Original analytical methods for the determination of psychoactive compounds in complex matrices

This thesis work aims to develop original analytical methods for the determination of drugs with a potential for abuse, for the analysis of substances used in the pharmacological treatment of drug addiction in biological samples and for the monitoring of potentially toxic compounds added to street drugs. In fact reliable analytical techniques can play an important role in this setting. They can be employed to reveal drug intake, allowing the identification of drug users and to assess drug blood levels, assisting physicians in the management of the treatment. Pharmacological therapy needs to be carefully monitored indeed in order to optimize the dose scheduling according to the specific needs of the patient and to discourage improper use of the medication. In particular, different methods have been developed for the detection of gamma-hydroxybutiric acid (GHB), prescribed for the treatment of alcohol addiction, of glucocorticoids, one of the most abused pharmaceutical class to enhance sport performance and of adulterants, pharmacologically active compounds added to illicit drugs for recreational purposes. All the presented methods are based on capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) coupled to various detectors (diode array detector, mass spectrometer). Biological samples pre-treatment was carried out using different extraction techniques, liquid-liquid extraction (LLE) and solid phase extraction (SPE). Different matrices have been considered: human plasma, dried blood spots, human urine, simulated street drugs. These developed analytical methods are individually described and discussed in this thesis work.

Rapid and simple LC-MS/MS-Screening of 64 novel psychoactive substances using dried blood spots

The range of novel psychoactive substances (NPS) including phenethylamines, cathinones, piperazines, tryptamines, etc. is continuously growing. Therefore, fast and reliable screening methods for these compounds are essential and needed. The use of dried blood spots (DBS) for a fast straightforward approach helps to simplify and shorten sample preparation significantly. DBS were produced from 10 µl of whole blood and extracted offline with 500 µl methanol followed by evaporation and reconstitution in mobile phase. Reversed-phase chromatographic separation and mass spectrometric detection (RP-LC-MS/MS) was achieved within a run time of 10 min. The screening method was validated by evaluating the following parameters: limit of detection (LOD), matrix effect, selectivity and specificity, extraction efficiency, and short-term and long-term stability. Furthermore, the method was applied to authentic samples and results were compared with those obtained with a validated whole blood method used for Routine analysis of NPS. LOD was between 1 and 10 ng/ml. No interference from Matrix compounds was observed. The method was proven to be specific and selective for the analytes, although with limitations for 3-FMC/flephedrone and MDDMA/MDEA. Mean extraction efficiency was 84.6 %. All substances were stable in DBS for at least a week when cooled. Cooling was essential for the stability of cathinones. Prepared samples were stable for at least 3 days. Comparison to the validated whole blood method yielded similar results. DBS were shown to be useful in developing a rapid screening method for NPS with simplified sample preparation. Copyright © 2013 John Wiley & Sons, Ltd

Behavioral effects of plasma tryptophan depletion in aggressive and nonaggressive men

Serotonin (5-HT) neurotransmission deficits have been implicated in impulsive aggression. A Trp-free beverage of amino acids competitively inhibits Trp uptake into the brain for 5-HT synthesis and also lowers endogenous plasma Trp for several hours. This has worsened mood and/or increased aggressive behavior, especially in hostile persons or those with histories of depression. In 24 community-recruited men (12 each with and without significant aggression histories), aggressive and impulsive behavior in the laboratory was assessed before and after plasma Trp depletion and Trp loading. In the aggression model, subjects were provoked by periodic subtractions of participation earnings, and these subtractions were blamed on a ficitious other participant. Aggression was measured as the responses the subject made to subtract money from his antagonist. Impulsiveness was operationalized as: (1) the choice of smaller reward after a shorter delay over having to wait longer to receive a larger reward, and (2) “false alarm” commission errors in a modified Continuous Performance Task, which represent a failure to inhibit responding to stimuli similar (but not identical) to target stimuli. Finally, plasma cortisol and Trp were measured under each condition immediately following a aggression testing session when subjects were highly provoked. I hypothesized that 5-HT may tonically modulate (inhibit) the hypothalmnic-pituitary-adrenal stress response, such that Trp depletion may enhance the cortisol response to high provocation in aggressive men. ^ Trp depletion had no effect in the laboratory tasks purported to measure impulsive behavior, and failed to cause increases in aggressive behavior under low provocation conditions. Under higher provocation, however, aggressive responses we re elevated under Trp-depleted conditions relative to Trp-loaded conditions in aggressive men, whereas the reverse was true in nonaggressive men. Cortisol levels nonsignificantly paralled the group differences in aggression under Trp-depleted and Trp-loaded conditions. Aggressive men achieved lower plasma Trp levels after Trp loading than did nonaggressive men, possibly due to heavy alcohol use histories. The high post-loading plasma Trp levels in nonaggressive men tended also to correlate with their aggressive responding rates, due perhaps to increases in other psychoactive Trp metabolites. ^

