The anxiolytic-like effect of the alkaloid fraction of the psychedelic plant Mimosa tenuiflora (Willd.) Poir.

The present work investigated the anxiolytic effect of the alkaloid fraction (AF II) from the root bark of Mimosa tenuiflora. Female Swiss mice of 6 weeks old were used in this study. The animals were divided into three groups: 0.9% NaCl (vehicle group, s.c.), AF II (6 mg/kg, s.c.), and diazepam - DZP (5 mg/kg, s.c.). The light/dark box and elevated plus maze tests evaluated the animal anxiety. DMT was identified by the HPLC method. AF II showed significant anxiolytic effects in the two behavioural tests used.

5-MeO-DMT induces sleep-like LFP spectral signatures in the hippocampus and prefrontal cortex of awake rats.

5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) is a potent classical psychedelic known to induce changes in locomotion, behaviour, and sleep in rodents. However, there is limited knowledge regarding its acute neurophysiological effects. Local field potentials (LFPs) are commonly used as a proxy for neural activity, but previous studies investigating psychedelics have been hindered by confounding effects of behavioural changes and anaesthesia, which alter these signals. To address this gap, we investigated acute LFP changes in the hippocampus (HP) and medial prefrontal cortex (mPFC) of freely behaving rats, following 5-MeO-DMT administration. 5-MeO-DMT led to an increase of delta power and a decrease of theta power in the HP LFPs, which could not be accounted for by changes in locomotion. Furthermore, we observed a dose-dependent reduction in slow (20-50 Hz) and mid (50-100 Hz) gamma power, as well as in theta phase modulation, even after controlling for the effects of speed and theta power. State map analysis of the spectral profile of waking behaviour induced by 5-MeO-DMT revealed similarities to electrophysiological states observed during slow-wave sleep (SWS) and rapid-eye-movement (REM) sleep. Our findings suggest that the psychoactive effects of classical psychedelics are associated with the integration of waking behaviours with sleep-like spectral patterns in LFPs.

Iron deficiency in astrocytes alters cellular status and impacts on oligodendrocyte differentiation.

Iron deficiency (ID) has been shown to affect central nervous system (CNS) development and induce hypomyelination. Previous work from our laboratory in a gestational ID model showed that both oligodendrocyte (OLG) and astrocyte (AST) maturation was impaired. To explore the contribution of AST iron to the myelination process, we generated an in vitro ID model by silencing divalent metal transporter 1 (DMT1) in AST (siDMT1 AST) or treating AST with Fe3+ chelator deferoxamine (DFX; DFX AST). siDMT1 AST showed no changes in proliferation but remained immature. Co-cultures of oligodendrocyte precursors cells (OPC) with siDMT1 AST and OPC cultures incubated with siDMT1 AST-conditioned media (ACM) rendered a reduction in OPC maturation. These findings correlated with a decrease in the expression of AST-secreted factors IGF-1, NRG-1, and LIF, known to promote OPC differentiation. siDMT1 AST also displayed increased mitochondrial number and reduced mitochondrial size as compared to control cells. DFX AST also remained immature and DFX AST-conditioned media also hampered OPC maturation in culture, in keeping with a decrease in the expression of AST-secreted growth factors IGF-1, NRG-1, LIF, and CNTF. DFX AST mitochondrial morphology and number showed results similar to those observed in siDMT1 AST. In sum, our results show that ID, induced through two different methods, impacts AST maturation and mitochondrial functioning, which in turn hampers OPC differentiation.

Safety and tolerability of inhaled N,N-Dimethyltryptamine (BMND01 candidate): A phase I clinical trial.

