Aerodynamic Thermal Breakup Droplet Ionization in Mass Spectrometric Drug Analysis.

Aerodynamic thermal breakup droplet ionization (ATBDI) in mass spectrometric drug analysis is considered. Cocaine, heroin, and the main alkaloids of opium (morphine, codeine, papaverine) were chosen as the test compounds. The principles of ATBDI ionization are discussed. The dependences of the intensities of the peaks of the target compounds on temperature during ATBDI ionization are also considered. In some cases, a comparison of ATBDI ionization with electrospray ionization (ESI) was performed. In addition, a comparison of methods is demonstrated by the analysis of confiscated opium that was provided by the local police department. Five major alkaloids are found in opium: morphine, codeine, thebaine, papaverine, and narcotine.

Ion mobility spectrometry as a fast screening tool for synthetic cannabinoids to uncover drug trafficking in jail via herbal mixtures, paper, food, and cosmetics.

The greatest challenge for European drug policies is how to effectively respond to the dynamic and constantly changing market for new psychoactive substances (NPS). Even small modifications in the chemical structure of substances often allow circumventing existing laws. Also in prison, the consumption of NPS is rising and there is growing evidence that NPS are responsible for a large share of drug-related problems. Ion mobility spectrometry (IMS) is the technique of choice for trace analysis of illicit drugs or explosives at security points, for example airports. Currently, databases of the reduced mobility (K0 ) values are limited to classical drugs and should be completed with data of emerging NPS. In this article, K0 values, LODs (0.7-3.6 ng) and drift times of 25 synthetic cannabinoids were evaluated. The data were added to existing databases of IMS which were then applied for fast screening in prison. The detection capability of the portable IMS technique was evaluated by the determination of intra-day (0.089%) and inter-day precision (0.004% to 0.14%), systematic error (0.19%), and separation capability for structurally related NPS. The applicability of the methodology was demonstrated by the successful analysis of 12 different pieces of paper impregnated with synthetic cannabinoids, 7 different cosmetics, and 5 food samples (liquids), spiked with a mixture of narcotic drugs and a synthetic cannabinoid. In addition, 14 herbal mixtures and 36 different casework samples from prisons were analyzed provided by the State Office of Criminal Investigation Rhineland-Palatinate (Germany).

Mitragynine concentrations in two fatalities.

Two cases of fatalities are reported of which the recreational use of Mitragyna speciosa ("kratom") could be confirmed. One of these cases presents with one of the highest postmortem mitragynine concentrations published to date. Our results show that even extremely high mitragynine blood concentrations following the consumption of kratom do not necessarily have to be the direct cause of death in such fatalities as a result of an acute overdose. The two cases are compared with regard to the differences in mitragynine concentrations detected and the role of mitragynine in the death of the subjects. Irrespective of the big differences in mitragynine concentrations in the postmortem blood samples, mitragynine was not the primary cause of death in either of the two cases reported here. Additionally, by rough estimation, a significant difference in ratio of mitragynine to its diastereomers in the blood and urine samples between the two cases could be seen.

Suspected Adulteration of Commercial Kratom Products with 7-Hydroxymitragynine.

INTRODUCTION: Kratom (Mitragyna speciosa), a plant native to Southeast Asia, has been used for centuries for its stimulant and opium-like effects. Mitragynine and 7-hydroxymitragynine, exclusive to M. speciosa, are the alkaloids primary responsible for Kratom's biologic and psychoactive profile, and likely contribute to its problematic use. We purchased several commercially available Kratom analogs for analysis and through our results, present evidence of probable adulteration with the highly potent and addictive plant alkaloid, 7-hydroxymitragynine. METHODS: A simple and sensitive LC-MS/MS method was developed for simultaneous quantification of mitragynine and 7-hydroxymitragynine in methanol extract of marketed Kratom supplements. RESULTS: We found multiple commercial Kratom products to have concentrations of 7-hydroxymitragynine that are substantially higher than those found in raw M. speciosa leaves. CONCLUSIONS: We have found multiple packaged commercial Kratom products likely to contain artificially elevated concentrations of 7-hydroxymitragynine, the alkaloid responsible for M. speciosa's concerning mechanistic and side effect profile. This study describes a unique form of product adulteration, which stresses the importance of increased dietary supplement oversight of Kratom-containing supplements.