Detection and quantification of 56 new psychoactive substances in whole blood and urine by LC-MS/MS

BACKGROUND New psychoactive substances (NPS) have become increasingly prevalent and are sold in internet shops as 'bath salts' or 'research chemicals' and comprehensive bioanalytical methods are needed for their detection. METHODOLOGY We developed and validated a method using LC and MS/MS to quantify 56 NPS in blood and urine, including amphetamine derivatives, 2C compounds, aminoindanes, cathinones, piperazines, tryptamines, dissociatives and others. Instrumentation included a Synergi Polar-RP column (Phenomenex) and a 3200 QTrap mass spectrometer (AB Sciex). Run time was 20 min. CONCLUSION A novel method is presented for the unambiguous identification and quantification of 56 NPS in blood and urine samples in clinical and forensic cases, e.g., intoxications or driving under the influence of drugs.

Sugar sweetened beverage consumption and behavioral correlates in a pilot study of high school students in Austin, Texas

Habitual consumption of sugar-sweetened beverages (SSB) has been reliably linked to obesity in adolescents. A wide variety of beverages sweetened with sugar are available to this population. The objective of this secondary data analysis was to assess the consumption of SSB by category and to identify behaviors that occur concurrently with the consumption of soda, sport drinks and fruit-flavored drinks in high school students. The analysis used self-reported survey data from 97 adolescents ages 14 to 18. SSB categories considered in the consumption analysis included regular soda, sports drinks, fruit-flavored drinks (FFD), iced tea, coffee drinks and energy drinks. The mean weekly sweetened beverage load in this population, calculated from the frequency and amount of consumption, was 145 ounces when all categories were considered. When SSB categories were considered independently, sports drinks (45 oz.) had the highest contribution to the mean sweetened beverage load followed by FFD (41 oz.), iced tea (27 oz.), soda (26 oz.) coffee drinks (15 oz.) and energy drinks (2 oz.). Sweetened beverage load was higher in boys (151 oz.) than girls (138 oz.) and was highest in Hispanics (159 oz.) followed by whites (152 oz.), blacks (137 oz.) and others (104 oz.). Behaviors that occurred on a usual basis during SSB consumption included watching TV, eating a family meal, eating salty and fried foods, being on the computer and hanging out with friends. Activities concurrent with sports drink consumption included physical activity behaviors whereas soda and FFD did not. Sports drink and FFD consumption commonly co-occurred with fruit consumption. Multiple SSB categories contribute to the total SSB consumption and the common dietary and activity behaviors are distinct between categories. Several of the concurrent behaviors point to the importance of home beverage availability, and to the influence that parents and peers have on SSB consumption. Identifying and assessing intervention strategies targeted to specific beverage categories could be an important step in behavioral intervention research aimed at reducing added sugar consumption, and ultimately, promote a healthy weight in adolescents. ^

