3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") 3' is a 'psychedelic amphetamine' that has gained popularity over the past 20 years because of its ability to produce strong feelings of comfort, empathy, and connection to others. MDMA analysis in blood and urine samples have been consistently used for clinical pharmacology studies and forensic science cases. However, new developments in clinical toxicology require new analytical approaches and the use of alternative biological matrices for establishing whether individuals have consumed the drug, when and/or if they have been acting under the effect of the drug.It is postulated that MDMA physic-chemical properties: (i) pKa of around 9.9 corresponding to a weak base that facilitates the transfer of MDMA from plasma (pH=7.4) to fluids/matrices with a favourable pH gradient, (ii) high liposolubility with volumes of distribution between 6 and 7 liters per kilogram, (iii) low protein binding, favour its distribution to biological matrices in humans. Several non-conventional biological matrices such as hair, sweat and saliva, because of drug accumulation due to its physico-chemical properties, might be of use for the detection of past and recent exposure to MDMA.Three different studies were set-up. The study 1 investigating the pharmacokinetics of MDMA in saliva after a single oral dose administration of 100 mg to eight healthy volunteers, the second investigating the pharmacokinetics of MDMA in sweat after a single dose administration of 100 mg to eight healthy volunteers, and finally a study on segmental analysis of MDMA in hair of thirteen drug consumers with different patterns of consumption..The first study evidenced that MDMA is excreted in saliva, after a single 100 mg dose administration, with concentrations (range 1728.9-6510 µg/ml at 1.5 h after drug intake) one order of magnitude higher than those observed in plasma (range 134.9-223 µg/ml at 1.5 h after drug intake) and following a time course kinetics which parallels that of plasma and that of subjective effects and psychomotor performance. On-site testing by Drugwipe device proved suitable to detect individuals under the influence of drug effects in the first 6 hours after drug intake by non-invasive and rapid collection of salivary specimens.The second study showed that MDMA appears in sweat and can be quantified already in the first few hours after a single dose administration, when subjective effects are apparent (concentration range 3.2-1326 ng/pacth). This result makes:the sweat patch technology useful for monitoring MDMA accumulation in sweat at least during the 24 hours after a single administration, On-site sweat testing by drugwipe device suitable to detect individuals under the influence of drug effects by non-invasive and rapid collection of minute amounts of sweat.MDMA appears in hair from consumers (concentration range 1.2-12.6 ng/mg hair) and can be detected in hair segments corresponding to the last one, six and twelve month of repeated drug use. For this reason:Hair analysis of MDMA can be used to evaluate exposure or abstinence to the drug in the last months, hair concentration of MDMA in different hair segments can predict levels of drug use(r2=0.92) and can be eventually associated to chronic psychophysical effects induced by the repeated drug use.The measurement of MDMA in saliva is a valuable alternative to determination of plasma drug concentrations both in clinical and toxicological studies A common characteristic of the three different matrices is that the parent drug MDMA was always the principal, most abundant analyte detected, whose concentration could be associated with drug-induced effects and drug history. As already assessed, drug analysis in hair extends the information of drug consumption to a wider time-window than that of other non-invasive biological matrices, such as saliva and sweat. These latter two matrices can account for acute pharmacological effects induced by the drug, while results from hair testing can be used to assess repeated exposure to drug and eventual association with long term drug induced effects, such as neurotoxicity and psychological performance in the specific case of MDMA.