Rapid identification of a narcotic plant Papaver bracteatum using flow cytometry.

In May 2011, numerous poppy plants closely resembling Papaver bracteatum Lindl., a type of narcotic plant that is illegal in Japan, were distributed directly from several large flower shops or through online shopping throughout Japan, including the Tokyo Metropolitan area. In order to better identify the narcotic plants, the relative nuclear DNA content at the vegetative stage was measured by flow cytometric (FCM) analysis in 3 closely-related species of the genus Papaver section Oxytona, namely P. orientale, P. pseudo-orientale, and P. bracteatum, based on the difference between the chromosome numbers of these species. The results showed that the nuclear DNA content differed between these 3 species, and that most of the commercially distributed plants examined in this study could be identified as P. bracteatum. The remaining plants were P. pseudo-orientale, a non-narcotic plant. In addition, the FCM results for the identification of P. bracteatum completely agreed with the results obtained by the morphological analysis, the inter-genic spacer sequence of rpl16-rpl14 (PS-ID sequence) of chloroplast DNA, and the presence of thebaine. These results clearly indicate the usefulness of FCM analysis for the identification of P. bracteatum plants, including when they are in their vegetative stage.

The analysis of plant-based raw materials of unknown origin.

Chosen aspects of the safety of use of several herbs received from National Medicines Institute, which came from smuggling, have been examined. The analysis has been conducted in three different aspects: (1) Possibilities of contamination of plant-based raw materials by metals of heavy elements (As, Cd, Cu, Cr, Pb). (2) Conscious smuggling of intoxicating preparation or narcotics in plant-based raw materials. (3) Radioactive contamination originating mostly from 137Cs isotope. To solve the problem, analytical methods of GFAAS and ICP-MS, X-ray diffraction and high-distributive spectrometry of gamma-radiation have been applied. Determined concentration of arsenic in all analyzed samples and the concentration of lead in one sample exceeded allowable concentration recommended by WHO. In analyzed materials, no presence of narcotics or radioactive contamination of 137Cs isotope has been detected.

Thebaine in hair as a marker for chronic use of illegal opium poppy substances.

Opium poppy products are often illegally used for both recreational and medicinal purposes. In order to demonstrate the ingestion of opium poppy substances, morphine, codeine and their metabolites have been identified. However, morphine and codeine also originate from the ingestion of therapeutic drugs. Therefore, thebaine, one of the main opium alkaloids, in hair was suggested as a marker for chronic use of illegal opium poppy substances in the present study. First, thebaine was included in the analyte list of our routine analytical method for the simultaneous quantification of codeine, morphine, norcodeine, normorphine and 6-acetylmorphine (6-AM) in hair, which was fully validated previously. Then, the incorporation of thebaine and other opiates into hair and the effect of hair pigmentation were examined using lean Zucker rats with both dark grey and white hair on the same body. Thebaine was also measured in hair samples from actual cases of opium poppy substance use. Consequently, thebaine in hair was demonstrated as a marker of chronic use of illegal opium poppy substances using an animal study and actual cases. Thebaine and other opiates were successfully measured in pigmented hair from rats that ingested raw opium suspension. Moreover, thebaine identified in hair excluded possibility of ingestion of pharmaceutical opiates in actual cases.

HAIRVEQ: an external quality control scheme for drugs of abuse analysis in hair.

