Gamma-hydroxybutyrate (GHB) is an endogenous compound in the human body, found in regions of the mammalian brain and believed to be a cerebral neurotransmitter.1,2 GHB also acts as a powerful central nervous system depressant commonly used as a “date rape” drug due to its hypnotic and sedative properties.1 The drug has also been used medicinally to treat alcohol withdrawal, opiate-withdrawal syndrome, and narcolepsy.3–5 Toxicological analysis of GHB in drug facilitated sexual assault (DFSA) cases is typically performed using blood and urine specimens.1 However, due to the endogenous nature of GHB, toxicological interpretation of these biological specimens can be complex and challenging.1,4 Additionally, urine and blood analysis of GHB can be impacted by sample collection, sample analysis times, and sample storage conditions.6,7 Due to the challenges and limitations associated with blood and urine analysis of GHB along with the prominence of GHB in DFSA cases, it would be beneficial to determine the possibility of GHB analysis using alternative biological matrices. The primary goal of this research was to develop a sample preparation method that could accurately and reliably identify and quantify GHB in oral fluid, as an alternative biological matrix. Additionally, this research was carried out to compare the identification and quantitation capabilities of GHB in oral fluid to that of traditional biological matrices, specifically urine and blood. The methods employed in this study utilized gas chromatography – mass spectrometry (GC-MS) instrumentation in order to correctly identify GHB. A deuterated internal standard, GHB-d6, was used to quantify all samples. The methods were assessed using the parameters set forth by the Scientific Working Group of Forensic Toxicologists (SWGTOX) for quantitative analysis methods. The following factors were considered: calibration model, bias, precision, limit of detection and quantitation, carryover, and interferences. Urine and blood samples were prepared using 200 uL of urine (UTAK Laboratories, Inc., Valencia, CA, U.S.A.) or blood (Equitech-Bio Inc., Kerrville, TX, U.S.A.), varying amounts of the 200 mg/L working calibrator and control solution prepared using certified reference standards (Cerilliant, Round Rock, TX, U.S.A.), and 50 uL of 100 mg/L working internal standard solution resulting in an internal standard concentration of 25 mg/L in each sample. Solid phase extraction (SPE) was performed using United Chemical Technologies (UCT), Inc. (Bristol, PA, U.S.A) Clean Screen GHB columns (ZSGHB020) on all samples.1 Samples were reconstituted, derivatized, and analyzed using GC-MS. Oral fluid samples were prepared using 1.0 uL of drug-free oral fluid, and 1.0 mg/mL (as salt) in methanol GHB received from Cerilliant. The samples were spiked with 1 uL of 1.0 mg/mL (as salt) in methanol GHB-d6 received from Cerilliant. Each sample had an internal standard concentration of 10 mg/L. Samples were fortified with 100 uL of ethyl acetate, and derivatized with 100 uL of bis(trimethyl)trifluoroacetamide (BSTFA) with 1% Trimethylchlorosilane (TMCS) received from Cerilliant. The samples were incubated, and analyzed using GC-MS.8 All analyses were conducted using an Agilent 7890A GC system, Agilent 5975C Mass Detector System (MSD), and an Agilent 7683B Autosampler (Agilent Technologies Inc. Santa Clara, CA). The chromatographic component was carried out using an Agilent HP5-MS 30m x 250um x 0.25um capillary column and an Agilent HP- 5MS 15m x 250um x 0.25um capillary column. All data was analyzed using Agilent MSD ChemStation software (version E.02.02.1431). The method has a total length of 12.75 minutes. Selective ion monitoring (SIM) was used to monitor the ions of interest for each analyte. GHB-d6 was monitored using the ions 239, 240, and 241. GHB was monitored using the ions 233, 234, 235.1 Results revealed that GHB and GHB-d6 could be identified and differentiated due to their fragmentation patterns. All calibration curves for the three matrices exhibited R2 values > 0.98 using a linear dynamic range of 5-100 mg/L with a minimum of four calibration points. The limit of detection for the three matrices was determined to be 1 mg/L, and the limit of quantitation for the three matrices was determined to be 5 mg/L. Bias and precision were analyzed at concentrations of 8 mg/L, 45 mg/L, and 90 mg/L for each matrix. All urine and blood samples were calculated to be within the acceptance range of +20% bias and +20% coefficient of variation. Oral fluid samples were outside of the +20% acceptance range for both bias and coefficient of variation. The highest concentration analyzed that did not produce carryover into subsequent matrix blanks was found to be 350 mg/L for each matrix. Significant interferences were found to be present in urine and blood samples, but negligible for all oral fluid samples. This research illustrates that the developed sample preparation method can be used to accurately and reliably identify GHB in oral fluid. Additionally, this research suggests that the quantitation capabilities of GHB in oral fluid are not as accurate and precise as those of urine and blood. Therefore, the developed method has better qualitative analysis capabilities, while the urine and blood methods have better quantitative analysis capabilities for forensic toxicology casework.

