[Drinking study on the pharmacokinetics of the grappa congener 2-butanol].

A drinking study on the pharmacokinetics of the typical grappa congeners 2-butanol and 2-butanone (methyl ethyl ketone) was performed. It was expected that the concentration ratio might provide a means to estimate the time of ingestion of a grappa beverage. Twelve subjects drank a volume of the grappa "Vecchio di Prosecco" (42 vol%) to reach a blood alcohollevel of 1.20 %o. In the congener analyses in serum, a median 2-butanol concentration of 0.79 mg/1 (range 0.45-1.34 mg/1) and of 1.01 mg/I (0.44-1.62 mg/1) for 2-butanone were measured. The concentration-time curve was biphasic starting with a slow and plateau-like elimination. However, considerable inter-individual differences were observed. Only in 3 subjects, a 2-butanol : 2-butanone ratio below 1 suggested ingestion within the last 6 hours. The majority of the subjects exhibited higher concentrations of 2-butanone than of 2-butanol such that the ratio was always smaller than 1. According to the present results the concentrations of 2-butanol and 2-butanone or their ratio do not provide a reliable basis to draw conclusions on the time of grappa ingestion.

Field study to detect illicit and medicinal drugs in car drivers in Southern and Western Hesse.

In the present study, immunochemical tests (Mahsan DrugInspector, DOA4, DOA8, DOA10, Protzek) as well as the detection rate of police checks were evaluated. Urine and blood samples of suspected car drivers were analysed by chromatography-mass spectrometry. Additionally, anonymised urine samples were analysed on a voluntary basis in cases where no legal proceedings were initiated. Toxicological analyses (total unknown screening) were performed using gas chromatography-mass spectrometry (GC-MS) after hydrolysis, acidic and alkaline extraction and derivatization. A data base for screening 9000 substance entries was applied. In addition, urine samples were analysed using liquid chromatography/ time-of-flight mass spectrometry (HPLC-ToF-MS) to screen psychiatric and narcotic drugs. In total, samples of 154 suspects were analysed, of these, 46 samples for no actual reason. In 5 of the latter samples, forensically relevant substances were detected; in two cases the consumption of illicit drugs, i. e. cannabis and methamphetamine, was proved. Of the 154 suspects, 108 were charged with driving under the influence of drugs; in samples of 103 of these cases, illicit drugs were found. Immunochemical pretesting showed posi- tive results in 97 of the 108 cases; in 6 samples, psychiatric drugs (citalopram, doxepin, promethazine, mirtazapine, fluoxetine, venlafaxine) were later identified, which are not detectable by ordinary pretesting systems. Police officers successfully identified 95.4 % of the suspects as drug consumers, which is an excellent result. In practice, pretesting of urine samples using immunochemical techniques proved to be very reliable. The Protzek system in particular corresponded well with the results of the chromatographic analyses. In conclusion, systematic chromatographic-mass spectrometric analysis of urine samples of suspects is recommended to identify car drivers consuming illicit drugs and to obtain data usable in legal proceedings (e. g. suspending of the driving license), which is not always possible when using blood samples in cases of drugs consumed some time ago.

High variation of congener alcohols in apple wines.

Congener alcohol (CA) analysis became an important tool in forensic science to prove the kind of alcoholic beverage consumed. The aim of the present study was to determine the influence of yeast strain and apple variety on the formation of congener alcohols in self-produced apple wine. There exist data on CA patterns of industrially produced alcoholic beverages, but these are not available for apple wine. Must from five different commercial as well as from six genuine apple varieties were used for fermentation under similar conditions CA formation was monitored during the fermentation process. Additionally, nine commercial apple wines from commercial producers were analyzed. Analysis was performed by headspaces-GC-MS. All apple wines contained markedly high contents of the CA 3-methylbutan-1-ol (88-251 mg/L). Compared to self-produced apple wines from genuine musts the industrial apple wines (purchased in supermarkets and self-produced from commercial musts) exhibited significant differences in methanol concentrations(8.5-94 mg/L), whereas all other CAs, such as propan-1-ol, butan-1-0l, 2-methylpropan-1-ol(isobutanol), 3-methyl-butan-1-oi, and 2-methylbutan-1-oi, were found to be present in similar concentrations. Methanol was not detectable in apple wine made from genuine musts during fermentation but after a storage period. In some cases, concentrations of some CAs additionally changed during storage. This may be explained by a secondary (unwanted) fermentation after bottling. According to the data obtained in the present study, it is recommended to analyze a sample of the allegedly consumed apple wine in forensic cases, rather than to rely on data obtained from the literature or from some data collections.