Psychedelics are being increasingly examined for their therapeutic potential in mood disorders. While the acute effects of ayahuasca, psilocybin, and lysergic acid diethylamide (LSD) last over several hours, inhaled N,N-Dimethyltryptamine (DMT) effects last around 10 min, which might provide a cost- and time-effective alternative to the clinical application of oral psychedelics. We aimed at investigating the safety and tolerability of inhaled DMT (BMND01 candidate). We recruited 27 healthy volunteers to receive a first, lower dose and a second, higher dose (5/20 mg, 7.5/30 mg, 10/40 mg, 12.5/50 mg, or 15/60 mg) of inhaled DMT in an open-label, single-ascending, fixed-order, dose-response study design. We investigated subjective experiences (intensity, valence, and phenomenology), physiological effects (blood pressure, heart rate, respiratory rate, blood oxygen saturation, body temperature), biochemical markers (liver, kidney, and metabolic functions), and adverse events during the acute and post-acute effects of DMT. DMT dose-dependently increased intensity, valence and perceptual ratings. There was a mild, transient, and self-limited increase in blood pressure and heart rate. There were no changes in safety blood biomarkers and no serious adverse events. DMT dose-dependently enhanced subjective experiences and positive valence. Inhaled DMT might be an efficient, non-invasive, safe route of administration, which might simplify the clinical use of this substance. This is the first clinical trial to test the effects of inhaled DMT (BMND01 candidate).

Open-label study of consecutive ibogaine and 5-MeO-DMT assisted-therapy for trauma-exposed male Special Operations Forces Veterans: prospective data from a clinical program in Mexico.

Background: Research in psychedelic medicine has focused primarily on civilian populations. Further study is needed to understand whether these treatments are effective for Veteran populations.Objectives: Here, we examine the effectiveness of psychedelic-assisted therapy among trauma-exposed Special Operations Forces Veterans (SOFV) seeking treatment for cognitive and mental health problems in Mexico.Methods: Data were collected from an ibogaine and 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) clinical treatment program for SOFV with a history of trauma exposure. This clinical program collects prospective clinical program evaluation data, such as background characteristics, symptom severity, functioning (e.g., satisfaction with life, posttraumatic stress disorder symptoms, depression symptoms, anxiety symptoms, sleep disturbance, psychological flexibility, disability in functioning, cognitive functioning, neurobehavioral symptoms, anger, suicidal ideation), and substance persisting/enduring effects through online surveys at four timepoints (baseline/pre-treatment, one-, three-, and six-months after treatment).Results: The majority of the sample (n = 86; Mean Age = 42.88, SD = 7.88) were Caucasian (87.2%), non-Hispanic (89.5%), and males (100%). There were significant and large improvements in self-reported PTSD symptoms (p < .001, d = .414), depression (p < .001, d = .275), anxiety (p < .001, d = .276), insomnia severity (p < .001, d = .351), and post-concussive symptoms (p < .001, d = .389) as well as self-reported satisfaction with life (p < .001, d = .371), psychological flexibility (p < .001, d = .313) and cognitive functioning (p < .001, d = .265) from baseline to one-month follow-up.Conclusions: Data suggest combined ibogaine and 5-MeO-DMT assisted therapy has potential to provide rapid and robust changes in mental health functioning with a signal of durable therapeutic effects up to 6-months. Future research in controlled settings is warranted.

A mechanistic insight for the biosynthesis of N,N-dimethyltryptamine: An ONIOM theoretical approach.

Psychoactive natural products are potent serotonergic agonists capable of modulating brain functions such as memory and cognition. These substances have shown therapeutic potential for treating various mental disorders. The fact that N,N-dimethyltryptamine (DMT) is produced endogenously in several plants and animals, including humans, makes it particularly attractive. As an amino acid-derived alkaloid, the DMT biosynthetic pathway is part of the L-tryptophan biochemical cascade and can be divided into the decarboxylation by an aromatic L-amino acid decarboxylase (AADC) for tryptamine formation and the subsequent double-methylation by the indolethylamine-N-methyltransferase (INMT) through the cofactor S-adenosyl-L-methionine (SAM), a methyl donor. Unlike the decarboxylation mechanism of L-tryptophan, the molecular details of the double methylation of tryptamine have not been elucidated. Therefore, we propose an in silico model using molecular dynamics (MD), non-covalent interaction index (NCI) and density functional theory (DFT) calculations with the ONIOM QM:MM B3LYP/6-31+G(d,p):MM/UFF level of theory. Based on the obtained energetic data, the potential energy surface (PES) indicates an SN2 mechanism profile, with the second methylation energy barrier being the rate-limiting step with δG‡=60kJ∙mol-1 larger than the previous methylation, following the NCI analysis showing more repulsive interactions for the second transition state. In addition, the hybridization information of each reaction step provides geometric details about the double-methylation.

