ABSTRACT:A method was developed and validated for the simultaneous determination of dimethyltryptamine and β-carfbolines alkaloids (harmine, harmaline and tetrahydroharmine) in ayahuasca preparations. The alkaloids were extracted by menas of solid phase extraction (C18) and detected by gas Introduction -Ayahuasca is obtained by infusing the pounded stems of Banisteriopsis caapi in combination with the leaves of Psychotria viridis. P. viridis is rich in the psychedelic indole N,N-dimethyltryptamine, whereas B. caapi contains substantial amounts of b-carboline alkaloids, mainly harmine, harmaline and tetrahydroharmine, which are monoamine-oxidase inhibitors. Because of differences in composition in ayahuasca preparations, a method to measure their main active constituents is needed. Objective -To develop a gas chromatographic method for the simultaneous determination of dimethyltryptamine and the main b-carbolines found in ayahuasca preparations. Methodology -The alkaloids were extracted by means of solid phase extraction (C 18 ) and detected by gas chromatography with nitrogen/phosphorous detector. Results -The lower limit of quantification (LLOQ) was 0.02 mg/mL for all analytes. The calibration curves were linear over a concentration range of 0.02-4.0 mg/mL (r 2 > 0.99). The method was also precise (RSD < 10%). Conclusion -A simple gas chromatographic method to determine the main alkaloids found in ayahuasca was developed and validated. The method can be useful to estimate administered doses in animals and humans for further pharmacological and toxicological investigations of ayahuasca.

Ayahuasca is a beverage with psychoactive properties used in religious and ceremonial rituals by some religious groups. The main active components of ayahuasca are dimethyltryptamine and the harmala alkaloids with β-carboline structure acting as monoamine oxidase A inhibitors. This combination produces a pronounced activation of serotonergic pathways and presents potential interaction with other psychotropics. The objective of this study was to investigate the possible interactions between ayahuasca and agents employed in general anesthesia. The pharmacological interactions between ayahuasca and morphine or propofol were evaluated in mice using doses of 12, 120 and 1200 mg/kg (0.1 to 10 times the average dose consumed by humans in religious rituals). Ayahuasca alone showed an antinociceptive effect in the writhing and formalin tests, and intensified the analgesic effect of morphine in the hot plate test. Concerning the pharmacological interactions between ayahuasca and propofol, the results were opposite; ayahuasca intensified the depressant effect of propofol in the rotarod test, but decreased the sleeping time induced by propofol. These set of results showed the occurrence of some interactions between ayahuasca and the drugs morphine and propofol, possibly by both pharmacokinetics and pharmacodynamics mechanisms.

presidente ____________________________ 1 o . examinador ____________________________ 2 o . examinador São Paulo, _________ de _____. V Este trabalho é dedicado à: A meu esposo Humberto pela eterna presença nas horas difíceis e infinita compreensão em todos os momentos A meu pai fonte inesgotável de caráter e inteligência A minha mãe Lina e minhas irmãs Regiane e Mônica pelo amor e compreensão por minhas freqüentes ausências A meu orientador e professor Maurício por compartilhar comigo sua sabedoria e sempre me mostrar o melhor caminho a ser seguido A meus "bebês" que me entendem, me acompanham e são minha inspiração VI Agradecimentos A DEUS que é presença constante em minha vida e me dá forças para não desistir nunca Aos meus mestres, responsáveis por meu aperfeiçoamento e motivação na arte do saber Aos meus pais, que incentivaram e possibilitaram minha vida acadêmica, Ruben, Lina a meus familiares, Regiane, Mônica, Heloísa, Camila, Karen, Claudinei e Luis que de uma forma ou de outra, sempre me mostram que tudo vale a pena A meu professor Mauricio Yonamine, pela persistente presença, ensinamentos, confiança para a realização desse trabalho e também por ser para mim modelo de conduta profissional e pessoal Aos professores da Faculdade de Ciências Farmacêuticas, Dr. Ernani Pinto, Dra. Tânia Marcourakis, Dra. Regina Lúcia pelo apoio e incentivo As professoras Tânia Govato e Alice Chasin por me mostrarem o caminho inicial e ser seguido e sempre acreditaram em mim. Ao Sidnei e Felipe pelo auxílio na síntese dos padrões A Daniela Mendes pela amizade, companheirismo e força A Sandra Donnangelo que não desiste nunca Ao Rodrigo que participou ativamente da formatação desse trabalho VII Aos meus colegas pós-