Sleep self-intoxication and sleep driving as rare zolpidem-induced complex behaviour.

INTRODUCTION: The GABA(A) receptor agonist zolpidem has been used for treatment of insomnia since years, but special side effects have been reported. These side effects were called zolpidem-induced sleep-related complex behaviour. Such complex behaviour is associated with somnambulism and includes sleepwalking, sleep eating, sleep conversation and sleep driving. CASE PRESENTATION: Two cases of zolpidem-induced sleep-related complex behaviour following self-intoxication, sleep driving and amnesia are presented. In both cases, the subjects reported the voluntary intake of only one zolpidem tablet of 10 mg and amnesia for the time afterwards. Shortly after the onset of the drug's action, both individuals drifted into a somnambulism-like state and toxicological blood analysis suggested the intake of the remaining zolpidem tablets which might be called "sleep intoxication". Later, the subjects were arrested by police after driving under drug influence and not realizing the situation. Retrospectively, both subjects suffered from psychiatric disorders and in case 2, the subject was treated for depression with doxepin. Consequently, these co-factors may have increased the risk for the occurrence of the sleep-related complex behaviour. DISCUSSION: Involuntary self-intoxication should be taken into account in addition to the known pattern of zolpidem-induced complex behaviour. In legal cases, the forensic expert has to assess the blood concentration of zolpidem in evaluating this strange behaviour. CONCLUSION: Amnesia and incoherence of speech, disorganization of behaviour, inability to realize the situation and mood changes may indicate a zolpidem-induced somnambulism-like state with sleep-related complex behaviour.

Pitfall in cannabinoid analysis--detection of a previously unrecognized interfering compound in human serum.

In clinical and forensic toxicology, high-performance liquid chromatography/tandem mass spectrometry (LC-MS/MS) is increasingly used since it allows the development of sensitive and fast drug analysis procedures. During development of a LC-MS/MS method for determination of the psychoactive cannabinoid Δ(9)-tetrahydrocannabinol (THC) and of its two metabolites 11-hydroxy-THC (THCOH) and 11-nor-9-carboxy-THC (THCCOOH) in serum, a previously unrecognized interfering compound was detected. Extending the fast gradient elution program by an isocratic phase leads to sufficient separation of the interfering compound, initially co-eluting with THCCOOH and exhibiting the same fragments. For characterization, product ion scans and precursor ion scans were performed. Samples from cannabis users were analyzed to estimate the abundance of the interfering compound. The mass spectrometric experiments showed that the interfering compound exhibited the same molecular mass as THCCOOH and a similar fragmentation pattern except for relative fragment intensities. This compound was exclusively detectable in authentic samples. Concentrations were in the range of 4.5 to 51 % (median 14.6 %, n = 73) of those of THCCOOH. After further optimization of the gradient, the method was sufficiently selective and sensitive and validation parameters were within acceptance limits. A new compound related to cannabis use was detected in human serum, and data suggest an isomeric structure to THCCOOH. Considering the rather high amounts observed, it was surprising that this compound had not been detected previously. Further studies on its structure and origin are necessary.

[How relevant is the risk of losing evidence due to delayed blood sampling? Conclusions from the evaluation of blood analysis results]. / Wie relevant ist die Gefahr des Beweismittel- verlustes bei verzögerter Blutentnahme? Schlussfolgerungen aus der Auswertung von Blutuntersuchungsergebnissen.

