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.

Determination of psilocin, bufotenine, LSD and its metabolites in serum, plasma and urine by SPE-LC-MS/MS.

A validated method for the simultaneous determination of psilocin, bufotenine, lysergic acid diethylamide and its metabolites in serum, plasma and urine using liquid chromatography-electrospray ionization/tandem mass spectrometry was developed. During the solid-phase extraction procedure with polymeric mixed-mode cation exchange columns, the unstable analytes were protected by ascorbic acid, drying with nitrogen and exclusion of light. The limits of detection and quantitation for all analytes were low. Recovery was ≥86 % for all analytes and no significant matrix effects were observed. Interday and intraday imprecisions at different concentrations ranged from 1.1 to 8.2 % relative standard deviation, bias was within ±5.3 %. Processed samples were stable in the autosampler for at least 2 days. Furthermore, freeze/thaw and long-term stability were investigated. The method was successfully applied to authentic serum and urine samples.

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.

Elevated urine levels of bufotenine in patients with autistic spectrum disorders and schizophrenia.

OBJECTIVE: Previous studies have suggested that the endogeneous psychotomimetic molecule bufotenine (N-N-dimethyl-5-idroxytryptamine) may play a role in the pathogenesis of severe mental disorders. The potential association of bufotenine with the clinical features of autism and schizophrenia is not entirely understood. In this study, we measured urinary levels of bufotenine in subjects with autistic spectrum disorder (ASD), schizophrenia and healthy comparison subjects free of psychiatric symptoms. We also sought to assess whether urine concentrations of this molecule may be associated with the clinical characteristics of psychiatric patients. DESIGN: Urine bufotenine levels were measured using a high-performance liquid chromatography-mass spectrometry (HPLC-MS) assay in young adults with severe ASD (n=15), patients with schizophrenia (n=15), and healthy control subjects (n=18). The Vineland Adaptive Behavior Scale was used to measure adaptive behaviors in ASD individuals. The Brief Psychiatric Rating Scale (BPRS) was used for patients with schizophrenia. RESULTS: Urine bufotenine levels were significantly higher in ASD subjects (3.30 +/- 0.49 microg/L, p<0.05) and patients with schizophrenia (4.39 +/- 0.43 microg/L, p<0.001) compared with controls (1.53 +/- 0.30 microg/L). Among patients with ASD, there was a significant positive correlation between urine bufotenine and hyperactivity scores on the Vineland Adaptive Behavior Scale (r=0.479, p<0.05). No other associations were detected. CONCLUSIONS: Our results indicate that elevated urine levels of the endogeneous psychotomimetic molecule bufotenine may play a role in ASD and schizophrenia, and can be correlated with hyperactivity scores in autism.

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.

Bufotenine reconsidered as a diagnostic indicator of psychiatric disorders.

We have analyzed products of the serotonin-degradative pathway, in which both N-methylserotonin and bufotenine are formed in urine specimens of products with psychiatric disorders by three-dimensional HPLC with electrochemical detection. Bufotenine was detected in urine from all autistic patients with mental retardation and epilepsy (n = 18) and many autistic patients (32/47) with mental retardation. Bufotenine was detected in the urine of 15 of 18 patients with depression. Thirteen of 15 schizophrenic patients were also positive for bufotenine. N-methylserotonin was also detected in some cases of each disorder. Only two of 200 urine specimens from healthy controls were positive for bufotenine. Thus, the presence and levels of bufotenine might be useful and important markers of some psychiatric disorders.

Urinary excretion of bufotenin (N,N-dimethyl-5-hydroxytryptamine) is increased in suspicious violent offenders: a confirmatory study.

We previously reported that violent offenders with paranoid symptoms or whose violent actions had been directed against family members had higher urinary levels of bufotenin than other violent offenders. In the present study, patients were evaluated with the Karolinska Scales of Personality (KSP), and urinary levels of bufotenin were determined by mass spectrometry. In drug-free patients suspiciousness was positively correlated, and socialization was negatively correlated, with urinary bufotenin excretion. These two personality variables were strongly interdependent. In drug users, bufotenin excretion was correlated positively with social desirability and negatively with irritability, but not with suspiciousness. Bufotenin excretion was not found to be associated with violence toward family members in the present study. The results are in keeping with the earlier finding that violent offenders with paranoid personality traits have higher urinary levels of bufotenin than other violent offenders.

