Harmala Alkaloids Identify Ayahausca Intoxication in a Urine Drug Screen.

BACKGROUND: Ayahausca is an ethnobotanical drink of South America and the compound dimethyltryptamine (DMT) is primarily responsible for the hallucinogenic effects. DMT has a short half-life and its detection in urinary drug screens is challenging. We investigate a simple alternate approach to detect ayahuasca consumption by relying on other constituents of the drink, the ß-carboline harmala alkaloids. METHODS: Three commercially sourced harmala alkaloids were characterized and added to a non-targeted high-resolution mass spectrometry urine drug screening method. All analyses were performed on a Waters Xevo G2-XS LC-QTof, in positive electrospray ionization mode. The mass detector was operated in MSE mode and data processed with UNIFI™ software. A urine specimen from a patient suspected to have consumed ayahuasca was analyzed by a non-targeted drug screen. RESULTS: The harmala alkaloids: harmine, harmaline and tetrohydroharmaline (THH) were characterized and their detection data added to the toxicology screening library. Harmaline and THH were detected in the patient's urine specimen. CONCLUSION: The inclusion of the harmala alkaloids into the drug screen method library may enable the detection of ayahuasca use in patients that undergo non-targeted drug screen.

Comparing the Detection of Endogenous Psychedelics in Individuals With and Without Alleged Mediumistic Experiences.

CONTEXT: Mediumship is the alleged ability to communicate with deceased personalities. Previous studies have suggested that the endogenous psychotomimetic molecules bufotenine (BT) and dimethyltryptamine (DMT) may play a role in the pathogenesis of psychotic disorders. Distortion of perceptions observed during spiritual experiences could supposedly relate to these substances. OBJECTIVE: To compare the presence of BT and DMT in human urine samples between individuals with and without mediumistic experiences. METHODS: All participants (5 from medium's group - MG and 5 from non-medium's group - CG) undertook a single night continuous 6-h urine pool collection (6:00-11:59 PM). Mediums collected urine samples in nights when they reported having experienced mediumistic communication. A sensitive high-performance liquid chromatography-mass spectrometry (HPLC-MS) assay was used. Questionnaires were used to detect common mental disorders symptoms, and to screen and quantify anomalous experiences. RESULTS: DMT was not detected in any urine specimen tested. The presence of BT detection in urine samples was greater in CG (2/5) than in MG (1/5), with no significant differences (p > 0.99). MG reported more anomalous experiences than CG (6.6±0.8 vs. 2.2±1.5, p = 0.03), but there was no difference concerning their mental health. CONCLUSION: There were no differences between individuals with and without alleged mediumistic experiences concerning endogenous psychedelics. Both BT and DMT are highly sensitive to metabolism by monoamine oxidase and to N-oxidation, and do not survive in the periphery for long. Alternative strategies should be considered to further investigate the putative role of the endogenous psychedelics pathway for the spiritual experiences.

Metabolism and urinary disposition of N,N-dimethyltryptamine after oral and smoked administration: a comparative study.

N,N-dimethyltryptamine (DMT) is a widely distributed plant alkaloid that displays partial agonist activity at the 5-HT2A receptor and induces intense psychedelic effects in humans when administered parenterally. However, self-administration studies have reported a total lack of activity following oral intake. This is thought to be due to extensive degradation by monoamine oxidase (MAO). Despite increased use of DMT and DMT-containing preparations, such as the plant tea ayahuasca, the biotransformation of DMT in humans when administered alone is relatively unknown. Here we used high performance liquid chromatography (HPLC)/electrospray ionization (ESI)/selected reaction monitoring (SRM)/tandem mass spectrometry (MS/MS) to characterize the metabolism and disposition of oral and smoked DMT. Twenty-four-hour urine samples were obtained from 6 DMT users before and after intake of 25 mg DMT doses on two separate sessions. In one session, DMT was taken orally and in another it was smoked. After oral ingestion, no psychotropic effects were experienced and no DMT was recovered in urine. MAO-dependent indole-3-acetic acid (IAA) represented 97% of the recovered compounds, whereas DMT-N-oxide (DMT-NO) accounted for only 3%. When the smoked route was used, the drug was fully psychoactive, unmetabolized DMT and DMT-NO rose to 10% and 28%, respectively, and IAA levels dropped to 63%. An inverse correlation was found between the IAA/DMT-NO ratio and subjective effects scores. These findings show that in the smoked route a shift from the highly efficient MAO-dependent to the less efficient CYP-dependent metabolism takes place. This shift leads to psychoactivity and is analogous to that observed in ayahuasca preparations combining DMT with MAO inhibitors.