If the order of a judge to take a blood sample can only be obtained with a marked delay after the incident, evidence proving that a suspect had been driving under the influence of alcohol or drugs of abuse may be lost. The evaluation of blood analysis results from the Institute of Legal Medicine in Frankfurt/Main from the years 2012-2014 shows that in 1.6 to 11.6% of positive cases, the drug concentrations were near the legal limits (20.2% of alcohol-positive and 7.5% of illicit drugs-positive samples). A loss of evidence can thus be expected in a large number of cases when the time between the police check of a driver and the collection of a blood sample increases. Blood concentrations of alcohol and drugs of abuse, especially tetrahydrocannabinol, cocaine, methamphetamine, and morphine, may already have dropped significantly after a delay of only half an hour. These delays are typically due to the time elapsing until the order to take a blood sample has been obtained from a judge and a medical doctor becomes available and arrives at the police station to draw a blood sample. The recommendation of medicolegal experts is to keep the time between police check of a suspect and blood sampling as short as possible. In routine cases, a realistic maximum of one hour should not be exceeded.

Analysis of lysergic acid amide in human serum and urine after ingestion of Argyreia nervosa seeds.

The ergot alkaloid lysergic acid amide (LSA) is a secondary plant constituent in a number of plants, but it is mainly present in considerable amounts in Convolvulaceae, like Argyreia nervosa. Due to its close structural similarity to lysergic acid diethylamide, LSA is considered as psychedelic and therefore promoted as so-called "legal high" in various internet forums. During a human behavioral study with orally administered seeds of A. nervosa, blood and urine samples were obtained. The present study describes the validation of a sensitive and robust high performance liquid chromatography method with fluorescence detection, which was applied to the study samples. The limit of detection (LOD) and lower limit of quantification in human serum were 0.05 and 0.17 ng/mL, respectively, and in urine, the LOD was 0.15 ng/mL. Intra- and interday precision and accuracy were below 15 % relative standard deviation with a bias better than ±15 %. No conversion of LSA to its epimer iso-LSA was noted during analyses. The LSA concentrations in the authentic human serum samples were in the range of 0.66 to 3.15 ng/mL approximately 2 h after ingestion. In urine, LSA could be found 1-24 h after ingestion; after 48 h, no LSA could be detected. The LSA epimer iso-LSA was also detected in serum and urine in varying ratios. In conclusion, LSA serum levels in the low nanogram per milliliter range correlated with severe vegetative adverse effects (nausea, weakness, fatigue, tremor, blood pressure elevation) and a psychosis-like state, which led to study termination.

Plasma and brain levels of terpene trilactones in rats after an oral single dose of standardized Ginkgo biloba extract EGb 761®.

Several studies indicate that the terpene trilactones (TTL) of EGb 761® are responsible for most of its pharmacological action in the brain . Therefore, we investigated the ability of the TTL to cross the blood brain barrier in rats after a single oral administration (600 mg/kg) of EGb 761® and compared it with the plasma levels. In addition, we checked the pharmacokinetic characteristics of an application of EGb 761® against a similar amount of pure substances. For this purpose, we developed a sensitive HPLC-(APCI)-MS method for the determination of the Ginkgo biloba TTL (ginkgolide A [GA], B [GB], C [GC] and bilobalide [Bb]) in plasma as well as in brain tissue. The following animal study shows that the oral application of 600 mg/kg EGb 761® results in significant GA, GB, and Bb concentrations in plasma as well as in the CNS of the rodents, while the GC concentration was below the detection limit of the analytical method in both matrices. GA, GB, and Bb brain concentrations showed a rapid increase up to 55 ng/g, 40 ng/g, and 98 ng/g with no difference of the characteristic after extract or pure substance application. Regarding the plasma levels, significant higher C(max) and AUC values were detected after application of the extract EGb 761®. These results allow for the first time a discussion of pharmacological effects with the knowledge of the pharmacokinetic behavior of the TTL in target tissues.

Assay of ethanol and congener alcohols in serum and beverages by headspace gas chromatography/mass spectrometry.