A pesar del uso terapéutico de GHB y de un aumento en la percepción de su toxicidad, el conocimiento sobre los efectos fisiológicos, subjetivos y sobre el rendimiento psicomotor inducidos por el GHB en humanos era limitado. Si bien se habían descrito casos de abuso y dependencia, no se habían realizado estudios controlados evaluando el potencial de abuso de la sustancia. El objetivo más importante de este trabajo fue la caracterización de los efectos del GHB en humanos. En este contexto se pusieron en marcha una serie de estudios controlados para evaluar los efectos del GHB y su potencial de abuso. Los estudios fueron randomizados, a doble ciego, de tipo cruzado y controlados. Las variables estudiadas incluyeron constantes vitales (presión arterial, frecuencia cardiaca, temperatura oral, diámetro pupilar), efectos sobre el rendimiento psicomotor (test de sustitución de símbolos por dígitos, tarea de balance, ala de maddox) efectos subjetivos (cuestionario Addiction Research Center Inventory reducido de 49 items, 13 escalas visuales analógicas, cuestionario de valoración de efectos subjetivos de sustancias con potencial de abuso‐VESSPA) y evaluación farmacocinética. En una primera fase se realizó un estudio piloto de farmacología humana de fase I, donde fueron administradas diferentes dosis de GHB sódico (40, 50, 60 y72 mg/kg) por vía oral a 8 voluntarios en una pauta de dosis ascendente. Las concentraciones de GHB en plasma, fluido oral, orina y sudor fueron analizadas mediante cromatografía de gases acoplada a espectrometría de masas. El método de detección de GHB en plasma, fluido oral y orina fue validado para su posterior uso durante los estudios. Los efectos fisiológicos, subjetivos y sobre el rendimiento psicomotor del GHB fueron evaluados simultáneamente. El GHB produjo efectos de tipo mixto estimulante y sedante, con un incremento inicial de las puntuaciones en los efectos subjetivos de euforia y"colocón", seguido de efectos moderados de tipo sedante asociados a una alteración del rendimiento psicomotor. La administración de GHB se siguió de una rápida absorción y eliminación.El estudio definitivo se realizó con la intención de confirmar los efectos fisiológicos y subjetivos del GHB y su impacto sobre el rendimiento psicomotor, así como pare evaluar su potencial de abuso en comparación con etanol y flunitrazepam, en usuarios de "Club Drugs". Con estos objetivos se evaluaron 12 voluntarios sanos con experiencia previa en el uso de GHB durante 5 sesiones experimentales. Las diferentes condiciones de tratamiento fueron, 2 dosis únicas de GHB (40 o 60 mg/kg), etanol (0.7 g/kg), flunitrazepam (1.25 mg), y placebo, todos administrados por vía oral.Todos los tratamientos activos indujeron efectos de tipo positivo relacionados con su potencial de abuso. La administración de GHB indujo efectos de tipo euforizantes y placenteros ligeramente superiores a los observados tras la administración de flunitrazepam y etanol. El perfil de efectos inducidos por el GHB fue de tipo bifásico, inicialmente de tipo estimulante‐euforizante en relación con el incremento simultáneo de las concentraciones plasmáticas, seguido de un efecto de tipo sedante no relacionado con la cinética plasmática. La administración de GHB produjo asimismo efectos adversos dosis dependientes, pero sin una coincidencia intra‐sujeto de efectos positivos y negativos. Las concentraciones plasmáticas de GHB y las concentraciones de etanol en sangre se correlacionaron significativamente con los efectos subjetivos estimulantes, mientras que las concentraciones plasmáticas de flunitrazepam se correlacionaron significativamente con efectos de tipo sedante. Las concentraciones plasmáticas de GHB se correlacionaron significativamente con las variables relacionadas con el potencial de abuso. En cuanto a los efectos