plants Article Biodiversity of β-Carboline Profile of Banisteriopsis caapi and Ayahuasca, a Plant and a Brew with Neuropharmacological Potential Beatriz Werneck Lopes Santos 1 , Regina Célia de Oliveira 2 , Julia Sonsin-Oliveira 2 , Christopher William Fagg 3 , José Beethoven Figueiredo Barbosa 4,5 and Eloisa Dutra Caldas 1,* 1 Laboratory of Toxicology, Department of Pharmacy, Faculty of Health Sciences, University of Brasilia, Brasilia-DF 70910-900, Brazil; [email protected] 2 Department of Botany, Institute of Biology, University of Brasilia, Brasilia-DF 70910-900, Brazil; [email protected] (R.C.d.O.); [email protected] (J.S.-O.) 3 Faculty of Ceilândia, University of Brasilia, Brasilia-DF 72220-275, Brazil; [email protected] 4 Department of Phytotechnology, Federal University of Roraima, Boa Vista-Roraima 69310-000, Brazil; [email protected] 5 Centro Espírita Beneficente União do Vegetal (UDV), Brasília-DF 70040-904, Brazil * Correspondence: [email protected] Received: 29 May 2020; Accepted: 20 June 2020; Published: 9 July 2020 Abstract: Ayahuasca is a psychoactive infusion with a large pharmacological application normally prepared with Banisteriopsis caapi, which contains the monoamine oxidase inhibitors β-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 β-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 β-carbolines present in B. caapi. This biodiversity is a challenge for standardization of the material used in ethnopharmacological studies of B. caapi and ayahuasca. Keywords: ß-carbolines; DMT; Banisteriopsis caapi; ayahuasca; Brazil 1. Introduction Ayahuasca is a psychotropic beverage traditionally used in religious rituals and as medicine among indigenous communities from the Amazonian region, a use that has expanded since the 1970s to all Brazilian regions and more recently to other countries [1,2]. The beverage is usually prepared by the decoction of the liana Banisteriopsis caapi (Spruce ex Griseb.) C.V. Morton and Psychotria viridis Ruiz & Pav [3,4] and may be called by other names, such as vegetal and Daime [1]. Plants 2020, 9, 870; doi:10.3390/plants9070870 www.mdpi.com/journal/plants Plants 2020, 9, 870 2 of 14 Banisteriopsis caapi contains β-carboline alkaloids, mainly harmine, harmaline, and tetrahydroharminePlants 2020, 9, x FOR (THH) PEER REVIEW (Figure 1), which are inhibitors of monoamine oxidases (MAO)2 of 15 [ 5,6]. Psychotria viridisBanisteriopsishas the hallucinogenic caapi containsindole β-carboline N,N-dimethyltryptamine alkaloids, mainly harmine, (DMT; harmaline, Figure1), and a serotonin agonist thattetrahydroharmine causes intense (THH) cognitive (Figure and 1), perceptive which are in modificationshibitors of monoamine when oxidases administered (MAO) [5,6]. parenterally in humansPsychotria [7]. When viridis taken has the orally, hallucinogenic DMT isindole oxidized N,N-dimethyltryptamine by MAO, a process (DMT; Figure that is1), preventeda serotonin by the presence ofagonistβ-carbolines that causes after intense ayahuasca cognitive and ingestion, perceptive allowing modifications the when DMT administered to reach theparenterally serotoninergic in humans [7]. When taken orally, DMT is oxidized by MAO, a process that is prevented by the receptors inpresence the nervous of β-carbolines system after causing ayahuasca the psychotropicingestion, allowing eff ectsthe DMT of the to reach beverage the serotoninergic [4,5,8,9]. Harmine and harmalinereceptors arealso in the shown nervous to system be hallucinogenic causing the psychotropic in humans, effects probably of the beverage due to[4,5,8,9]. their interactionHarmine with serotonin, dopamine,and harmaline benzodiazepine, are also shown to be imidazoline, hallucinogenic andin humans, opiate probably receptors, due to acting their interaction as either with agonists or antagonistsserotonin, [10–12]. dopamine, The activity benzodiazepine, of THH in imidazoline, ayahuasca and is apparently opiate receptors, more acting a function as either ofagonists its inhibition or of antagonists [10–12]. The activity of THH in ayahuasca is apparently more a function of its inhibition serotonin uptakeof serotonin than uptake of its than action of its as action a MAO as a MAO inhibitor inhibitor [13 [13].]