The analysis of ethanol and of its congeners in blood plays an important role in forensic cases, especially when allegations are made that alcohol has been consumed after an accident. In alcoholic beverages, congener alcohols are by-products and are generated during fermentation. The assay of these compounds in serum samples and beverages has been previously performed using headspace-gas chromatography-flame ionization detection methods (HS-GC-FID). As an alternative, a robust headspace-gas chromatography-mass spectrometry (HS-GC-MS) procedure was developed and validated, which has the following advantages:•Simultaneous determination of ethanol, congener alcohols and other endogenous substances.•Reduction of matrix interference by increasing selectivity and specificity.•Clear separation of the positional isomers 3-methyl-1-butanol and 2-methyl-1-butanol.

Endogenous formation of 1-propanol and methanol after consumption of alcoholic beverages.

INTRODUCTION: In cases of drunk-driving, allegations that alcohol has been consumed after the incident, are proved by analyzing congener alcohols in the blood sample. 1-Propanol, one of the main congener compounds, was tested, whether it is also endogenously formed when a person has consumed alcoholic beverages. METHODS: Eleven male and 13 female volunteers consumed congener-free vodka (37.5 vol% ethanol, individual doses: 0.15-0.32 l) within one hour. Blood samples were taken up to 10 h and analyzed for ethanol and congener alcohols by headspace gas chromatography-mass spectrometry. RESULTS: Ethanol concentrations reached in blood a maximum of 0.65-1.23 g/l and decreased by 0.18 g/l/h (median values). Of the congener alcohols analyzed, only methanol and 1-propanol were detected in the plasma samples of all subjects. The endogenous methanol concentration increased from 0.66 mg/l by 0.22 mg/l/h to 2.19 mg/l (medians). 1-Propanol was not detected prior to alcohol consumption. Maximum concentrations of 0.10-0.32 mg/L were measured after 1.0-4.5 h. A plateau of the 1-propanol concentration was observed in the plasma samples of the 18 subjects lasting for 0.5-4.0 h and this alcohol was completely eliminated at ethanol concentrations of 0.17 g/l (median, range 0.03-0.55 g/l). CONCLUSION: The results of the study confirm the formation of 1-propanol after consumption of 1-propanol-free beverages, which should be taken into account when evaluating its concentration.

Rational design of a pirinixic acid derivative that acts as subtype-selective PPARgamma modulator.

Peroxisome proliferator-activated receptor gamma (PPARgamma) is involved in glucose and lipid homeostasis. PPARgamma agonists are in clinical use for the treatment of type 2 diabetes. Lately, a new class of selective PPARgamma modulators (SPPARgammaMs) was developed, which are believed to show less side effects than full PPARgamma agonists. We have previously shown that alpha-substitution of pirinixic acid, a moderate agonist of PPARalpha and PPARgamma, leads to low micromolar active balanced dual agonists of PPARalpha and PPARgamma. Herein we present modifications of pirinixic acid leading to subtype-selective PPARgamma agonists and furthermore the development of a selective PPARgamma modulator guided by molecular docking studies.

Excretion of 4-fluoroamphetamine and three metabolites in urine after controlled oral ingestion.

Each year, synthetic drugs are occurring in high numbers in the illicit drug market. But data on their pharmacology and toxicology are scarcely available. Therefore, a pilot study was performed to evaluate excretion of 4-fluoroamphetamine (4FA) in humans and identify metabolites in urine. Twelve subjects ingested 100 mg and five 150 mg 4-FA in a bitter lemon drink. Urine samples were scheduled at baseline and 4 times during the following 12 h and analyzed by liquid chromatography-mass spectrometry (LC-MSMS). Concentrations of 4-FA were in the range of 0.7-38 mg/l which is in accordance with the data in previously reported cases. A marked decrease of creatinine excretion in the first two samples was noted. The creatinine normalized concentrations show a maximum 4 h after ingestion in accordance with serum pharmacokinetics. Three products of two metabolic pathways were identified in very low concentrations, two diastereomers of 4-fluorophenylpropanolamine and one ring hydroxylated 4-FA that was conjugated to a large extent. The concentration-time courses paralleled those of 4-FA. The study results show the range of 4-FA concentrations to be expected in urine after oral ingestion of typical dosages and show two pathways of 4-FA metabolism.

l-Thyroxin and the Nonclassical Thyroid Hormone TETRAC Are Potent Activators of PPARγ.