fisiológicos, el GHB indujo un aumento significativo de la presión arterial y del diámetro pupilar, mientras el etanol indujo sus efectos prototípicos y flunitrazepam produjo una marcada sedación. GHB y flunitrazepam produjeron un empeoramiento significativo del rendimiento psicomotor (tareas de sustitución de símbolos por dígitos y del balance), mientras que el etanol únicamente indujo un leve empeoramiento de la tarea del balance.Como conclusión, a las dosis investigadas, la administración de GHB indujo efectos de tipo euforizantes y placenteros, sedación y efectos estimulantes de tipo moderado, similares a los descritos previamente por los usuarios de las sustancia. El perfil de efectos inducidos por el GHB fue de tipo bifásico, inicialmente de tipo estimulante‐eufórico y relacionado con el incremento simultáneo de las concentraciones plasmáticas, seguido de un efecto de tipo sedante no relacionado con la cinética plasmática. La tolerabilidad del GHB difirió substancialmente entre los diferentes sujetos, sin coincidencia intra‐sujeto de efectos positivos y negativos. La administración de GHB se sigue de una rápida absorción y eliminación con una gran variabilidad inter‐indiviudal. Si bien se encontraron concentraciones de GHB todas las matrices biológicas analizadas, tanto el fluido oral como el sudor no parecen convenientes para monitorizar el consumo de GHB. Los resultados sugieren un alto potencial de abuso de GHB en usuarios de "Club Drugs" y aportan los fundamentos científicos del aumento en el abuso de la sustancia en humanos. / Despite GHB therapeutic uses and the increasing concern about the toxicity of this drug, little was known about the physiologic and subjective effects and alterations in psychomotor performance induced by GHB in humans. Furthermore, a number of cases of GHB abuse and dependence have been reported, but no abuse liability studies had been performed. The main aim of this work was characterising the actual behavioural effects of GHB in humans. Within this context, a series of controlled studies to evaluate the effects of GHB and its relative abuse liability were set up. The studies were double blind, randomized, crossover and controlled. Study assessments included vital signs (BP, HR, oral temperature, pupil diameter), psychomotor performance (digit-symbol-substitution-test, balance, maddox‐wing), subjective effects (a set of 13 visual analogue scales, Addiction Research Center Inventory‐49 items, and Evaluation of the Subjective Effects of Substances with Potential of Abuse questionnaires), and pharmacokinetics. Firstly, a pilot pharmacology phase I study was performed where different oral doses of sodium GHB (40, 50, 60, and 72 mg/kg) were given to 8 volunteers in a dose escalation schedule. GHB concentrations in plasma, oral fluid, urine, and sweat were analyzed by gas chromatography-mass spectrometry. The method of detection of GHB in plasma, urine and oral fluid used throughout the studies was validated. Physiological effects, psychomotor performance, and subjective effects were examined simultaneously. GHB showed a mixed stimulant‐sedative pattern, with initially increased scores in subjective feeling of euphoria, high, and liking followed by mild‐moderate symptoms of sedation with impairment of psychomotor performance. GHB was readily absorbed after oral administration and rapidly eliminated. The final study was performed to confirm GHB‐induced subjective and physiological effects, and to evaluate its relative abuse liability compared to flunitrazepam and ethanol, and its impact on psychomotor performance in 'Club Drug' users. Twelve healthy male recreational users of GHB participated in 5 experimental sessions. Drug conditions were a single oral dose of GHB (40 or 60 mg/kg), ethanol (0.7 g/kg), flunitrazepam (1.25 mg), and