O consumo abusivo de substâncias psicoativas é um problema de saúde pública, sendo as análises toxicológicas uma importante ferramenta para seu controle. Atualmente, o cabelo está entre as mais utilizadas matrizes biológicas para análise de substâncias psicoativas, devido principalmente ao seu amplo período de deteção. As análises toxicológicas de drogas de abuso podem ser conduzidas através de técnicas de triagem seguidas por técnicas confirmatórias para os resultados positivos. No presente trabalho, foi avaliada a adaptação do método de triagem para substâncias psicoativas em cabelo a partir do kit para imunoensaio em sangue DOA I WB P com a tecnologia Biochip - Randox Laboratórios®. As análises foram realizadas para as classes das anfetaminas, benzodiazepínicos, barbitúricos, cocaína, opiáceos e canabinoides. Os ensaios imunológicos seguiram a partir da análise de amostras reais, adicionadas e artificiais sendo todas previamente confirmadas por técnica de cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). Pelos resultados obtidos pôde-se observar que o kit não apresenta capacidade de analisar a classe de canabinoides. Já para os opiáceos, barbitúricos e anfetamina a aplicação do imunoensaio se mostrou promissora, entretanto a falta de amostras reais positivas impossibilitou a validação do método. Para cocaína, possíveis fontes de contaminação podem ter alterado os resultados impossibilitando a sua avaliação. Benzodiazepínicos, MDMA e metanfetamina apresentaram efeito matriz significativo, não havendo diferenciação entre amostras positivas e negativas. Desta forma, a validação e aplicação do método de imunoensaio para amostras de cabelo não foi possível devido ao elevado efeito de matriz apresentado pelo kit e a falta de amostras reais positivas impossibilitando a validação de parâmetros como sensibilidade, especificidade e exatidão. / The abuse of psychoactive substances is a public health problem and toxicological analysis is an important tool for its control. Currently, hair is among the most widely used biological matrices psychoactive substance analysis, mainly for its detection window. The toxicological analysis of drugs of abuse may be conduted by screening techniques followed by confirmation techniques for positive results. In the present work, an adaptation of a screening method for psychoactive substances in hair was evaluated from the DOA I WB P blood immunoassay kit with a Biochip - Randox Laboratories® technology. The analysis were performed for the drug classes of amphetamines, benzodiazepines, barbiturates, cocaine, opiates and cannabinoids. The immunological assays followed the analysis of real, spiked and artificial samples that have been previously confirmed by gas chromatography coupled to mass spectrometry (GC-MS) technique. From the results obtained it was observed that the kit does not sufficient capacity for analysis of cannabinoids. As for opiates, barbiturates and amphetamines, the immunoassay application proved to be promising. However, a lack of real samples made it impossible to validate the method. For cocaine, finding sources of contamination may have altered the results, making it impossible to evaluate. Benzodiazepines, MDMA and methamphetamine showed significant matrix effect, with no difference between positive and negative samples. Thus, a validation and application of the immunoassay method for hair samples was not possible due to the high matrix effect for the kit and a lack of actual samples making it extremely difficult to validate parameters such as sensitivity, specificity and accuracy.

O consumo abusivo de substâncias psicoativas é um problema de saúde pública, sendo as análises toxicológicas uma importante ferramenta para seu controle. Atualmente, o cabelo está entre as mais utilizadas matrizes biológicas para análise de substâncias psicoativas, devido principalmente ao seu amplo período de deteção. As análises toxicológicas de drogas de abuso podem ser conduzidas através de técnicas de triagem seguidas por técnicas confirmatórias para os resultados positivos. No presente trabalho, foi avaliada a adaptação do método de triagem para substâncias psicoativas em cabelo a partir do kit para imunoensaio em sangue DOA I WB P com a tecnologia Biochip - Randox Laboratórios®. As análises foram realizadas para as classes das anfetaminas, benzodiazepínicos, barbitúricos, cocaína, opiáceos e canabinoides. Os ensaios imunológicos seguiram a partir da análise de amostras reais, adicionadas e artificiais sendo todas previamente confirmadas por técnica de cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). Pelos resultados obtidos pôde-se observar que o kit não apresenta capacidade de analisar a classe de canabinoides. Já para os opiáceos, barbitúricos e anfetamina a aplicação do imunoensaio se mostrou promissora, entretanto a falta de amostras reais positivas impossibilitou a validação do método. Para cocaína, possíveis fontes de contaminação podem ter alterado os resultados impossibilitando a sua avaliação. Benzodiazepínicos, MDMA e metanfetamina apresentaram efeito matriz significativo, não havendo diferenciação entre amostras positivas e negativas. Desta forma, a validação e aplicação do método de imunoensaio para amostras de cabelo não foi possível devido ao elevado efeito de matriz apresentado pelo kit e a falta de amostras reais positivas impossibilitando a validação de parâmetros como sensibilidade, especificidade e exatidão. / The abuse of psychoactive substances is a public health problem and toxicological analysis is an important tool for its control. Currently, hair is among the most widely used biological matrices psychoactive substance analysis, mainly for its detection window. The toxicological analysis of drugs of abuse may be conduted by screening techniques followed by confirmation techniques for positive results. In the present work, an adaptation of a screening method for psychoactive substances in hair was evaluated from the DOA I WB P blood immunoassay kit with a Biochip - Randox Laboratories® technology. The analysis were performed for the drug classes of amphetamines, benzodiazepines, barbiturates, cocaine, opiates and cannabinoids. The immunological assays followed the analysis of real, spiked and artificial samples that have been previously confirmed by gas chromatography coupled to mass spectrometry (GC-MS) technique. From the results obtained it was observed that the kit does not sufficient capacity for analysis of cannabinoids. As for opiates, barbiturates and amphetamines, the immunoassay application proved to be promising. However, a lack of real samples made it impossible to validate the method. For cocaine, finding sources of contamination may have altered the results, making it impossible to evaluate. Benzodiazepines, MDMA and methamphetamine showed significant matrix effect, with no difference between positive and negative samples. Thus, a validation and application of the immunoassay method for hair samples was not possible due to the high matrix effect for the kit and a lack of actual samples making it extremely difficult to validate parameters such as sensitivity, specificity and accuracy.