Thyroid hormones (THs) operate numerous physiological processes through modulation of the nuclear thyroid hormone receptors and several other proteins. We report direct activation of the nuclear peroxisome proliferator-activated receptor gamma (PPARγ) and retinoid X receptor (RXR) by classical and nonclassical THs as another molecular activity of THs. The T4 metabolite TETRAC was the most active TH on PPARγ with nanomolar potency and binding affinity. We demonstrate that TETRAC promotes PPARγ/RXR signaling in cell-free, cellular, and in vivo settings. Simultaneous activation of the heterodimer partners PPARγ and RXR resulted in high dimer activation efficacy. Compared to fatty acids as known natural ligands of PPARγ and RXR, TETRAC differs markedly in its molecular structure and the PPARγ-TETRAC complex revealed a distinctive binding mode of the TH. Our observations suggest a potential connection of TH and PPAR signaling through overlapping ligand recognition and may hold implications for TH and PPAR pharmacology.

Chronic alcohol abuse may lead to high skin iron content, but not to hepatic siderosis.

INTRODUCTION: In forensic cases, ante mortem chronic alcohol abuse can be of central importance in clarifying circumstances of death. However, reliable markers of alcohol consumption, which are still available postmortem, are needed. In addition to medical history data which may not be always authentic, the determination of ethyl glucuronide (EtG) in hair as a promising parameter is of no value in cases of missing or cosmetically treated hair. On the other hand, there exist reports that iron ions accumulate in liver tissue (siderosis) during chronic, excessive alcohol consumption, which, therefore, may be useful to serve as alcohol abuse correlate. However, the influence of ethanol on iron stored in the liver has not been adequately investigated and the study situation appears to be inconsistent. AIMS: The aim of the present study was to assess the suitability of assaying iron concentrations in liver and other tissues as postmortem alcoholism marker. METHODS: The iron concentration in tissue samples (liver, brain, skin, pancreas, spleen), vitreous fluid and blood taken during autopsy was analyzed by atomic absorption spectroscopy. The analytical method has been validated before. Cases were divided into two groups: chronic alcohol abusers and non-chronic alcohol consumers including total abstainers using ethyl glucuronide levels in hair as well as anamnestic data as criteria. RESULTS: No elevated iron concentrations in the liver of chronic alcohol abusers were detected. Surprisingly, the iron concentration in skin tissue was found to be significantly higher in cases of chronic alcohol abuse, independent on whether fatty liver or liver cirrhosis was present (as diagnosedduring autopsy). In 18.5% of the cases, chronic alcohol abuse was not confirmed by the EtG concentration in hair. Thus, anamnestic data should not be overestimated. CONCLUSION: The general assumption that chronic alcohol abuse induces hepatic siderosis, i.e. high iron concentrations in liver tissue, has not been supported by results of the present study. However, there seems to exist a correlation between chronic alcohol abuse and high iron concentrations in the skin.

Pharmacokinetic properties of 4-fluoroamphetamine in serum and oral fluid after oral ingestion.

INTRODUCTION: Each year, synthetic drugs occur in high numbers on the illicit drug market. But data on their pharmacology and toxicology are scarce. Therefore, a controlled study was performed to evaluate pharmacokinetic parameters of 4-fluoroamphetamine (4-FA) in humans and to compare it with effects. METHODS: Twelve subjects ingested 100 mg and five subjects also received 150 mg 4-FA in a bitter lemon drink. Blood and oral fluid samples were taken during the following 12 hours and analyzed for 4-FA and traces of amphetamine as impurity by liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: For 12 hours after ingestion, the concentration-time course of 4-FA was similar to that of amphetamine with maximal concentrations appearing in serum after about 2 hours (in median 195 ng/mL after the 100 mg dose, range 155-316 ng/mL). The elimination half-life was approximately 8-9 hours and shorter than that of amphetamine but it exhibited a marked variation (5.5-16.8 hours). In oral fluid, 4-FA could also be detected for 12 hours and concentrations were higher than in serum. During the first 3 hours after ingestion concentrations were higher, most probably due to oral contamination. Serum concentrations in forensic cases were in the range of those observed in the present study suggesting dosages in recreational use in the range of those tested here. CONCLUSIONS: The pharmacokinetic properties of 4-FA are similar to that of amphetamine including a marked variation in elimination. However, recreational dosages may already exhibit prominent adverse effects and may even have life-threatening consequences.