placebo. All active conditions induced positive effects related to their abuse potential. The administration of GHB produced euphoria and pleasurable effects with slightly higher ratings than those observed for flunitrazepam and ethanol. GHB induced a biphasic time profile with an initial stimulant‐like effect related to the simultaneous rise of plasma concentrations and a latter sedative effect not related to GHB kinetics. GHB administration also induced dose‐dependent mild unpleasant effects, with no within‐subject coincidence of positive and negative GHB related effects. GHB plasma concentrations and ethanol blood concentrations were well correlated to subjective effects related with stimulation whereas flunitrazepam plasma concentrations were better correlated to sedative‐like effects. GHB plasma concentrations were also well correlated to different VAS related to abuse potential. GHB increased blood pressure and pupil diameter. Ethanol induced its prototypical effects, and flunitrazepam produced marked sedation. GHB and flunitrazepam impaired psychomotor performance (digit symbol substitution test and balance task), whereas ethanol, at the dose tested, induced only mild effects exclusively affecting the balance task.In conclusion, at the doses tested GHB was capable of inducing euphoria, pleasurable feelings, sedation and slight stimulant‐like effects as previously reported by GHB users. GHB induced a biphasic time profile with an initial stimulant‐like, euphoric and pleasurable effect related to the simultaneous rise of plasma drug concentrations and ulterior sedative effect collateral to a decrease in GHB plasma concentrations. GHB tolerability highly differ between subjects with no within‐subject coincidence of positive and negative GHB related effects. Following oral administration, GHB is rapidly absorbed and eliminated with high interindiviidual variability. Measurable plasma, urine, oral fluid and sweat were observed. However, oral fluid and sweat appear not to be suitable biological matrices for monitoring GHB consumption. The results suggest a high abuse liability of GHB in 'Club Drug' users, providing the scientific rationale for the increased abuse of the drug in humans.

Gamma Hydroxybutyrate (GHB) was banned from the consumer market by the Food and Drug Administration in 1991. Despite the ban, use of GHB has continued to contribute to thousands of emergency department visits and numerous fatalities in recent years. Efforts to reduce the use of this drug have had limited impact, which may be the result of using traditional prevention strategies that focus exclusively on educating people about of negative consequences of substance use rather than addressing the factors that motivate use. In an effort to identify motivational factors that could be targeted in future prevention efforts, the present study was designed to examine outcome expectancies for GHB that may promote use of this drug. Methodology that has led to successful strategies to reduce alcohol use was applied to identify GHB expectancies and model cognitive processes likely to encourage or discourage GHB use. Individual differences scaling was used to empirically model a two dimensional semantic network of GHB expectancies stored in memory, and preference mapping was used to model likely paths of expectancy activation for male and female GHB users and nonusers. Differences in expectancies between GHB users and nonusers followed patterns previously identified in relation to alcohol expectancies and alcohol use. Conclusions were limited by relatively low numbers of GHB users in the sample, despite the use of a very large number of participants, overall. Despite this limitation these findings lay the groundwork for development and validation of GHB expectancy based prevention strategies.