. N H3C H3C N CH3 O CH H N 3 Harmine N H3C N CH3 O H Harmaline N NH H N,N dimethyltryptamine (DMT) H3C N CH3 O H Tetrahydroharmine (THH) Figure 1. ChemicalFigure 1. Chemical structures structures of of the the β-carbolines-carbolines harmine, harmine, harmaline, harmaline, and tetrahydroharmine and tetrahydroharmine (THH) present in Banisteriopsis caapi and ayahuasca and of N,N-dimethyltryptamine (DMT), present in Banisteriopsis caapi (THH) presentayahuasca. in and ayahuasca and of N,N-dimethyltryptamine (DMT), present in ayahuasca. The therapeutic potential of ayahuasca, B. caapi, and its components, mainly harmine, was The therapeuticwidely explored potential in the last of ayahuasca,20 years [14]. TheB. caapi β-carbolines, and its present components, in B. caapi were mainly shown harmine, to stimulate was widely explored inneurogenesis the last 20 in yearsvitro and [14 the]. modulation The β-carbolines of brain plasticity present could in contributeB. caapi wereto the antidepressant shown to stimulate neurogenesiseffectsin vitroof ayahuascaand the modulation[15] and harmine of brain [16,17]. plasticity B. caapi could extracts contribute showed to theact antidepressantagainst neurodegenerative diseases, such as Parkinson’s disease [18–21]. Furthermore, various studies have effects of ayahuascainvestigated [15 the] and therapeutic harmine potential [16,17 ].of B.ayahuasca caapi extracts for depression showed and to addiction act against in experimental neurodegenerative diseases, suchanimals as Parkinson’s[22–25] and humans disease [26–28]. [18 –21]. Furthermore, various studies have investigated the therapeutic potentialThe species of ayahuasca B. caapi is an forAmazon depression native giant and liana addiction from the Malpighiaceae in experimental family animals [4,29,30], and [22 –25] and humans [26ayahuasca–28]. brewers recognize different “types” of liana. Langdon [31] has documented at least 18 types of B. caapi by the Siona Indigenous people, highlighting features such as length and width of The speciesthe stem,B. size caapi andis shape an Amazon of the leav nativees, and the giant presence liana or from absence the of Malpighiaceae flowers. Schultes et family al. [32] [4,29,30], and ayahuascaremarked brewers that Amazonian recognize natives different acknowledged “types” at of least liana. 30 different Langdon types [of31 B.] caapi, has documentedwhich all have at least 18 types of differentB. caapi usesby thein their Siona traditional Indigenous medicine. people, Peruvian highlighting brewers (ayahuasquer featuresos) such claim asto distinguish length and width of the stem,as size many and as 10 shape types of B. the caapi leaves, lianas [4]. and the presence or absence of flowers. Schultes et al. [32] remarked that Amazonian natives acknowledged at least 30 different types of B. caapi, which all have different uses in their traditional medicine. Peruvian brewers (ayahuasqueros) claim to distinguish as many as 10 types of B. caapi lianas [4]. Some studies have investigated the phytochemistry of the species used in the preparation of the ayahuasca beverage; however, only few samples were analyzed [4,21,32–34], which limits the possibility to capture the diversity of the chemical profile of the plants. The aim of this study was to make Plants 2020, 9, x FOR PEER REVIEW 3 of 15 Plants 2020, 9, 870 3 of 14 Some studies have investigated the phytochemistry of the species used in the preparation of the ayahuasca beverage; however, only few samples were analyzed [4,21,32–34], which limits the a thoroughpossibility chemical to capture investigation the diversity of the chemicalβ-carbolines profile harmine, of the plan harmaline,ts. The aim and of tetrahydroharminethis study was to levelsmake present a thorough in B. caapi chemicaland in ayahuascainvestigation brew of in samplesthe β-carbolines collected fromharmine, various harmaline, regions in and Brazil. Additionally,tetrahydroharmine other plants levels usedpresent in in the B. preparationcaapi and in ayahuasca of the brew brew were