Excretion of metabolites of the synthetic cannabinoid JWH-018 in urine after controlled inhalation.

Each year, synthetic cannabinoids are occurring in high numbers on the illicit drug market but data obtained after controlled application are rare. The present study on pharmacokinetics in urine is part of a pilot study on adverse effects of JWH-018, which is one of the oldest and best known synthetic cannabinoids. Six subjects inhaled smoke from 2 and 3mg JWH-018. The drug and ten potential metabolites were analyzed in urine samples collected during 12h after inhalation by liquid chromatography-mass spectrometry (LC-MS/MS) without and with conjugate cleavage. The parent compound was not detectable, but 13 of its metabolites, all of which were conjugated. Concentrations of the predominant metabolite, JWH-018 pentanoic acid, were less than 5ng/ml, but in two subjects it was still detected up to 4 weeks after ingestion. Other major metabolites were 5- and 4-HOpentyl-JWH-018, JWH-073 butanoic acid and a hypothetically dihydroxylated and dehydrogenated metabolite of JWH-018. Occasionally, further hydroxylated metabolites were found. Generally, hydroxylated metabolites were detected in concentrations lower than 1ng/ml already 10h after inhalation. All concentrations were much lower than reported for urine samples of authentic JWH-018 users. The formation of the metabolite JWH-018 pentanoic acid was found to be slightly delayed, but its rather high concentrations and detection over several weeks after single dosing makes it a useful target for urine analysis. The different excretion of carboxylic acid and hydroxylated metabolites may aid in evaluation of time of use.

Pharmacokinetic properties of the synthetic cannabinoid JWH-018 in oral fluid after inhalation.

Each year, synthetic cannabinoids occur in high numbers on the illicit drug market, but data on their detectability are rarely available. A pilot study was performed to assess adverse effects of JWH-018, which is one of the oldest and best known synthetic cannabinoids. Oral fluid has been evaluated as a specimen for drug monitoring. Six subjects inhaled smoke derived from 2 and 3 mg JWH-018. The drug and 10 of its metabolites were analyzed in oral fluid samples collected during the following 12 hours using the Quantisal collection device by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Maximum concentrations of JWH-018 reached 2.2-2036 (median 25.7) ng/mL after inhalation and decreased during the next hour to only 0.08-8.42 (median 0.89) ng/mL. Metabolites were not found. During the elimination phase (median half-life 1.69 hours), detection of the drug over 6-12 hours (median 8 hours) after inhalation was achieved (0.024 ng/mL limit of quantification). Oral fluid/serum ratios varied considerably intra- and inter-individually in a range of 0.05-555 (median 1.38). The detection of JWH-018 in oral fluid requires high analytical sensitivity even 1 hour after inhalation. The pharmacokinetic properties of inhaled JWH-018 are similar to those of THC. Times for detection are typically less than 12 hours. High variability of the oral fluid/serum ratio precludes extrapolation of oral fluid concentrations to blood.

Influence of bleaching and coloring on ethyl glucuronide content in human hair.