Prevalência da esclerose múltipla em pacientes tratados com medicamentos modificadores do curso da doença utilizando dados do Sistema Único de Saúde brasileiro / Prevalence of multiple sclerosis in patients treated with disease-modifying drugs using data from the Brazilian Unified Health System

Objetivo: Compreender o cenário da esclerose múltipla (EM) em relação aos aspectos epidemiológicos, diagnósticos, progressão, tratamento e comorbidades no sistema público brasileiro. Métodos: Trata-se de um estudo retrospectivo e observacional utilizando os sistemas de dados do Departamento de Informática do Sistema Único de Saúde (DataSUS). Os dados utilizados foram obtidos por meio da base de dados do Sistema Ambulatorial/Sistema de Procedimentos de Alta Complexidade (SIA/SUS). Nessa base, o vínculo de registros foi por meio do Cadastro Nacional de Saúde (CNS). Para a quantificação de dados epidemiológicos, foram coletados os dados do Código de Endereçamento Postal (CEP), sexo e data de nascimento, além das bases de geolocalização dos pacientes. Considerou-se como desfecho primário a descrição epidemiológica da população de pacientes em uso ou que usaram MMCD para o tratamento da EM. Como desfecho secundário, consideraram-se as características dos pacientes (gênero, idade, idade ao diagnóstico e comorbidades). Resultados: Foram incluídos na análise 45.011 pacientes. Identificou-se predominância de pacientes do gênero feminino (72,9%) e com idade entre 31 e 60 anos (61,23%) diagnosticados com CID G35 no primeiro registro. A taxa de incidência de pacientes com EM foi maior (2,7 pacientes/100 mil habitantes) na região Sudeste, seguida pela região Sul (2,2 pacientes/100 mil). A prevalência na região Sul teve a maior taxa (18 pacientes/100 mil), seguida pelo Sudeste (16,7 pacientes/100 mil). As betainterferonas e o acetato de glatirâmer foram os medicamentos mais utilizados no primeiro tratamento. O acetato de glatirâmer foi o mais utilizado para o segundo tratamento no período de 2011 até 2017. Em 2018, os MMCD mais utilizados como segundo tratamento foram fingolimode e natalizumabe. A partir de 2019, fingolimode, fumarato de dimetila e natalizumabe foram os medicamentos mais utilizados como segundo tratamento, permanecendo nessa ordem até 2021. Para o terceiro tratamento, o natalizumabe foi o medicamento mais utilizado até 2017. Após esse período, o fingolimode passou a ser mais usado. Desde 2019, com o acesso ao fumarato de dimetila, os medicamentos mais prescritos foram, em ordem decrescente, fingolimode, natalizumabe e fumarato de dimetila. Conclusão: Estabelecer uma análise epidemiológica dos pacientes que usam MMCD para o tratamento da EM no Brasil, além do padrão de tratamento, são dados essenciais para a promoção do tratamento da EM de forma adequada, bem como para a implementação de políticas públicas locais, regionais e nacionais.