Cette thèse montre que le GHB et les neurostéroïdes protègent efficacement contre la mort neuronale induite par les facteurs étiologiques de la maladie d'Alzheimer, notamment la sur-expression de l'amyloid precursor protein et le stress oxydant. Nous avons identifié un effet additif du GHB et de l'alloprégnanolone qui pourrait résulter de la combinaison des stimulations partielles évoquées par ces molécules sur l'expression des protéines anti-apoptotiques. L'effet du GHB est bloqué par un inhibiteur de l'aromatase, suggérant que le GHB induirait la neuroprotection via la stimulation de la neurostéroïdogenèse. Pour étudier les effets du GHB et des neurostéroïdes sur l’activité des MMP-2 et MMP-9, qui dégradent les peptides amyloïdes, nous avons optimisé un test enzymatique basé sur l’expression de ces protéases dans la levure. Nos résultats préliminaires ne permettent pas encore de conclure mais leur amélioration et combinaison avec des données de RT-qPCR contribueront à déterminer l'action du GHB et des neurostéroïdes sur l'activité et/ou l'expression des MMP. Notre travail suggère que le GHB et les neurostéroïdes pourraient être associés pour élaborer des stratégies neuroprotectrices contre les pertes neuronales provoquées par la maladie d'Alzheimer. / This PhD work showed that GHB and neurosteroids efficiently protect against nerve cell death caused by Alzheimer's disease etiological factors including amyloid precursor protein overexpression and oxidative stress. Interestingly, we identified an additive action of GHB and allopregnanolone that may result from the combination of partial stimulations of anti-apoptotic protein expression induced by both compounds. GHB protective effect was blocked by aromatase inhibitor, suggesting that GHB may also induce neuroprotection via the activation of neurosteroidogenesis. Finally, we have used a yeast-based MMP activity assay to check whether GHB and neurosteroids regulate MMP-2 and MMP-9 activities which control Aβ peptide degradation. We cannot yet conclude from our preliminary results but their improvement and combination with RT-qPCR analyzes will help to determine the modulatory action of GHB and neurosteroids on MMP activity and/or expression. Together, our data suggest that GHB and neurosteroids may be used to develop combined neuroprotective strategies against neuronal loss in AD.

Hot och våld är vanligt förekommande inom prehospital akutsjukvård. Intoxikation med droger utgör en allt vanligare patientgrupp som vårdas av ambulanspersonal. Ambulansuppdrag med patienter som är intoxikerade med GHB, GBL eller 1.4 BD är vanligt förekommande i Västra Götaland. Hot och våldssituationer i samband med omhändertagandet av GHB-intoxikerade patienter är riskfyllt för ambulanspersonalen. Patienten kan snabbt pendla mellan att vara medvetslös till att bli hotfull. Syftet med studien är att kartlägga prehospitala vårdsituationer med patienter som är intoxikerade med GHB-preparat avseende hot, våld samt risk för hot och våld. Studien har en kvantitativ ansats med en deskriptiv och explorativ design. Totalt analyserades 50 ambulansdatajournaler. Dessa utgjorde dokumentation av en konsekutiv patientgrupp under tiden januari 2009 till augusti 2009. I resultatet påvisas en hög förekomst av hot och våld i samband med prehospitalt omhändertagande av patienter intoxikerade med GHB-preparat. Dokumentationen uppfyllde kraven enligt patientdatalagen, men omfattande kvalitetsbrister kunde ses. Avvikelserapporteringen för den studerade perioden visade två avvikelser gällande hot och våld i samband med vård av patienter intoxikerade med narkotika. Detta pekar på en kraftig underrapportering av hot och våld. / Program: Specialistsjuksköterskeutbildning med inriktning mot ambulanssjukvård

No description available.

Orientadora: Elizabete Campos de Lima / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Ciência e Tecnologia/ Química

<p>En metod för detektering och kvantifiering av <em>gamma</em>-hydroxysmörsyra (GHB) i urin med gaskromatografi (GC) är framtagen på Sahlgrenska universitetssjukhuset. Metoden är relativt unik då den inte kräver upparbetning i form av derivatisering, indunstning eller extraktion. Urinen surgörs med koncentrerad saltsyra och internstandard, <em>gamma</em>-valerolakton, tillsätts. GHB övergår då till laktonformen, <em>gamma</em>-butyrolakton (GBL). Därefter injiceras provet direkt på en GC-FID med en kapillärkolonn för glykoler och alkoholer. Detektion ner till 100 μmol/L är möjligt med en variationskoefficient mellan 6 och 12 %. Provsvar erhålls efter 6,5 minuter. Metoden är dock inte fullständig då en del frågetecken kvarstår. Bland annat bör det undersökas om andra föreningar, som kan förekomma i urin, kan eluera samtidigt som GHB. Om ja så bör vidare analyser genomföras för att separera GHB och den andra föreningen. Metoden kan däremot användas i nuläget som en screeninganalys för att snabbt få ett svar på om GHB finns närvarande eller inte. Verifiering kan sedan ske med GC-MS.