A preliminary TLC study of Bobinsana - Calliandra angustifolia Calliandra angustifolia ( Bobinsana) is a Fabaceae , in the mimosoid clade of the subfamily Caesalpinioideae with traditional use as an anti-rheumatic, contraceptive, tonic, stimulant, and depurative and a decoction of the whole plant has been suggested to be a "general energising tonic", taken for strength and energy [1]. Effects have been stated to be that Bobinsana "Calms, causes reflections, smooth feelings. Capacity to be flexible" [2], the plant teaching "Rooting, affective communication, openness of heart" . More spiritually, "Shipibo traditions revere the Master Plant Teacher for her gentle heart opening, leading to increased compassion, strength, and awareness of/receptivity to spiritual guides" and prescribed in shamanic practices to heal matters of the heart, emotional trauma along with physical ailments [3] While disputed, it is generally believed species of Calliandra do not produce alkaloids, they are characterised instead by the synthesis of non-protein amino acids which are pipecolic acid and their derivatives [4]. Pipecolic acid and 12 different pipecolic acid derivatives have been isolated from C. angustifolia [3,4] In the leaves, the following profile of pipecolic acids was obtained by Romeo, 1984: 1 a) Pipecolic acid b) 5-hydroxypipecolic acid Flavonoids are also prominent with other constituents possibly including (?) alkaloids, amino acids, cyanogenic glycosides, tannins, saponins, and sterols. As exemplified by another leguminous shrub - A. julibrissin - an antidepressant effect, achieved instead through being rich in flavonoids, lignan glucosides, and saponins - means alkaloids are definitely not a prerequisite for CNS activity [5]. While there has been modern mention of MAO inhibitory activity of the plant due to β- carbolines, there is no scientific literature to support these claims "There are some rumors about Bobinsana containing psychoactive alkaloids however these are completely unproven by any lab test and the effects of Bobinsana do not coincide at all with the effects of the rumored alkaloids." Pipecolic acids have been considered a GABAergic modulator that increases the release and decreases the uptake of γ-aminobutyric acid (GABA) by brain neurons [5] Bejar et al., 1995 found pipecolic acid and 5-hydroxypipecolic acid (both isolated in C. angustifolia ) have shown activity due to serotonin (5-HT) antagonism [6]. 2 When studied by Adesina, 1982, a Calliandra spp. has shown CNS depressant activity [7]. Inhibition of COX-1 [8]. Experimental: Bobinsana bark (~5g) was macerated in basified (aq. ammonia) 95% ethanol for several hours, filtered and concentrated to a small sample. TLC was initially tried with acetone giving poor results so a mixed solvent of acetone:white spirits 1:1 gave good resolution of the constituents. Interestingly, UV visualisation gave a strongly fluorescent compound Rf = 0.47 as a major constituent suggesting the possibility of β-carbolines (or other fluorescent constituents) existing, contrary to the current above data suggesting the absence of such. Pipecolic acids etc are not assumed to be fluorescent. Bobinsana bark TLC (acetone:white spirits:1:1) UV visualisation showing the fluorescent band. 3 a) Mixed solvent and b) acetone elution of Bobinsana bark ('Bob') under UV visualisation. The acetone elution gave a strongly fluorescent compound for the Ref sample at Rf = 0.88 4 a) Mixed solvent and b) acetone elution of Bobinsana bark ('Bob') I 2 visualisation I 2 visualisation showed a dense spectrum of constituents not seen in UV in the Ref. To narrow down possibilities, a reference of a common known plant material was used rich in β-carbolines. The flourescent band in the Bobinsana seemed distinct to the reference material hence it a possibility that it is a distinct compound. 5 Conclusion: The potential presence of a strongly fluorescent compound in Bobinsana bark is intriguing and suggests that the presence of β-carbolines should not be totally discounted (or it may be another fluorescent compound) but in contrast to more common β-carbolines, it may possibly be a distinct compound References: [1] The Tropical Plant Database https://rain-tree.com/bobinsana.htm [2] Politi M, Friso F (2018). Amazonian medicinal plants botanical garden of takiwasi center in Peru; a case report of 25 years’ hands-on experience. Horticult Int J. 2(3):68‒70. . https://doi.org/10.15406/hij.2018.02.00028 [2] BOBINSANA : Lucid Dreaming Aid & Shamanic Heart Healer https://animamundiherbals.com/blogs/blog/bobinsana-heart-opener-lucid-dream-herb [2] J. Sanz-Biset, S. Canigueral (2011) Plant use in the medicinal practices known as “strict diets” in Chazuta valley (Peruvian Amazon) Journal of Ethnopharmacology 137 271–288 https://doi.org/10.1016/j.jep.2011.05.021 [3] Romeo (1984) Insecticidal Imino Acids in Leaves of Calliandra. Biochemica/Systernatics end Ecology, Vol. 12, No. 3, pp. 293-297 https://doi.org/10.1016/0305-1978(84)90052-8 [4] Romeo, J.T., Swain, L.A., Bleecker, A.B. (1983) Cis-4-hidroxypipecolic acid and 2,4-cis-4,5- trans-4,5-dihydroxypipecolic acid from Calliandra. Phytochemistry 22, 1615–1617. https://doi.org/10.1016/0031-9422(83)80098-3 [5] Huang B, Wu Y, Li C, Tang Q, Zhang Y. Molecular basis and mechanism of action of Albizia julibrissin in depression treatment and clinical application of its formulae. Chin Herb Med. 2023 Mar 15;15(2):201-213. https://doi.org/10.1016/j.chmed.2022.10.004 [5] Gutierrez MC, Delgado-Coello BA (1989) Influence of pipecolic acid on the release and uptake of [3H] GABA from brain slices of mouse cerebral cortex. Neurochem Res 14:405–408 https://doi.org/10.1007/BF00964852 [6] Bejar, E., Amarquaye, A., Che, C.T., Malone, M.H., Fong, H.H.S., 1995. Constituents of Byrsonima crassifolia and their spasmogenic activity. International Journal of Pharmacognosy 33, 25–32. https://doi.org/10.3109/13880209509088143 [7] Adesina, S.K., 1982. Studies on some plants used as anticonvulsants in Amerindian and African traditional medicine. Fitoterapia 53, 147–162. https://eurekamag.com/research/001/129/001129060.php [8] Dunstan, C.A., Noreen, Y., Serrano, G., Cox, P.A., Perera, P., Bohlin, L., 1997. Evaluation of some Samoan and Peruvian medicinal plants by prostaglandin biosynthesis and rat ear oedema assays. Journal of Ethnopharmacology 57, 35–56. 6 https://doi.org/10.1016/s0378-8741(97)00043-3 7