Objective: To understand the multiple sclerosis (MS) setting in relation to epidemiological aspects, diagnoses, progression, treatment and comorbidities in the Brazilian public system. Methods: This is a retrospective, observational study using the Unified Health System's Informatics Department [Departamento de Informática do Sistema Único de Saúde] data systems (DataSUS). The data used were obtained by means of the outpatient system/high-complexity procedure system (SIA/SUS) database. In this database, the association of entries was via the National Health Registration [Cadastro Nacional de Saúde] (CNS). For epidemiological data quantification, Zip Code, sex and date of birth information was collected, as well as data from patient geolocation databases. The epidemiological description of the patient population using or having used DMTs for MS treatment was considered as the primary endpoint. Patient characteristics (gender, age, age at diagnosis and comorbidities) were considered as the secondary endpoint. Results: 45,011 patients were included in the analysis. A predominance of female patients (72.9%) aged between 31 and 60 years (61.23%) and diagnosed with ICD code G35 in the first entry was identified. The incidence rate of patients with MS was higher (2.7 patients/100 thousand inhabitants) in the Southeast region, followed by the South region (2.2 patients/100 thousand). Prevalence in the South region had the highest rate (18 patients/100 thousand), followed by the Southeast region (16.7 patients/100 thousand). Interferons beta and glatiramer acetate were the most used drugs in the first treatment. Glatiramer acetate was the most used drug for the second treatment within the period from 2011 to 2017. In 2018, the most common DMTs used as second treatment were fingolimod and natalizumab. From 2019, fingolimod, dimethyl fumarate and natalizumab were the most used drugs as second treatment, remaining in this order until 2021. For the third treatment, natalizumab was the most used drug until 2017. After this period, fingolimod became more widely used. Since 2019, with access to dimethyl fumarate, the most prescribed drugs were, in decreasing order, fingolimod, natalizumab and dimethyl fumarate. Conclusion: Establishing an epidemiological analysis of patients using DMTs for MS treatment in Brazil, in addition to standard of care, results in critical data for adequately promoting MS treatment, as well as for implementing local, regional and national public policies.

Suction Influence on Load-Settlement Curves Predicted by DMT in a Collapsible Sandy Soil.

The plate load test (PLT) is the most reliable in situ testing for studying the load-settlement behaviour of footings on unsaturated collapsible soils. In these soils, the suction profile is not constant along the depth, and the scale effect between the prototype and footing leads to different suction averages and, consequently, different data. One method to eliminate the effect of soil suction on the test data is to fully saturate the soil prior to the test, which is also recommended at the design process for footing on collapsible soils. However, the inundation process on PLTs is expensive and time-consuming, which makes this procedure difficult to incorporate into engineering practice. This study presents a device that can be attached to flat dilatometer (DMT) to allow local inundation of the soil as part of the in situ test campaign and obtain the DMT-constrained modulus (MDMT) for both natural and inundated conditions. The MDMT presented an average reduction of 56% from natural to inundated condition. This parameter can be used in a model to predict load-settlement curves by DMT data considering the suction influence on this behaviour. The curves obtained from the prediction model were compared to curves determined by PLT conducted under the same in situ conditions. Good agreement was found between the curves predicted by DMT and those measured by PLT for both conditions. The proposed procedure, which uses a device attached to the DMT blade, provides an investigation method to obtain the load-settlement curve under different suction conditions, which can help in the selection and performance prediction of shallow foundations, taking into account suction and collapse phenomenon-related problems.

The Association of Classic Serotonergic Psychedelic Use and Intention of Future Use with Nature Relatedness.

This study sought to investigate the effects of different substances on nature relatedness (NR) in the general population. An online cross-sectional survey done in Brazil investigated use of ayahuasca/DMT, psilocybe mushrooms, LSD, MDMA/ecstasy, cocaine, cannabis, and alcohol. NR was assessed using the short-form version of the nature related scale (NR-6). One-way analysis of variance (ANOVA) was used to assess group differences between substance naïve-individuals, past users, and current users of each substance. Regression models were used including all the substances and subsequently, sociodemographic variables. ANOVAs with substances which showed significantly higher NR-6 scores in the regression model were used in order to assess the effect of intention of future use on NR. ANOVAs indicated higher NR in users of classic serotonergic psychedelics (ayahuasca/DMT, psilocybe mushrooms, LSD), cannabis, and MDMA/ecstasy. Regression models showed that current use of ayahuasca/DMT and psilocybe mushrooms, and past use of LSD had a positive association with NR. When sociodemographic variables were added, only ayahuasca/DMT past and current use were positively associated with NR. Intention of future use was only significantly associated with NR in individuals who reported intention of future use of psilocybe mushrooms.

Molecular Pathways of the Therapeutic Effects of Ayahuasca, a Botanical Psychedelic and Potential Rapid-Acting Antidepressant.