Ethyl glucuronide (EtG) is increasingly used in forensic toxicology as a marker for alcohol use in analyses of hair samples, especially in abstinence control. Some cosmetic treatments are considered to markedly reduce the EtG content. In view of especially many women with coloured hair the present study was performed to further investigate the effect of a variety of colouring procedures (bleaching, tinting, permanent and semi-permanent dyeing, henna) on the EtG content. Untreated hair samples (n = 12, EtG 13.9-64.7 pg/mg) were re-analyzed (gas chromatography- negative chemical ionization mass spectrometry, 0.8 pg/mg quantification limit) after different treatment procedures. A decrease of the EtG content of at least 10% occurred in every case. The reduction in comparison to the untreated hair was expectedly high for permanent dyeing and bleaching with 18.1% of the initial content (median, range 0.0-50.9%) and 18.4% (0.0-46.7%), respectively. For henna this was 38.3% (0.0-83.0%), for tinting 70.4% (29.0-90.8%), for semi-permanent dyeing 41.9% (0.0-77.4%). With permanent hair dye the EtG content was decreased to below 7 pg/mg in 10 of 12 cases, in 3 cases even below the LOD (0.2 pg/mg). Surprisingly henna treatment without oxidative component had a marked influence, EtG was below 2 pg/mg in 2 of 12 samples. The study showed that all tested coloration procedures markedly affected the deposited EtG content. Even temporary or henna coloration may have a marked effect. The present data support the recommendation to exclude hair samples with colour manipulations for analysis on the EtG content as a precaution in alcohol abstinence programs. Copyright © 2017 John Wiley & Sons, Ltd.

11-nor-9-carboxy-Δ9-tetrahydrocannabinol glucuronide exhibits acyl-migration isomers.

11-nor-Δ9-Tetrahydrocannabinol-9-carboxylic acid glucuronide (THCCOOH-glucuronide) is an 1-ß-O-acyl glucuronide which degrades not only to 11-nor-9-carboxy-Δ9-THC (THCCOOH) but, additionally, to an isomer with a currently unknown structure. The present study was carried out to examine whether acyl glucuronide isomers are formed by acyl migration and if they are involved in formation of this isomer. THCCOOH-glucuronide was incubated in phosphate buffer (pH 7.4, 37°C, 7days) and a variety of glucuronide cleavage procedures were performed. Samples of the incubation mixture and of different biological specimens from cannabis users were analyzed using liquid chromatography-mass spectrometry (LC-MS/MS). A total of six chromatographically separated isomeric acyl glucuronides were detected during incubation of THCCOOH-glucuronide reference substance. In biological specimens of cannabis users, two additional isomers were found. However, the main glucuronide present in human specimens was different from that of a commercially available reference substance. Both, the commercial and the authentic glucuronide were cleaved by ß-glucuronidases, the other formed isomers by alkaline hydrolysis only. Mass spectrometric investigations (i.e. product ion, precursor ion and neutral loss scans) confirmed identity. The THCCOOH isomer was detected in all authentic samples, but not in those after buffer incubation. By analyzing THCCOOH-glucuronide in authentic samples, it has to be taken into account that the authentic glucuronide is different from that of the commercial reference standard. THCCOOH-glucuronide undergoes acyl migration and some isomers occur to minor extents in biological specimens. Acyl migration does not lead to the formation of the THCCOOH isomer.

Pharmacokinetic properties of the synthetic cannabinoid JWH-018 and of its metabolites in serum after inhalation.

Each year, synthetic cannabinoids are occurring in high numbers in the illicit drug market, but data on their pharmacology and toxicology are scarcely available. Therefore, a pilot study was performed to assess adverse effects of JWH-018, which is one of the oldest and best known synthetic cannabinoids. Six subjects inhaled smoke from 2 and 3mg JWH-018. The drug and nine of its metabolites were analyzed in their blood samples taken during the following 12h by liquid chromatography-mass spectrometry (LC-MSMS). The maximum concentration of JWH-018 reached 2.9-9.9ng/ml after inhalation and markedly decreased during the next 1.5h, followed by a multiexponential decline (t1/2 in median 1.3h and 5.7h). The concentration of the pentanoic acid metabolite was slightly higher than that of the 3-, 4- and 5-hydroxypentyl metabolites and of the 6-hydroxyindol metabolite. The data also suggest a multiexponential decline and slow terminal elimination of JWH-018 and all metabolites. The detection of JWH-018 and of its metabolites in serum requires high analytical sensitivity. The pharmacokinetic properties of inhaled JWH-018 are similar to that of THC. A slow terminal elimination of drug and metabolites may lead to accumulation in chronic users.