Perlongher y ayahuasca : La antropología como una forma bastarda de realismo

Se propone una reflexión en torno a las diferencias que Perlongher percibe entre la producción de una escritura "en académico" (cuya obligación contractual sustentaba su "frugal manducar") y la expresión poética como forma del éxtasis (como una manera de dar forma a la fuerza de la deriva). Se tomarán como punto de partida los ensayos compilados en Prosa plebeya bajo el rubro "Antropología del éxtasis", escritos entre 1990 y 1991; asimismo, las entrevistas concedidas en la etapa final de su vida (1991-1992), reunidas en Papeles insumisos. Si en la "Introducción" de La prostitución masculina, Perlongher denominó a la antropología como una "ciencia de lo sutil" (contrapuesta a la rigidez técnica de la sociología, la estadística moral), en la última de sus entrevistas, la definió como "una forma bastarda de realismo", claramente relacionada con lo literario. La articulación entre antropología y literatura no es nueva, baste recordar las experiencias de Leiris, Carpentier, Arguedas y, más recientemente, los tanteos de la antropología poética chilena. Relaciono la singularidad perlongheriana a la constitución de territorios afectivos, cartografías de estados de ánimo, a través de una escritura barroca, barrial, barrosa, en la que los avatares de poeta y de profesor cohabitan el mismo escenario

Perlongher y ayahuasca : La antropología como una forma bastarda de realismo

Se propone una reflexión en torno a las diferencias que Perlongher percibe entre la producción de una escritura "en académico" (cuya obligación contractual sustentaba su "frugal manducar") y la expresión poética como forma del éxtasis (como una manera de dar forma a la fuerza de la deriva). Se tomarán como punto de partida los ensayos compilados en Prosa plebeya bajo el rubro "Antropología del éxtasis", escritos entre 1990 y 1991; asimismo, las entrevistas concedidas en la etapa final de su vida (1991-1992), reunidas en Papeles insumisos. Si en la "Introducción" de La prostitución masculina, Perlongher denominó a la antropología como una "ciencia de lo sutil" (contrapuesta a la rigidez técnica de la sociología, la estadística moral), en la última de sus entrevistas, la definió como "una forma bastarda de realismo", claramente relacionada con lo literario. La articulación entre antropología y literatura no es nueva, baste recordar las experiencias de Leiris, Carpentier, Arguedas y, más recientemente, los tanteos de la antropología poética chilena. Relaciono la singularidad perlongheriana a la constitución de territorios afectivos, cartografías de estados de ánimo, a través de una escritura barroca, barrial, barrosa, en la que los avatares de poeta y de profesor cohabitan el mismo escenario

Perlongher y ayahuasca : La antropología como una forma bastarda de realismo

Se propone una reflexión en torno a las diferencias que Perlongher percibe entre la producción de una escritura "en académico" (cuya obligación contractual sustentaba su "frugal manducar") y la expresión poética como forma del éxtasis (como una manera de dar forma a la fuerza de la deriva). Se tomarán como punto de partida los ensayos compilados en Prosa plebeya bajo el rubro "Antropología del éxtasis", escritos entre 1990 y 1991; asimismo, las entrevistas concedidas en la etapa final de su vida (1991-1992), reunidas en Papeles insumisos. Si en la "Introducción" de La prostitución masculina, Perlongher denominó a la antropología como una "ciencia de lo sutil" (contrapuesta a la rigidez técnica de la sociología, la estadística moral), en la última de sus entrevistas, la definió como "una forma bastarda de realismo", claramente relacionada con lo literario. La articulación entre antropología y literatura no es nueva, baste recordar las experiencias de Leiris, Carpentier, Arguedas y, más recientemente, los tanteos de la antropología poética chilena. Relaciono la singularidad perlongheriana a la constitución de territorios afectivos, cartografías de estados de ánimo, a través de una escritura barroca, barrial, barrosa, en la que los avatares de poeta y de profesor cohabitan el mismo escenario

Lealtad de la marca de la bebida "AQUARIUS" = "AQUARIUS" beverage brand loyalty

Desde la disciplina del marketing, esta monografía de investigación pretende estudiar cuál es el nivel de lealtad de la marca de la bebida “Aquarius”. Para ello, se ha aplicado la técnica de la encuesta a una muestra aleatoria de 100 estudiantes de la Facultad de Ciencias de la Comunicación del Campus de Fuenlabrada de la Universidad Rey Juan Carlos de Madrid. Los resultados encontrados en este trabajo indican que “Aquarius” se mantiene todavía relacionada con las bebidas reconstituyentes, con una percepción en torno a la salud, con un alto grado de recomendación, y con un nivel de lealtad relativamente medio. = From the discipline of marketing, this research tries to study which is the level of brand loyalty for the drink “Aquarius”. Thus, it has been applied the survey technique to a random sample of 100 students of the Faculty of Communication Sciences in the Fuenlabrada Campus of the University Rey Juan Carlos (Madrid). The findings of this work indicate that “Aquarius” still stays related to drinks tonics, with a perception around the health, a high degree of recommendation, and with a relatively medium loyalty level.