Ayahuasca is a psychoactive brew traditionally used in indigenous and religious rituals and ceremonies in South America for its therapeutic, psychedelic, and entheogenic effects. It is usually prepared by lengthy boiling of the leaves of the bush Psychotria viridis and the mashed stalks of the vine Banisteriopsis caapi in water. The former contains the classical psychedelic N,N-dimethyltryptamine (DMT), which is thought to be the main psychoactive alkaloid present in the brew. The latter serves as a source for ß-carbolines, known for their monoamine oxidase-inhibiting (MAOI) properties. Recent preliminary research has provided encouraging results investigating ayahuasca's therapeutic potential, especially regarding its antidepressant effects. On a molecular level, pre-clinical and clinical evidence points to a complex pharmacological profile conveyed by the brew, including modulation of serotoninergic, glutamatergic, dopaminergic, and endocannabinoid systems. Its substances also interact with the vesicular monoamine transporter (VMAT), trace amine-associated receptor 1 (TAAR1), and sigma-1 receptors. Furthermore, ayahuasca's components also seem to modulate levels of inflammatory and neurotrophic factors beneficially. On a biological level, this translates into neuroprotective and neuroplastic effects. Here we review the current knowledge regarding these molecular interactions and how they relate to the possible antidepressant effects ayahuasca seems to produce.

Calcium is a noncompetitive inhibitor of DMT1 on the intestinal iron absorption process: empirical evidence and mathematical modeling analysis.

Iron absorption is a complex and highly controlled process where DMT1 transports nonheme iron through the brush-border membrane of enterocytes to the cytoplasm but does not transport alkaline-earth metals such as calcium. However, it has been proposed that high concentrations of calcium in the diet could reduce iron bioavailability. In this work, we investigate the effect of intracellular and extracellular calcium on iron uptake by Caco-2 cells, as determined by calcein fluorescence quenching. We found that extracellular calcium inhibits iron uptake by Caco-2 cells in a concentration-dependent manner. Chelation of intracellular calcium with BAPTA did not affect iron uptake, which indicates that the inhibitory effect of calcium is not exerted through intracellular calcium signaling. Kinetic studies performed, provided evidence that calcium acts as a reversible noncompetitive inhibitor of the iron transport activity of DMT1. Based on these experimental results, a mathematical model was developed that considers the dynamics of noncompetitive inhibition using a four-state mechanism to describe the inhibitory effect of calcium on the DMT1 iron transport process in intestinal cells. The model accurately predicts the calcein fluorescence quenching dynamics observed experimentally after an iron challenge. Therefore, the proposed model structure is capable of representing the inhibitory effect of extracellular calcium on DMT1-mediated iron entry into the cLIP of Caco-2 cells. Considering the range of calcium concentrations that can inhibit iron uptake, the possible inhibition of dietary calcium on intestinal iron uptake is discussed.

Evidence on the impairing effects of Ayahuasca on fear memory reconsolidation.

RATIONALE: To uncover whether psychedelic drugs attenuate fear memory responses would advance the development of better psychedelic-based treatments for posttraumatic stress disorder (PTSD). Ayahuasca (AYA), a psychedelic brew containing indolamine N, N-dimethyltryptamine (DMT) and ß-carbolines, facilitates fear extinction and improves neural plasticity. Upon retrieval, fear memory undergoes labilization and reconsolidation; however, the effects of AYA on this memory stabilization phase are unknown. OBJECTIVES: We aimed to investigate the effects of AYA treatment on fear memory reconsolidation. METHODS: Fear-conditioned Wistar rats received AYA (60, 120, or 240 mg/kg) or H2O orally via gavage o.g. 20 min before, immediately, or 3 h after a short retrieval session. Analysis of AYA through liquid chromatography-tandem mass spectrometry was used to determine the content of DMT and ß-carbolines in AYA. RESULTS: AYA impaired fear memory reconsolidation when given 20 min before or 3 h after memory retrieval, with the dose of 60 mg/kg being effective at both moments. This dose of AYA was devoid of anxiolytic effect. Importantly, during retrieval, AYA did not change fear expression. The lack of retrieval abolished the reconsolidation impairing effect of AYA. The effects of AYA treatment 20 min before or 3 h after memory retrieval lasted at least 22 days, suggesting no spontaneous recovery of fear memory. Fear memory impairments induced by AYA treatment, at both moments, do not show reinstatement. CONCLUSIONS: Our findings support the view that a low dose of AYA treatment impairs early and late stages of memory reconsolidation instead of facilitating fear extinction.