Metabolism and disposition of N,N-dimethyltryptamine and harmala alkaloids after oral administration of ayahuasca.

Ayahuasca is an Amazonian psychotropic plant tea obtained from Banisteriopsis caapi, which contains ß-carboline alkaloids, chiefly harmine, harmaline and tetrahydroharmine. The tea usually incorporates the leaves of Psychotria viridis or Diplopterys cabrerana, which are rich in N,N-dimethyltryptamine (DMT), a psychedelic 5-HT(2A/1A/2C) agonist. The ß-carbolines reversibly inhibit monoamine-oxidase (MAO), effectively preventing oxidative deamination of the orally labile DMT and allowing its absorption and access to the central nervous system. Despite increased use of the tea worldwide, the metabolism and excretion of DMT and the ß-carbolines has not been studied systematically in humans following ingestion of ayahuasca. In the present work, we used an analytical method involving high performance liquid chromatography (HPLC)/electrospray ionization (ESI)/selected reaction monitoring (SRM)/tandem mass spectrometry(MS/MS) to characterize the metabolism and disposition of ayahuasca alkaloids in humans. Twenty-four-hour urine samples were obtained from 10 healthy male volunteers following administration of an oral dose of encapsulated freeze-dried ayahuasca (1.0 mg DMT/kg body weight). Results showed that less than 1% of the administered DMT dose was excreted unchanged. Around 50% was recovered as indole-3-acetic acid but also as DMT-N-oxide (10%) and other MAO-independent compounds. Recovery of DMT plus metabolites reached 68%. Harmol, harmalol, and tetrahydroharmol conjugates were abundant in urine. However, recoveries of each harmala alkaloid plus its O-demethylated metabolite varied greatly between 9 and 65%. The present results show the existence in humans of alternative metabolic routes for DMT other than biotransformation by MAO. Also that O-demethylation plus conjugation is an important but probably not the only metabolic route for the harmala alkaloids in humans.

A critical review of reports of endogenous psychedelic N, N-dimethyltryptamines in humans: 1955-2010.

Three indole alkaloids that possess differing degrees of psychotropic/psychedelic activity have been reported as endogenous substances in humans; N,N-dimethyltryptamine (DMT), 5-hydroxy-DMT (bufotenine, HDMT), and 5-methoxy-DMT (MDMT). We have undertaken a critical review of 69 published studies reporting the detection or detection and quantitation of these compounds in human body fluids. In reviewing this literature, we address the methods applied and the criteria used in the determination of the presence of DMT, MDMT, and HDMT. The review provides a historical perspective of the research conducted from 1955 to 2010, summarizing the findings for the individual compounds in blood, urine, and/or cerebrospinal fluid. A critique of the data is offered that addresses the strengths and weaknesses of the methods and approaches to date. The review also discusses the shortcomings of the existing data in light of more recent findings and how these may be overcome. Suggestions for the future directions of endogenous psychedelics research are offered.

In vivo long-term kinetics of radiolabeled n,n-dimethyltryptamine and tryptamine.