The Istituto Superiore di Sanità of Rome, Italy, in cooperation with Institut Municipal d'Investigaciò Mèdica of Barcelona, Spain, set up an external quality control program (HAIRVEQ) to evaluate reliability in hair testing for drug abuse by laboratories from the Italian National Health Service. Samples included in the program were real hair samples from drugs consumers. Prior to sending, hair samples were reduced to powdered form, mixed to ensure homogeneity and tested with GC/MS by four Reference Laboratories. Up to now, four different exercises have been concluded and 23 laboratories participated. Samples containing high and low concentrations of opiates, cocaine and metabolites, low concentrations of MDMA and two blank samples, were included in the intercomparison exercises performed in the first year of HAIRVEQ activities. Results show an insufficient performance of participating laboratories. About 82% of laboratories reported incorrect results on a qualitative basis (false positive and false negative results) for some of the submitted samples. More than one-half of laboratories reported quantitative results (60%). On the basis of the calculated z scores, only between 35 and 55% of results reported should be considered as satisfying. Guidelines have to be provided by Italian authorities for method validation as well as set of recommended cut-off concentrations to orientate laboratories in their quality objectives when developing analytical methodologies as tools to improve reliability and consequently performance of hair analysis.

Simultaneous determination of in total 17 opium alkaloids and opioids in blood and urine by fast liquid chromatography-diode-array detection-fluorescence detection, after solid-phase extraction.

A fast liquid chromatographic method with tandem diode array-fluorescence detection for the simultaneous determination of in total 17 opium alkaloids and opioids is presented. Blank blood and urine samples (1 ml) were spiked with different concentrations of a standard mixture, as well as with the internal standard, butorphanol (2000 ng/ml). After solid-phase extraction, based on weak cation exchange (Bond Elut CBA SPE columns), the extracts were examined by HPLC-DAD-FL. By using a "high-speed" phenyl column (53 x 7.0 mm I.D., particle size 3 microm) eluted with a gradient system (A: water-methanol (90:10, v/v), B: methanol, both containing 25 mM triethylammoniumformate (pH(A) = 4.5)) all compounds could be baseline separated within 12 min. The method was validated and its applicability was demonstrated by the analysis of real-time forensic cases.

Trastornos adictivos y otros trastornos mentales / Addictive and other mental disorders

Diferentes estudios epidemiológicos y clínicos han demostrado una elevada comorbilidad entre los trastornos relacionados con el abuso de drogas y otros trastornos psiquiátricos, como los trastornos de ansiedad, los trastornos del estado de ánimo, los trastornos de personalidad y los trastornos psicóticos. Los pacientes que demandan tratamiento en los centros específicos de drogodependencias se presentan desestructurados cognitivamente, muestran respuestas elevadas de ansiedad y diversos problemas de interacción social y familiar. En muchos casos esta situación se prolonga una vez iniciado el tratamiento y conseguida la abstinencia a drogas. Estos pacientes presentan diversos indicadores de psicopatología que ensombrecen el propósito de abstinencia a largo plazo, son más inestables, presentan un mayor índice de recaídas y realizan numerosos intentos de deshabituación. A continuación se realiza una revisión de distintos trabajos que han encontrado una asociación entre psicopatología y abuso de drogas. Nos centraremos en estudios sobre las sustancias de abuso que generan una mayor demanda de tratamiento: alcohol, heroína, cocaína y anfetaminas. El conocimiento por parte de los clínicos de la incidencia de psicopatología en este tipo de pacientes resulta crucial para planificar sus intervenciones (AU)

Opiate concentrations following the ingestion of poppy seed products--evidence for 'the poppy seed defence'.