Ayahuasca for the Treatment of Depression.

Ayahuasca, the vine of the souls in Quechua, is a psychedelic brew with a few formulations that most often include the bark of a liana in the Malpighiaceae family (Banisteriopsis caapi), with leaves from a shrub in the coffee family Rubiaceae (Psychotria viridis). Mixed with water and boiled for hours or days, it produces a brownish-colored liquid with a strong and characteristic taste. Ayahuasca contains the psychedelic tryptamine N,N-Dimethyltryptamine (DMT), and Monoamine Oxidase Inhibitors (MAOi), and in the past few years, it has been tested. In recent years its antidepressant properties have been put to the test. Evidence from open and randomized placebo-controlled clinical trials has shown encouraging results, indicating significant and rapid antidepressant effects, starting as early as 1 day after the ayahuasca intervention. In addition, we have explored the nature of these effects using multivariate measures. In this article, we will review the history, pharmacology, clinical trials, and clinical and behavioral markers associated with the antidepressant effects of ayahuasca.

Baseline Power of Theta Oscillations Predicts Mystical-Type Experiences Induced by DMT in a Natural Setting.

N,N-Dimethyltryptamine (DMT) is a classic psychedelic capable of inducing short-lasting but profound changes in consciousness. As with other psychedelics, the experience induced by DMT strongly depends upon contextual factors, yet the neurobiological determinants of this variability remain unknown. The present study investigated changes in neural oscillations elicited by inhaled DMT, and whether baseline electroencephalography (EEG) recordings could predict the subjective effects reported by the participants. Healthy volunteers (N = 35) were measured with EEG before and during the acute effects of DMT consumed in a natural setting. Source-localized neural oscillations were correlated with the results of multiple questionnaires employed to assess the subjective effects of the drug. DMT resulted in a marked reduction of alpha and beta oscillations, and increased posterior spectral power in the delta, theta and gamma bands. The power of fronto-temporal theta oscillations was inversely correlated with scales indexing feelings of unity and transcendence, which are an integral part of the phenomenology of mystical-type experiences. The robustness of these results was supported using a machine learning model for regression trained and tested following a cross-validation procedure. These results are consistent with the observation that the state of mind prior to consuming a psychedelic drug influences the ensuing subjective experience of the user. They also suggest that baseline EEG screenings before administration of a serotonergic psychedelic could be useful to estimate the likelihood of inducing mystical-type experiences, previously linked to sustained positive effects in well-being and improved outcome of therapeutic interventions.

Psychosis induced by abuse of ayahuasca: a case report / Psicosis inducida por el abuso de la ayahuasca: un caso clínico

A B S T R A C T Ayahuasca is a psychotropic infusion prepared by boiling the bark of Amazonian plants and has many psychopharmacological effects not fully understood. Some of those effects are used as treatment for different diseases. However, the side effects of ayahuasca, including ayahuasca-induced psychosis, are an important issue. Here we report the case of a patient who had a psychotic episode after taking ayahuasca and who was successfully treated with antipsychotic medication. Given the current spread of ayahuasca consumption in developed societies, the present case highlights the need for better understanding and regulation of the social-legal condition of ayahuasca and the need for further research. Additionally, psycho-education seems advisable in order to create awareness of the potential risks of the use of ayahuasca.

RESUMEN La ayahuasca es una bebida psicotrópica preparada a través de la cocción de plantas de la cuenca amazónica que tiene muchos efectos psicofarmacológicos no del todo estudiados. Algunos de esos efectos son usados como tratamiento de diversas patologías. Sin embargo, existen efectos secundarios de la ayahuasca que deben ser tenidos en cuenta, entre ellos psicosis inducida por ayahuasca. Reportamos un caso de un paciente que, tras autoadministración de ayahuasca, presentó un episodio psicótico y que fue satisfactoriamente tratado con antipsicóticos. Dada el uso cada vez más frecuente de ayahuasca en las sociedades desarrolladas, el caso actual resalta las necesidades de entender, regular e investigar el uso de la ayahuasca. Además, crear conciencia de los potenciales riesgos del uso de ayahuasca a través de la psicoeducación debería ser implementado.

