difficult task because of the large number of possible poisons, the low sample amount and the difficult sample matrix. In this study the use of liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) was tested as a promising technique for this purpose. In the optimized procedure, 20mg hair were decontaminated with water and acetone and two times extracted by 18h incubation with 0.5ml of a mixture of methanol/acetonitrile/H(2)O/ammonium formate at 37°C. A mixture of deuterated standards from different drug groups was added for quantification and method control. The united extracts were evaporated to a residue of 0.5ml and 5µl were injected without clean-up for LC-QTOF-MS measurement (instrument Agilent 6530) with positive electrospray ionization and in data dependent acquisition mode. For peak identification the accurate mass data base and spectral library of the authors was used which contains accurate mass CID spectra of more than 2500 and theoretically calculated accurate mass data of more than 7500 toxicologically relevant substances. Validation at the example of 24 illegal drugs, their metabolites and benzodiazepines resulted in limits of detection of 0.003-0.015ng/mg, and limits of quantification of 0.006-0.021ng/mg with good accuracy and intra- and interday reproducibility. The matrix effect by ion suppression/enhancement was 72-107% for basic drugs and 42-75% for benzodiazepines. Yields of the hair extraction above 90% were determined for 59 drugs or metabolites. The method was applied to hair samples from 30 drug fatalities and from 60 death cases with known therapeutic drug intake at life time. Altogether 212 substances were identified with a frequency per drug of 1-40 (mean 4.2) and per case of 2-33 (mean 10.2), between them 35 illegal drug related substances and 154 therapeutic drugs. Comparison with the data known from case histories and from the analysis of blood, urine and gastric content showed only a low agreement, with many unexpected drugs detected and many reported drugs not detected in hair. Basic drugs and metabolites such as opioides, cocaine, amphetamines, several groups of antidepressants, neuroleptics, beta-blockers or the metamizole metabolite noramidopyrine were found with high frequency whereas acidic and several neutral drugs such as cannabinoids, salicylic acid, furosemide, barbiturates, phenprocoumone or cardiac glycosides could not be detected with sufficient sensitivity, mainly because of the low ion yield of positive ESI for these compounds. The advantage of a comprehensive acquisition of all substances is paid by a lower sensitivity in comparison to targeted screening LC-MS/MS procedures. In conclusion, the procedure of sample preparation and LC-QTOF-MS analysis proved to be a robust and sensitive routine method in which the qualitative screening for a wide variety of toxic substances in hair is combined with the quantitative determination of selected illegal drugs.

Interpretation problems in a forensic case of abstinence determination using alcohol markers in hair.

In a child custody case a mother with a longstanding history of alcohol misuse had to show absolute abstinence for one year. She entered a residential rehabilitation for six months and was tested two months later by way of a hair test for ethyl glucuronide (EtG) with the result of 22 pg/mg in the proximal 0-1cm segment and the segments 1-2 cm and 2-3 cm being negative. This was interpreted as a minimum alcohol intake of 20-50 units per week in the month before sampling. Since the mother denied any alcohol intake a second hair sample was collected seven weeks after the first and analyzed for fatty acid ethyl esters (FAEEs) by a second laboratory. A low concentration of 0.03 ng/mg was measured within the 0-6 cm segment of recently bleached hair and was interpreted as showing no evidence of alcohol use during the last six months. Three further hair samples were analyzed during the next nine months with low EtG values (<2.4-3.3 pg/mg, 0-3 cm segment) and low FAEE values (0.27-0.53 ng/mg, 0-6 cm segment). These findings were summarized as indicating continued low alcohol consumption over the past one year period. As a consequence of the conflicting results, the case was dealt with in a hearing before the Family Division of the High Court of London. It was concluded in the judgment that the evidence did not indicate that the mother had consumed alcohol in the period tested by the hair samples. It was stated that the evidence in this case highlighted the need for the exercise of considerable caution when hair tests for alcohol are being interpreted and relied upon, both generally and particularly in isolation, and that this case is a proper reminder of the need for expert evidence to be given in a manner according to the Practice Direction.