UNLABELLED: N,N-dimethyltryptamine (DMT), a strong psychodysleptic drug, has been found in higher plants, shamanic hallucinogenic beverages, and the urine of schizophrenic patients. The aim of this work was to gain better knowledge on the relationship between this drug and hallucinogenic processes by studying DMT behavior in comparison with tryptamine. METHODS: (131)I-labeled DMT and tryptamine were injected into rabbits. γ-Camera and biodistribution studies were performed. Brain uptake, plasma clearance, and renal excretion were assessed for each indolealkylamine. RESULTS: DMT and tryptamine showed different behavior when brain uptake, residence time, and excretion were compared. Labeled DMT entered the brain 10 s after injection, crossed the blood-brain barrier, and bound to receptors; then it was partially renally excreted. It was detected in urine within 24 h after injection and remained in the brain, even after urine excretion ceased; up to 0.1% of the injected dose was detected at 7 d after injection in the olfactory bulb. In contrast, tryptamine was rapidly taken up in the brain and fully excreted 10 min after injection. CONCLUSION: To our knowledge, this is the first demonstration that exogenous DMT remains in the brain for at least 7 d after injection. Although labeled DMT and tryptamine behave as agonists for at least 5-hydroxytryptamine 2A receptor, 5-hydroxytryptamine 2C receptor, trace amine-associated receptor, and σ-1 putative receptor targets, binding to the latter can explain the different behavior of labeled DMT and tryptamine in the brain. The persistence in the brain can be further explained on the basis that DMT and other N,N-dialkyltryptamines are transporter substrates for both the plasma membrane serotonin transporter and the vesicle monoamine transporter 2. Furthermore, storage in vesicles prevents DMT degradation by monoamine oxidase. At high concentrations, DMT is taken up by the serotonin transporter and further stored in vesicles by the vesicle monoamine transporter 2, to be released under appropriate stimuli. Moreover, the (131)I-labeling proved to be a useful tool to perform long-term in vivo studies.

Methodology for and the determination of the major constituents and metabolites of the Amazonian botanical medicine ayahuasca in human urine.

Ayahuasca, also known as caapi or yage among various South American groups, holds a highly esteemed and millennia-old position in these cultures' medical and religious pharmacopeia. There is now an increasing interest in the potential for modern medical applications of ayahuasca, as well as concerns regarding its increasing potential for abuse. Toxicological and clinical research to address these issues will require information regarding its metabolism and clearance. Thus, a rapid, sensitive and specific method for characterization and quantitation of the major constituents and of the metabolites of ayahuasca in urine is needed. The present research provides a protocol for conducting such analyses. The characteristics of the method, conducted by sample dilution and using HPLC-electrospray ionization (ESI)-selected reaction monitoring (SRM)-tandem mass spectrometry, are presented. The application of the analytical protocol to urine samples collected from three individuals that were administered ayahuasca has also been demonstrated. The data show that the major metabolite of the hallucinogenic component of ayahuasca, N,N-dimethyltryptamine (DMT), is the corresponding N-oxide, the first time this metabolite has been described in in vivo studies in humans. Further, very little DMT was detected in urine, despite the inhibition of monoamine oxidase afforded by the presence of the harmala alkaloids in ayahuasca. The major harmala alkaloid excreted was tetrahydroharmine. Other excretion products and metabolites were also identified and quantified. The method described would be suitable for use in further toxicological and clinical research on ayahuasca.

Urinary excretion of dietary contaminants in horses.

REASONS FOR PERFORMING STUDY: Presence of drugs is completely prohibited in post racing urine samples by most of racing and competition authorities, even if environmental contamination might occur. OBJECTIVES: To assess the daily dose of several contaminants absorbed through the diet that would result in detectable concentrations in urine. METHODS: Caffeine, theobromine, theophylline, atropine, scopolamine, bufotenine, DMT or morphine were administered orally to 6 horses, in different dosages, for 3 days before their urine was sampled for regular anti-doping tests. RESULTS: Theobromine, theophylline, bufotenine and morphine daily intake >10 mg, 2 mg, 10 mg and 200 microg, respectively, by a performance horse, were found to result in detectable urinary concentrations. At the 2 tested doses, atropine (5 and 15 mg) and dimethyltryptamine (3 and 10 mg) were not detected in urine. For caffeine and scopolamine, even the lowest dosage tested (5 mg/horse/day and 2 mg/horse/day respectively) induced detectable concentrations of the molecule in urine. CONCLUSIONS: Horses fed dietary contaminants, even at level much below the effective dosage, may be positive to antidoping urine analysis. Further research is needed to gain more confident results on a daily safe intake for caffeine and scopolamine. POTENTIAL RELEVANCE: Selection of feed materials appears to be of great importance to prevent non voluntary positive result to anti-doping tests.