The universally accepted 300 ng/ml cut-off limit for opiate assays stated to be mandatory for all drug screening laboratories by the Substance Abuse and Mental Health Services Administration, has been questioned recently due to positive results being obtained following the ingestion of poppy seed containing food products. To establish the plausibility of the 'the poppy seed defence' the concentrations of codeine, norcodeine, morphine, normorphine and thebaine (a potential marker for seed ingestion) in several varieties of poppy seeds from different countries were quantified by GC-MS. The country of origin of the seed specimen analysed and the preparation of the seeds prior to their culinary use was found to influence the alkaloid concentration determined. The maximum morphine and codeine concentrations determined in the seeds were found to be 33.2 and 13.7 micrograms/g seed respectively. In addition, thebaine concentrations were found to vary with each seed sample analysed. Following the consumption of bread rolls (mean 0.76 g seed covering per roll) by four subjects, all urine specimens analysed produced negative results (using the Dade Bebring EMIT II opiate screening assay) with the exception of one subject (body weight 63.0 kg) who consumed two poppy seed rolls. In this subject opiate positive screening results were obtained for up to 6 h post ingestion with maximum urinary morphine and codeine concentrations of 832.0 ng/ml (@ 2-4 h post ingestion) and 47.9 ng/ml (@ 0-2 h post ingestion) respectively being achieved. Following the ingestion of poppy seed cake containing an average of 4.69 g of seed per slice by four individuals, opiate positive screening results were obtained for up to 24 h. In one subject (dose equivalent to 0.07 g poppy seed/kg body weight) maximum urinary morphine and codeine concentrations of 302.1 ng/ml (@ 0-2 h) and 83.8 ng/ml (@ 2-4 h) respectively were recorded. The elimination of thebaine was found to vary widely between individuals, therefore suggesting that its absence from a specimen is not necessarily indicative of opiate abuse. These findings demonstrate that the poppy seed defence could be used as an argument in medico-legal and employment medical cases. Great care should therefore be taken when interpreting the data produced when screening for opiates.

Transcranial electrostimulation effects on rat opioid and neurotransmitter levels.

A specific form of Transcranial Electrostimulation Treatment (TCET) has been shown to induce analgesia, alleviate symptoms of opiate withdrawal and alter nociceptive responses in neurons in the midbrain and hypothalamus of rats. TCET consists of a 10Hz, charge balanced, 10 mu A current passed for 30 minutes between electrodes placed in the ears. Both serotonin (5HT) and endogenous opioids have been strongly implicated in TCET responses. This study directly measured brain levels of several neurotransmitters and their metabolites in anesthetized rats stimulated with either 10 mu A TCET or 0 mu A (Sham). Neurotransmitters measured in selected homogenized brain areas by high performance liquid chromatography were 5HT and its metabolite, 5-hydroxyindolacetic acid (5HIAA); norepinephrine (NE) and its metabolite, 3-methoxy-4-hydroxyphenethyleneglycol (MHPG); and dopamine (DA). Levels of NE and DA were significantly higher in the hypothalamic region of TCET rats than of control rats. The midbrains of TCET rats contained significantly elevated levels of DA, MHPG, 5HT and 5HIAA. In the hindbrain no significant differences were observed. Thus, TCET appears to cause an increase in the synthesis or release of 5HT, DA and NE in the midbrain and DA and 5HT in the hypothalamus. In a separate experiment, beta-endorphin-like immunoreactivity was measured in blood plasma taken from rats at intervals before, during and after a 30 minute TCET treatment, but no demonstrable TCET effect was observed. The lack of change in serum endorphin levels suggests that TCET-induced opioid activity may be confined to the central nervous system, a reasonable theory because the current passes only through the head.

An evaluation of TLC systems for opiate analysis.

The authors selected 38 thin-layer-chromatography (TLC) systems described in the available literature published over the last 10 years and evaluated those systems with respect to their suitability for detection and identification of opiates in urine, opium and heroin, as well as adulterants in heroin. A total of 14 substances: 8 opiates (morphine, 6-monoacetylmorphine, diacetylmorphine, codeine, acetylcodeine, noscapine, papaverine and thebaine) and 6 adulterants (ephedrine, quinine, methadone, caffeine, cocaine and strychnine) were used as test samples for this research. Using laboratory-coated plates and pre-coated plates, 15 and 13 TLC systems, respectively, were found to be able to detect and identify morphine and codeine in urine without interference from the remaining 12 substances. For the detection of opiates in opium samples as well as opiates and adulterants in illicit heroin samples the TLC system: chloroform-n-hexane-triethylamine (9:9:4) which was developed by the National Drug Research Centre, Penang, Malaysia, was found to be most suitable on both laboratory-coated and pre-coated plates. In addition, the following two systems, one on laboratory-coated plates--hexane-chloroform-diethylamine (50:30:7)--and the other on pre-coated plates--benzene-dioxane-ethanol-ammonia (50:40:5:5; T-7)--were also found to be among most suitable TLC systems for the analysis of opiates in opium samples. The article also presents the relative cost of each of the 38 evaluated TLC systems.