</p> / <p>A method for determination and quantification of <em>gamma</em>-hydroxyburyric acid (GHB) in urine samples is developed at Sahlgrenska universitetssjukhus. No time consuming procedures as derivatization and exctration is required, which makes the method fairly unique. Hydrochloric acid and internal standard, <em>gamma</em>-valerolakton, is added to the urine sample before the sample is injected to a gas chromatograph with a flame ionization detector and a column for glycols and alcohols. The hydrochloric acid makes the GHB convert into <em>gamma</em>-butyrolactone (GBL) which is easier to separate in the gas chromatograph. Limit of detection was found to be 100 μmol/L and test result is received after 6,5 minutes. There are still some question marks around the method, for example, there is a possibility that another substance elute at the same time as GHB. More tests are required to determine whether or not it is so. For now the method can be used as a screening analysis to hastily detect GHB presence. Verification can be done with GC-MS.</p>

The research presented in this thesis started with the idea to study alcohols as modifiers and dopants in differential ion mobility spectrometry (d-IMS) to produce complicated chemical signatures to explore a concept of chemical labels for product security application. D-IMS is a gas phase atmospheric pressure separation and detection technique which distinguishes compounds based on differences in their ions mobility as their travel under a low and high electric field. The hypothesis was that alcohols will form typical d-IMS products such as protonated monomers and proton bound cluster ions. However, the very first experiments revealed unexpected phenomena which included changes in the mobility of ions over a narrow range of concentrations that could not be explained by existing theory. Another observation was the apparent regeneration of reactant ions. It became evident that the observed phenomena had not been described in the open literature and that addressing the research-questions that were being raised would be essential for the determination of alcohols by d-IMS and its use in medical applications for toxicity screening and monitoring of alcohols. The above discovery shifted the research objective towards a fundamental and comprehensive study on the behaviour of alcohols in d-IMS. This thesis describes designed experiments and constructed systems allowing the efficient study of effect of concentration, electric field and temperature on the d-IMS responses of alcohols. The results of those studies demonstate: extensive fragmentation of alcohols, including previously undescribed fragmentation patterns with regeneration of the hydrated proton; new phenomena of adduct ion formation within the d-IMS drift tube, observed in the case of methanol within a narrow range of concentration; and self-modification of the alpha function of alcohols. This knowledge was exploited by developing an non-invasive analytical method for recovery, separation and detection of toxins from human saliva (including alcohols, diols and GHB) using TD-GC-d-IMS (thermal desorption - gas chromatography d-IMS) within a full range of toxicological concentration levels.

Objective: The aim was to summarize the clinical literature on interactions between common illicit drugs and prescription therapies. Methods: Medline, Iowa Drug Information Service, International Pharmaceutical Abstracts, EBSCO Academic Search Premier, and Google Scholar were searched from date of origin of database to March 2011. Search terms were cocaine, marijuana, cannabis, methamphetamine, amphetamine, ecstasy, N-methyl-3,4- methylenedioxymethamphetamine, methylenedioxymethamphetamine, heroin, gamma-hydroxybutyrate, sodium oxybate, and combined with interactions, drug interactions, and drugdrug interactions. This review focuses on established clinical evidence. All applicable full-text English language articles and abstracts found were evaluated and included in the review as appropriate. Results: The interactions of illicit drugs with prescription therapies have the ability to potentiate or attenuate the effects of both the illicit agent and/or the prescription therapeutic agent, which can lead to toxic effects or a reduction in the prescription agent's therapeutic activity. Most texts and databases focus on theoretical or probable interactions due to the kinetic properties of the drugs and do not fully explore the pharmacodynamic and clinical implications of these interactions. Clinical trials with coadministration of illicit drugs and prescription drugs are discussed along with case reports that demonstrate a potential interaction between agents. The illicit drugs discussed are cocaine, marijuana, amphetamines, methylenedioxymethamphetamine, heroin, and sodium oxybate. Conclusion: Although the use of illicit drugs is widespread, there are little experimental or clinical data regarding the effects of these agents on common prescription therapies. Scientific Significance: Potential drug interactions between illicit drugs and prescription drugs are described and evaluated on the Drug Interaction Probability Scale by Horn and Hansten.