Quantitation of N,N-Dimethyltryptamine and Harmala Alkaloids in Human Plasma after Oral Dosing with Ayahuasca

Abstract

Harmine, harmaline, tetrahydroharmine (THH), and N,N-dimethyltryptamine (DMT) were quantitated in plasma from 15 healthy male volunteers after the ingestion of ayahuasca, a beverage that has been used for religious purposes in Brazil since pre-Columbian times. A growing awareness of the interest in this ancient shamanistic practice in modern urban cultures and the widespread popular dissemination of the inebriant effects and type and sources of the plant admixtures used to prepare the beverage have provided additional impetus for this study. The three harmala alkaloids were quantitated from protein-precipitated plasma by high-performance liquid chromatography using fluorescence detection. Recovery from blank human plasma was quantitative, and the limit of quantitation (LOQ) was below 2 ng/mL of plasma for each of the harmala alkaloids. Standard concentrations ranged from 10 to 250 ng/mL for harmine and THH and from 1.0 to 25.0 ng/mL for harmaline, respectively. Linearity was observed for harmine, harmaline, and THH within these respective ranges. The highest concentrations of harmala alkaloids in human plasma were found to be 222.3 ng/mL for harmine, 134.5 ng/mL for THH, and 9.4 ng/mL for harmaline. DMT was quantitated by gas chromatography using nitrogen-phosphorus detection after liquid-liquid extraction with diphenhydramine as an internal standard. DMT recovery was quantitative, and the limit of detection and LOQ were 0.5 and 5 ng/mL, respectively. Linearity for DMT was observed from 5 to 1000 ng/mL. The one-step extraction method for DMT and the protein precipitation method for the three harmala alkaloids afford rapid, sensitive, and quantitative analyses of these alkaloids with minimal analyte loss. The analytical methods also may be applicable to other matrices, including whole blood and urine samples and homogenized tissue specimens. These are the first reported observations of DMT and harmala alkaloids in plasma after ritual ingestion of ayahuasca.

Gas chromatographic analysis of dimethyltryptamine and β‐carboline alkaloids in ayahuasca, an amazonian psychoactive plant beverage

“…The synthesis of tetrahydroharmine was performed according to the procedure described by Callaway et al (1996). In summary, harmaline hydrochloride (251 mg, 1.0 mmol) was slowly added to a methanolic sodium borohydride solution (38 mg, 1.0 mmol) at 0°C.…”

“…Peruvian ayahuasca (100 mL) has been shown to contain dimethyltryptamine (60 mg), harmine (467 mg), harmaline (41 mg) and tetrahydroharmine (160 mg) using two-dimensional thin-layer chromatography, high-performance liquid chromatography and gas chromatography-mass spectrometry (McKenna et al, 1984), whereas Brazilian ayahuasca (100 mL) has been shown to contain 24, 170, 20 and 107 mg respectively of the same four alkaloids using gas chromatography and high-performance liquid chromatography (Callaway et al, 1996).…”

“…Usually, dimethyltryptamine is determined by gas chromatography using a nitrogen-phosphorous detector (GC-NPD) and β-carbolines by high-performance liquid chromatography (HPLC) in different procedures (Callaway et al, 1996(Callaway et al, , 1999Yritia et al, 2002). However, Kikura-Hanajiri et al (2005), simultaneously detected some hallucinogenic tryptamines/ β-carbolines, including dimethyltryptamine, harmine and harmaline, using gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-electrospray ionisation-mass spectrometry (LC-ESI-MS).…”

Effects of the South American Psychoactive Beverage <i>Ayahuasca </i>on Regional Brain Electrical Activity in Humans: A Functional Neuroimaging Study Using Low-Resolution Electromagnetic Tomography

“…The freeze-dried homogenized material had been obtained from a 9.6-liter batch of ayahuasca. The DMT content in the lyophilizate had been determined by HPLC, as described by Callaway et al [19], and the ß-carboline constituents were determined following a modification of the method described in that study. The 9.6-liter batch yielded 611 g of freeze-dried powder, containing 8.33 mg of DMT, 14.13 mg of harmine, 0.96 mg of harmaline and 11.36 mg of tetrahydroharmine per gram.…”

Ayahuasca : liane de l'âme, chamanes et soumission chimique

“…La méthode de dosage de Callaway appliquée à 15 volontaires ayant consommé de l'ayahuasca (2 ml/kg contenant harmine 1,7 mg/ml, harmaline 0,2 mg/ml, tétrahydroharmine 1,1 mg/ml et DMT 0,25 mg/ml) au cours d'un rite religieux, a permis de mesurer les concentrations sériques moyennes suivantes : harmine 114,6 ng/ml, harmaline 6,2 ng/ml, tétrahydroharmine 90,8 ng/ml et DMT 15,8 ng/ml (8).…”