ABSTRACT Background. Peganum Harmala (Syrian rue, Nitrariaceae) has a long and widespread history of ethnopharmacological use. Due to its rich and diverse alkaloid content, the traditional application of whole plant material or full alkaloid extract is inherently unspecific and therefore susceptible to side effects. Simultaneously, incorrect phytochemical procedures for its use in ayahuasca-analogues cause intoxications in misinformed users. Aim of the study. Providing harm-reduction by developing easily applicable, safe and effective isolation protocols for harmine, harmaline (dihydroharmine) and tetrahydroharmine (leptaflorine) from Peganum Harmala seed. Materials and methods. Only commonly available equipment and reagents were used for isolating the alkaloids. Following extraction, harmine and harmaline were separated using 2 low-tech methods (selective precipitation using sodium bicarbonate or pH-metry). Then, zinc-acetic acid reduction of both harmine and harmaline was attempted. The identity and purity of the obtained alkaloids were confirmed using microscopy and melting point determination. Results. Using pH-metry for the separation of harmine and harmaline proved rapid, effective and recovered 91% of alkaloids. Selective precipitation of the alkaloids with sodium bicarbonate had a yield of 76%. Hydrogenation of harmaline to tetrahydroharmine using zinc-acetic acid had a yield of 83%. Harmine however could not be hydrogenated by zinc-acetic acid. The melting ranges of the obtained alkaloids were narrow and consistent with literature. Based on these results, isolation protocols were developed. Conclusions. Harmine, harmaline and tetrahydroharmine can be isolated with good yields from Peganum Harmala seed following protocols that are applicable in traditional and other low-tech settings. This study showed zinc-acetic acid to be effective in the reductive synthesis of tetrahydroharmine and also proposes a rapid, precise and high yielding method for the bulk separation of harmine and harmaline using basic pH-metry.

{"references": ["Asgarpanah J, Ramezanloo F, 2012. Chemistry, pharmacology and medicinal properties of Peganum harmala L. African Journal of Pharmacy and Pharmacology Vol. 6(22), pp. 1573-1580", "Callaway JC, 2005. Various alkaloid profiles in decoctions of Banisteriopsis caapi. J Psychoactive Drugs. 2005 Jun;37(2):151-5", "Dos Santos RG, Os\u00f3rio FL, Crippa JAS, Riba J, Zuardi AW, Hallak JEC, 2016a. Antidepressive, anxiolytic, and antiaddictive effects of ayahuasca, psilocybin and lysergic acid diethylamide (LSD): a systematic review of clinical trials published in the last 25 years. Ther Adv Psychopharmacol. 2016 Jun; 6(3): 193\u2013213.", "Dos Santos RG, Balthazar FM, Bouso JC, Hallak JE, 2016b. The current state of research on ayahuasca: A systematic review of human studies assessing psychiatric symptoms, neuropsychological functioning, and neuroimaging. J Psychopharmacol. 2016 Jun 10", "Hasenfratz V, 1927. Harmaline and harmine. Ann. Chim., 1927, [x], 7, 151\u2014 226", "Moloudizargari M, Mikaili P, Aghajanshakeri S, Asghari MH, Shayegh J, 2013. Pharmacological and \ttherapeutic effects of Peganum harmala and its main alkaloids. Pharmacogn Rev. 2013 Jul;7(14):199-212. doi: 10.4103/0973-7847.120524", "Nunes AA, Dos Santos RG, Os\u00f3rio FL, Sanches RF, Crippa JA, Hallak JE, 2016. Effects of Ayahuasca and its Alkaloids on Drug Dependence: A Systematic Literature Review of Quantitative Studies in Animals and Humans. Rev Bras Psiquiatr. 2016 Mar;38(1):65-72. doi: 10.1590/1516-4446-2015-1701", "Shulgin A, Shulgin S, 1997. Tihkal, The Continuation. Edn 1, Transform Press, Berkeley, California 1997"]}