Marcadores biologicos en Psiquiatria / Biological markers in Psychiatry

In order to determine wheter psycopathology is associated with charactheristic neurochemical changes of psychiatric patients, Noradrenergic; Dopaminergic and Serotoninergic urine compounds were quantified in 50 patients (32 females and 18 males) between 20 and 60 years old. They were classified in four groups, according to DSM-IV criteria; in MAJOR DEPRESSION (MD) 30 cases, OBSESSIVE COMPULSIVE DISEASE (OCD) 9 cases, BIPOLAR DEPRESSION (BD)4 cases and SCHIZOPHRENIA (SZ) 7 cases. The following Amine metabolites were determined in 24 hours urine samples Phenylethilamine (PEA); 3-Metho-4-Hidroxy Phenilglycol (MHPG); 5-Hidroxy-indol acetic acid (5HIAA) Homovanilic acid (HVM); Bufotenine (BU); Ometil Bufotenine (OMBU) and 3-5 Metoxy-NN-Dymethyltryptamine (MNNDMT). The results showed a dicrease of Pea levels in 43 per cent of DM; 33 per cent of OCD; 25 per cent of BP and 42 per cent SZ. MOPEG levels were disminished in 53 per cent of DM; 66 per cent of OCD; 25 per cent of BP and 85 per cent of SZ. There was an increase of HVM levels in 10 per cent of DM; 11 per cent of OCD and 25 per cent of BP. There was a dicrease of 5-HIAA levels in 10 per cent of DM while it was increased in 33 per cent and 14 per cent of SZ, BU, OMBU and NNDMT were positive in 71 per cent of SZ; 46 per cent of DM; 55 per cent of OCD and 50 per cent of BP. Our findings are consistent with the hypothesis that PEA; MHPG; 5-HIAA and HVM compounds could be "State markers"of DM and HVM of BP and SZ patients. There is further evidence to support a close interrelationship between the three systems and the urinary excretion of methylates compounds specially in SZ and in DM, OCD and BP patients.

Nialamide, an MAO inhibitor, increases urinary excretion of endogenously produced bufotenin in man.

Nialamide, an MAO inhibitor, was given per os (PO) to a normal man who volunteered in two separate trials (total intake 300 mg and 1000 mg, respectively), and his bufotenin excretion was followed by consecutive urine samples. In both experiments the excretion rose well above the values measured from the same test subject when not taking nialamide (median 0.089 nmol/mmol creatinine, range 0.002-1.78). At its highest, the excretion was 16.5 nmol/mmol creatinine, and the maximum urinary output was 495 nmoles (56 micrograms) in 24 hr. The levels of bufotenin in plasma required for the excretion of the latter amounts are not far from those that produce psychic symptoms in man.

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.

Bufotenine reconsidered.

Bufotenine given intravenously to a medically trained volunteer subject correlated with the appearance of profound perceptual and emotional changes which were of short duration. The compound rapidly disappeared from the blood and metabolites quickly appeared in the urine of the subject.

The presence of free and conjugated bufotenin in normal human urine.

The N,N-dimethylated derivative of serotonin, bufotenin , is excreted into normal human urine as a free amine. Conjugation of bufotenin is, however, possible because of the phenolic hydroxyl group on the molecule. In the present study the urinary excretion of free and conjugated bufotenin of ten healthy, drug free subjects was examined. Acid as well as enzymatic hydrolysis was used to liberate the amine from its conjugate. Quantification was achieved by isotope dilution mass fragmentography. Of total urinary bufotenin a relatively constant amount, 59.9 - 69.0%, was excreted in conjugated form. The conjugate was tentatively identified as a glucuronide.

[Identification of N,N-dimethyl-O-(beta-D-glucopyranuronosyl)-5- hydroxytryptamine as a bufotenine metabolite in the rabbit]. / Identifikatsiia N,N-dimetil-O-(beta-D-gliukopiranuronozil)-5- oksitriptamina kak metabolita bufotenina v organizme krolikov.

A bufotenine metabolite has been isolated from the rabbit urine by the column chromatography on cellulose and preparative paper electrophoresis in acidic buffer. It has the structure of N,N-dimethyl-O-(beta-D-glucopyranuronosyl)-5-hydroxytryptamine as proved by comparison of chromatographic, electrophoretic and NMR data with those for respective synthetic O-glucuronide.

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.

Determination of N,N-dimethylindolealkylamines in plasma, blood and urine extracts by radioimmunoassay and high pressure liquid chromatography.

A sensitive radioimmunoassay for N,N-dimethylindolealkylamine derivatives has been developed. It is possible to detect 200 to 700 femtomoles of 5-hydroxy-N,N-dimethyltryptamine, 5-methoxy-N,N-dimethyltryptamine or N,N-dimethyltryptamine in a given sample. Antibodies were produced in rabbits immunized with a conjugate prepared by reacting 5-hydroxy-N,N-dimethyltryptamine with a diazotized dl-p-amino-phenylalanine bovine serum albumin conjugate. For identification of immunoreactive material high pressure liquid chromatography was used to separate these compounds from each other and from known cross-reacting compounds found in physiological specimens. After chromatography, individual fractions were analyzed by the radioimmunoassay. This combination of high pressure liquid chromatography and radioimmunoassay has permitted the identification and quantification of these compounds in extracts of urine, plasma and whole blood from normal individuals.