Fast Hollow Fiber Liquid-Phase Microextraction as a Greener Alternative for the Determination of N,N-Dimethyltryptamine and Harmala Alkaloids in Human Urine.

Ayahuasca tea is an entheogen hallucinogenic beverage used for shamanic and spiritual purposes, prepared by the decoction of different Amazonian plants containing N,N-dimethyltryptamine (DMT) and harmala alkaloids. Since the therapeutic potential of this tea has been broadly studied in recent years, mainly for the treatment of psychiatric disorders, the determination of the ayahuasca tea components in human and animal matrices is of utmost importance. In order to avoid the use of large amounts of toxic solvents, typically employed in traditional sample preparation methods, hollow fiber liquid-phase microextraction (HF-LPME) presents a greener and time-saving alternative. The present study aims to fully develop and apply an HF-LPME method for the determination of DMT, harmine (HRM), harmaline (HRL), and tetrahydroharmine (THH) in human urine samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fractional factorial and Box-Behnken designs were used to identify and optimize significant method variables. Once optimized, validation has shown a limit of detection (LoD) of 1.0 ng/ml for DMT and 2.0 ng/ml for the harmala alkaloid. The limit of quantification (LoQ) was of 5.0 ng/ml for all analytes. The method has shown to be linear over a concentration range of 5-200 ng/ml (r 2 ≥ 0.99). Intra/inter-day precision and accuracy met the acceptance criteria at the three quality control (QC) levels studied (15.0, 90.0, and 170.0 ng/ml, n = 6, each). Matrix effect evaluation showed predominant ion enhancement and recovery values were above 80%. Dilution factors of 10- and 20-fold have shown acceptable values of accuracy. Selectivity studies showed no interferences. Analysis of eight authentic samples collected from four subjects proved method feasibility. A simple, time-saving and green alternative for the analysis of DMT and harmala alkaloids in human urine samples was developed, optimized using design of experiments, fully validated and applied to authentic samples.

Biodiversity of ß-Carboline Profile of Banisteriopsis caapi and Ayahuasca, a Plant and a Brew with Neuropharmacological Potential.

Ayahuasca is a psychoactive infusion with a large pharmacological application normally prepared with Banisteriopsis caapi, which contains the monoamine oxidase inhibitors ß-carbolines, and Psichotria virids, which contains the serotonin receptor agonist N,N dimethyltryptamine (DMT). The objectives of this study were to investigate the chemical profile of B. caapi and of ayahuasca collected in various Brazilian regions. In total, 176 plant lianas, of which 159 B. caapi and 33 ayahuasca samples were analyzed. Dried liana samples were powdered, extracted with methanol, diluted, and analyzed by LC-MS/MS. Ayahuasca samples were diluted and analyzed. Mean concentrations in B. caapi were 4.79 mg/g harmine, 0.451 mg/g harmaline, and 2.18 mg/g tetrahydroharmine (THH), with a high variability among the samples (RSD from 78.9 to 170%). Native B. caapi samples showed significantly higher harmine concentrations than cultivated ones, and samples from the Federal District/Goiás had higher THH content than those collected in the State of Acre. The other Malpighiaceae samples did not contain ß-carbolines, except for one D. pubipetala sample. Concentrations in ayahuasca samples ranged from 0.109 to 7.11 mg/mL harmine, 0.012 to 0.945 mg/mL harmaline, 0.09 to 3.05 mg/mL THH, and 0.10 to 3.12 mg/mL DMT. The analysis of paired ayahuasca/B. caapi confirmed that harmine is reduced to harmaline and to THH during the brew preparation. This is the largest study conducted with Malpighiaceae samples and showed a large variability in the main ß-carbolines present in B. caapi. This biodiversity is a challenge for standardization of the material used in ethnopharmacological studies of B. caapi and ayahuasca.