A Cannabinoid CB1 Receptor Positive Allosteric Modulator Reduces Neuropathic Pain in the Mouse with no Psychoactive Effects

Peer reviewed

Loss of signaling through the G protein, Gz, results in abnormal platelet activation and altered responses to psychoactive drugs

Heterotrimeric G proteins mediate the earliest step in cell responses to external events by linking cell surface receptors to intracellular signaling pathways. Gz is a member of the Gi family of G proteins that is prominently expressed in platelets and brain. Here, we show that deletion of the α subunit of Gz in mice: (i) impairs platelet aggregation by preventing the inhibition of cAMP formation normally seen at physiologic concentrations of epinephrine, and (ii) causes the mice to be more resistant to fatal thromboembolism. Loss of Gzα also results in greatly exaggerated responses to cocaine, reduces the analgesic effects of morphine, and abolishes the effects of widely used antidepressant drugs that act as catecholamine reuptake inhibitors. These changes occur despite the presence of other Giα family members in the same cells and are not accompanied by detectable compensatory changes in the level of expression of other G protein subunits. Therefore, these results provide insights into receptor selectivity among G proteins and a model for understanding platelet function and the effects of psychoactive drugs.

A second-generation vaccine protects against the psychoactive effects of cocaine

The effects of immunization with the second-generation cocaine immunoconjugate GND-keyhole limpet hemocyanin (KLH) or with the anti-cocaine mAb GNC92H2 were assessed in a model of acute cocaine-induced locomotor activity. After i.p. administration of cocaine⋅HCl (15 mg/kg), rats were tested in photocell cages, and stereotypy was rated to determine preimmunization drug response (baseline). Experimental animals were subjected to an immunization protocol with GND-KLH or treated with the mAb GNC92H2. Rats were then challenged with systemic cocaine, and their locomotor responses were again measured. Active immunization with GND-KLH produced a 76% decrease in the ambulatory measure (crossovers) in the experimental group and a 12% increase in the control group compared with baseline values. Also, stereotypic behavior was significantly suppressed in the vaccinated animals. Decreases in both measures were seen in the experimental group on two subsequent challenges. The maximum effect was observed at the time of the second challenge with a dramatic 80% decrease in crossovers. Treatment with GNC92H2 resulted in a 69% decrease in crossovers compared with baseline. This effect persisted across two additional challenges over 11 days with decreases of 46–47%. In contrast, the control group showed increases of up to 28%. Significant differences between groups were observed in the stereotypic measure in all three challenges. The results indicate that these immunopharmacotherapeutic agents have significant cocaine-blockade potential and therefore may offer an effective strategy for the treatment of cocaine abuse.

La vida como proceso de sanación: prácticas chamánicas del alto Amazonas en torno a la ayahuasca en España

Esta investigación describe y analiza prácticas chamánicas del alto Amazonas en torno al uso ceremonial de la ayahuasca en España. Los datos proceden del trabajo de campo multisituado que se desarrolla entre agosto de 2008 y agosto de 2014, principalmente en España, y de manera secundaria en Perú. La descripción de las ceremonias de ayahuasca se centra en la actividad de un conjunto de chamanes, tanto españoles como nativos de las regiones amazónica y andina. Esta actividad se orienta a la sanación de los participantes en las ceremonias, y dicha actividad puede ser analizada como un ejemplo de una racionalidad alternativa a la racionalidad dominante en nuestra sociedad, que se fundamenta en una ontología de tipo dualista. Esta racionalidad alternativa muestra una congruencia entre los medios empleados por los chamanes, y los fines terapéuticos perseguidos. Se define como chamán aquella persona que tiene capacidad para interactuar con el mundo de los ‘espíritus’, y para mediar entre los ‘espíritus’ y un grupo de personas. El término chamán se emplea en este trabajo como una categoría analítica para el estudio transcultural de la actividad de estas personas, y no como una autodenominación de los informantes...