Potentially hallucinogenic 5-hydroxytryptamine receptor ligands bufotenine and dimethyltryptamine in blood and tissues.

Bufotenine and N,N-dimethyltryptamine (DMT) are hallucinogenic dimethylated indolethylamines (DMIAs) formed from serotonin and tryptamine by the enzyme indolethylamine N-methyltransferase (INMT) ubiquitously present in non-neural tissues. In mammals, endogenous bufotenine and DMT have been identified only in human urine. The DMIAs bind effectively to 5HT receptors and their administration causes a variety of autonomic effects, which may reflect their actual physiological function. Endogenous levels of bufotenine and DMT in blood and a number of animal and human tissues were determined using highly sensitive and specific quantitative mass spectrometric techniques. A new finding was the detection of large amounts of bufotenine in stools, which may be an indication of its role in intestinal function. It is suggested that fecal and urinary bufotenine originate from epithelial cells of the intestine and the kidney, respectively, although the possibility of their synthesis by intestinal bacteria cannot be excluded. Only small amounts of the DMIAs were found in somatic or neural tissues and none in blood. This can be explained by rapid catabolism of the DMIAs by mitochondrial monoamino-oxidase or by the fact that the dimethylated products of serotonin and tryptamine are not formed in significant amounts in most mammalian tissues despite the widespread presence of INMT in tissues.

Determination of potentially hallucinogenic N-dimethylated indoleamines in human urine by HPLC/ESI-MS-MS.

A new method for the determination of N,N-dimethyl-5-hydroxytryptamine (bufotenine), N,N-dimethyltryptamine (DMT)*, 5-methoxy-N,N-dimethyltryptamine (5-MeODMT), and N-methyltryptamine (NMT) was developed using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Identification of the analytes is based on liquid chromatographic retention times of analytes and two fragment ions produced by a triple quadrupole mass spectrometer. Quantification is based on electrospray ionization (ESI), and multiple reaction monitoring (MRM) was also utilized for getting better selectivity. The analytes and internal standard were separated from the urine matrix by solid-phase extraction (SPE). The method was applied for the determination of these compounds in urine samples of patients from surgical, medical and psychiatric wards. Of the dimethylated amines, only bufotenine was found in significant amounts (up to 34 microg/L). In keeping with our earlier results, the bufotenine excretion of psychiatric patients was found to be higher than that of the somatic patients. Method, procedure, considerations, statistical evaluations and urine sample spectra are presented.

Ayahoasca: an experimental psychosis that mirrors the transmethylation hypothesis of schizophrenia.

The experimental psychosis observed after drinking Ayahoasca, a South American hallucinogenic beverage from the Amazon Indians, reproduces the pathologic transmethylation theory of schizophrenia. This theory postulates a decrease in the monoamine oxidase (MAO) activity, which results in the accumulation of methylated indolealkylamines, such as bufotenin (5-hydroxy-N,N-dimethyltryptamine), N,N-dimethyltryptamine (DMT) and 5-methoxy-N,N-dimethyltryptamine. These substances are strong hallucinogens as has been previously confirmed experimentally. On the other hand, it is known that Ayahoasca is a beverage usually prepared by boiling two plants, one of them rich in beta-carbolines, which are naturally occurring strong inhibitors of MAO, and the other with high quantities of DMT. This particular combination reproduces what is supposed to occur under pathologic conditions of different psychoses. The effects of Ayahoasca were studied in subjects, assessing urine levels of DMT by gas chromatography-mass spectrometry (GC-MS) before and after the intake of the beverage. The results of this study confirm that the hallucinogenic compounds detected in the healthy subjects' (post-Hoasca, but not before) urine samples are the same as those found in samples from acute psychotic unmedicated patients. The chemical composition of the Ayahoasca beverage, and of the plant material used for its preparation are also reported as well as psychometric and neuroendocrine subject parameters.