A rapid analytical strategy for the determination of ayahuasca alkaloids in non-ritualistic approaches by UHPLC-MS/MS.

INTRODUCTION: Ayahuasca is a beverage composed by a mixture of herbs which contain the compound N,N-dimethyltriptamine (DMT) and the ß-carbolines. Although its use is legalized in Brazil only for religious and spiritual ceremonies, there is a growing black market specialized in the distribution of these compounds in form of herbal material through internet and mail. The purpose of this work was the development of an ultra-high-performance liquid chromatography-tandem mass spectrometry method for the determination of ayahuasca alkaloids and its application in seized ayahuasca products. METHODS: An aliquot of seized products was weighted and diluted with methanol. An aliquot of this solution was added with internal standard (DMT-d6), followed by injection in the analytical system. RESULTS: The limit of quantitation was 10ng/mL for DMT and 25ng/mL for harmine, harmaline and tetrahydroharmine. The concentration ranges used were 10-100ng/mL for DMT, harmine and harmaline and all analytes presented a coefficient of determination (r2)≥0,99. Analysis of four seized samples presented concentrations of DMT ranging between 31.5 and 46.5mg/g. Presence of ß-carbolines was not detected in the products. The variability of DMT concentrations can be correlated with the potential intoxications described in the literature. CONCLUSION: This work successfully established a determination method for ayahuasca alkaloids in herbal material. In addition, the workflow proved to be simple, rapid and useful to estimate the concentration of psychoactive compounds in seized materials, leading to further investigation of ayahuasca ritualistic or recreational exposure.

Stability Evaluation of DMT and Harmala Alkaloids in Ayahuasca Tea Samples.

Ayahuasca tea is a hallucinogenic beverage used for religious purposes in Brazil and many other countries that has therapeutic potential in the treatment of some mental health disorders. In the context of psychedelic research, quantification of the tea's main alkaloids prior to its administration in animal or human studies is essential. For this reason, this study aims to provide information regarding the stability of the main ayahuasca alkaloids (dimethyltryptamine, DMT; harmine, HRM; tetrahydroharmine, THH; harmaline, HRL) in three different conditions: (1) A year stored in a refrigerator either in plastic or glass containers, (2) seven days at 37 °C to reproduce usual mail transportation, and (3) after three freeze-thaw cycles. Samples were quantified after a dilute-and-shoot procedure using liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS). There was no significant degradation of DMT concentration over time in all tested conditions. Harmala alkaloids (THH, HRL, and HRM) showed important variations after long-term and high-temperature storages. Although DMT has proven to be stable in all studied conditions, the harmala alkaloids revealed intense degradation and even concentration increment. This may be caused by degradation, alkaloid inter-conversion, and leaching from tea precipitate material. Therefore, ayahuasca quantification before administration in controlled sets is mandatory.

Investigation of Ayahuasca ß-Carboline Alkaloids and Tryptamine in Sweat Samples from Religious Community Participants by GC-MS.

Ayahuasca, a hallucinogenic beverage used in religious rituals in South America, has become a global phenomenon. Its main active components are the ß-carbolines alkaloids, harmine (HRM) and harmaline (HRL), as well as the potent hallucinogen N,N-dimethyltryptamine (DMT). Despite its rising consumption, information regarding possible clinical applications and toxicological effects of ayahuasca is still limited. This study presents the first investigation of the use of sweat for the determination of DMT, HRM and HRL in ayahuasca users during a religious ritual. Sweat is an alternative matrix with advantages over many conventional biological samples, mainly because the collection procedure is non-invasive, easy and simple and samples can be collected without disturbing the religious ritual. In the study, solid-phase extraction was performed under basic conditions. Linearity was observed ranging from 20 to 1500 ng/patch with coefficients of determination (R2) higher than 0.99 for all analytes. The results indicated high selectivity for all investigated analytes, with extraction efficiency exceeding 70%, accuracy ranging from 87.5 to 102.4%, intra-assay precision of 1.85-9.44% and inter-assay precision between 3.34 and 9.85%. The limits of detection were 15 ng/patch for HRM and HRL and 10 ng/patch for DMT. The sweat proved to be a viable option to monitor ayahuasca use.