A simple and sensitive quantitation of N,N-dimethyltryptamine by gas chromatography with surface ionization detection.

A simple and sensitive method for determination of N,N-dimethyltryptamine (DMT) by gas chromatography (GC) with surface ionization detection (SID) is presented. Whole blood or urine, containing DMT and gramine (internal standard), was subjected to solid-phase extraction with a Sep-Pak C18 cartridge before analysis by GC-SID. The calibration curve was linear in the DMT range of 1.25-20 ng/mL blood or urine. The detection limit of DMT was about 0.5 ng/mL (10 pg on-column). The recovery of both DMT and gramine spiked in biological fluids was above 86%.

Altered consciousness states and endogenous psychoses: a common molecular pathway?

Interest in the role of indolamines in the pathogenesis of psychoses has been renewed in recent years by the development of atypical antipsychotic drugs such as clozapine, olanzapine, and risperidone, which act on serotonin receptors. Discovery of the hallucinogenic compounds called methylated indolealkyalamines (MIAs) (e.g. N,N-dimethylserotonin, or bufotenin, and N,N-dimethyltryptamine, or DMT) led proponents of the transmethylation hypothesis of schizophrenia to theorize that through some inborn error of metabolism, serotonin or tryptamine might undergo the addition of extra methyl radicals, thereby forming MIAs with hallucinogenic properties. Various studies have attempted to detect the excretion of MIAs, especially DMT, in the body fluids of psychotic patients and normal controls. Some of these studies have demonstrated elevated MIA concentrations in psychotic patients, including those with schizophrenia, compared with normal persons, and others have not. A number of variables may account for these contradictory findings. The mechanism whereby the beverage ayahuasca, which is used in certain cure and divination rituals in the Amazon Basin, exerts its hallucinogenic effects may serve as a model to explain the mechanism underlying hallucinogenic symptoms in schizophrenia and may lend support to the transmethylation hypothesis. Certain studies suggest that specific perceptual disturbances manifested by schizophrenic patients could contribute to progressive deterioration and negative symptomatology. All these findings point to the need for further study of the neurophysiology of MIAs and their pathogenetic role in endogenous psychoses.

[Elimination of N,N-dimethyltryptamine by urine]. / Eliminación de N,N-dimetiltriptamina por la orina.

N,N-dimethiltryptamine (DMT) in urine has been quantified on an 83-psychiatric patient sample. Sample covered patients who had sometimes been administered neuroleptic drugs as well as patients with some particular symptomatology associated to psychotic disorders such as hallucinations, delusions, perception disorders, etc. 43.3% (36 cases) evidenced an abnormally high DMT in urine (0.9-13.5 mcg/24 h). Higher values were more frequently found in both schizophrenic patients, and non-schizophrenic patients with either hallucinations, delusions, anorexia or bulimia. Most patients with DMT normal values (< 0.5 mcg/24 h) presented either mental retardation, cerebral atrophy or dysrhythmias. A very good correlation was found between urinary DMT abnormally high values, and patients' improvement after such patients had been treated with neuroleptic drugs.

Eliminación de N, N-dimetiltriptamina por la orina / Elimination of N, N-dimethyltryptamine by urine

Se dosó la N, N-dimetiltriptamina (DMT) en la orina de 83 pacientes psiquiátricos. En el estudio se incluyeron pacientes que habían estado en algún momento en tratamiento con enurolépticos y pacientes con determinada sintomatología asociada a cuadros psicóticos: alucinaciones, delusiones, trastornos de la percepción, etc. En 36 casos (43,3//) se hallaron valores anormalmente altos de DMT urinaria (0,9-13,5 mcg/24 h). Estos valores se encontraron con mayor frecuencia en pacientes esquizofrénicos y en pacientes no esquizofrénicos tanto con alucinaciones o delusiones, como también anorexia o bulimia. La mayoría de los pacientes con valores normales de DMT (<0,5 mcg/24 h) presentaba retardo mental, atrofia cerebral o disritmias. Se encontró un muy buen correlato entre valores elevado de DMT urinária, mejoría clínica y el tratamiento con enurolépticos (AU)

Eliminación de N, N-dimetiltriptamina por la orina / Elimination of N, N-dimethyltryptamine by urine

Se dosó la N, N-dimetiltriptamina (DMT) en la orina de 83 pacientes psiquiátricos. En el estudio se incluyeron pacientes que habían estado en algún momento en tratamiento con enurolépticos y pacientes con determinada sintomatología asociada a cuadros psicóticos: alucinaciones, delusiones, trastornos de la percepción, etc. En 36 casos (43,3//) se hallaron valores anormalmente altos de DMT urinaria (0,9-13,5 mcg/24 h). Estos valores se encontraron con mayor frecuencia en pacientes esquizofrénicos y en pacientes no esquizofrénicos tanto con alucinaciones o delusiones, como también anorexia o bulimia. La mayoría de los pacientes con valores normales de DMT (<0,5 mcg/24 h) presentaba retardo mental, atrofia cerebral o disritmias. Se encontró un muy buen correlato entre valores elevado de DMT urinária, mejoría clínica y el tratamiento con enurolépticos

Gas chromatographic-mass spectroscopic characterisation of the psychotomimetic indolealkylamines and their in vivo metabolites.

The use of liquid chromatography with on-line fluorescence detection has formed the basis for the separation, characterisation and quantitation of a number of metabolites of the psychotomimetic indolealkylamines N,N-dimethyltryptamine and 5-methoxy-N,N-dimethyltryptamine formed both in vitro and in vivo. Verification of the identity of metabolites has previously been facilitated by the combined use of a number of analytical techniques including multidimensional liquid chromatography and stop-flow spectroscopic analysis. We now describe the combination of liquid chromatography with gas chromatography-mass spectrometry for the unequivocal verification of a number of structurally characteristic metabolites of the psychotomimetic indolealkylamines.

The ion-pair extraction, purification, and liquid chromatographic analysis of indolealkylamines in human urine.

A highly efficient ion-pair extraction technique for the isolation of tryptamine, 5-hydroxytryptamine, and their mono- and N,N-dimethylated derivatives from aqueous media is described. The technique has been used to isolate nanogram quantities of both N, N-dimethyltryptamine and 5-hydroxy-N, N-dimethyltryptamine from large volumes of urine. A rapid and efficient normal-phase liquid chromatographic procedure has also been developed for the subsequent purification of indolealkylamines isolated from urine. The methods described have been used in the measurement of the urinary excretion of 5-hydroxy-N, N-dimethyltryptamine. Analyses were performed by liquid chromatography using a cation-exchange column with online fluorescence detection. Further characterization was achieved by stop-flow spectroscopic analysis of the LC eluant.

A longitudinal study of urinary excretion of N,N,-dimethyltryptamine in psychotic patients.

The excretion of N,N,-dimethyltryptamine (DMT) has been measured in longitudinal studies of five patients with schizophrenic illnesses and in four patients with rapidly or slowly cycling manic-depressive illness. The excretion of DMT was frequently raised in patients when they were psychotic but was usually normal when they had recovered. However, rapid changes in the severity of illness or sudden switches from one mood state to another were not accompanied by changes in the excretion of DMT. These findings contrast with the immediate hallucinogenic effects of an injection of DMT, and suggest that the extracerebral production of DMT (as measured by its urinary excretion) does not provoke the experience of hallucinations in psychotic patients.