Using TLC to guide discovery of hypothetical new Kanna substitutes Caution: No thorough toxicological evaluation of these plants has been undertaken, aside from the more well known research into S. tortuosum etc. This information is not intended to encourage consumption or use of any of these plants Early initial crude phytochemical screening of diverse Mesembryanthemums narrowed down several as potentially mesembrine-type alkaloid containing and revealed diverse phytochemistry that is largely unexplored. While the first paper was primarily concerned with Trichodiadema spp. and D. bosseranum as potential plants of interest, three novel and mostly unexplored plants were highlighted for further study: Drosanthemum floribundum Pink Champagne Pig Face 1 * Also named Drosanthemum hispidum by some Preliminary phytochemical studies by Smith et al. (1998) [1] revealed the presence of 4'-O- demethylmesembrenol in Drosanthemum hispidum and it was assumed based on a) close botanical similarity and b) the TLC finding of a band at Rf = 0.38 that there may be mesembrine analogues present, so it was selected as a promising lead. On the contrary, Gaffney (2006) screened diverse succulents via TLC and GC-MS and found several Drosanthemum spp. to test negative to alkaloids via TLC. While largely unexplored, the 4'-O-demethylated mesembrine-type alkaloids are found in Sceletium emarcidum , a plant that is culturally used as a Kougoed product and still reputedly retains some desired efficacy. In silico analyses have noted potential GABA-A, AChE and 5- HT affinities from the 4'-O-demethylated constituents [2] A new sample using basified (aq. ammonia) isopropanol was prepared to allow a more concentrated sample of this plant. TLC revealed once again a dense spectrum with confirmation of earlier findings of a major constituent ~ Rf = 0.37 which was noted in the previous paper and hypothesised to (?) potentially be a 4'-O-demethylated mesembrine- type alkaloid constituent, based on similarity to the known mesembrine-containing plants. This was selected as the primarily most interesting plant to pursue for research and potential future bioassays. 2 TLC of Drosanthemum floribundum (Silica, 0.2mm, glass backed, acetone elution, I 2 visualisation) Unidentified Mesembryanthemums selected as leads Also, an unknown red Mesembryanthemum, tentatively identified as a Lampranthus or Malephora spp. showed a rich phytochemical composition in one early TLC and while none corresponded with the hypothesised mesembrine band, it was selected as a potentially interesting plant to explore in further depth. Samples from previous analysis were further concentrated and subjected to TLC. A major slightly pigmented band of Rf = 0.61 was noted in the concentrated sample, with significant trailing of lesser Rf constituents. Interestingly, this one sample has consistently shown differences throughout storage of constituent Rf's, suggesting there may be some unstable constituents or other factors complicating analysis. 3 Red Mesembryanthemum TLC (Silica, 0.2mm, glass backed, acetone elution, I 2 visualisation) Some studies of broader Lampranthus or Malephora have suggested tortuosamine-type alkaloids as a potential constituent [4], despite other analyses sometimes finding the entire Ruschioideae section devoid of alkaloids (Including Gaffney 2006) While there seems to be quite rich phytochemical diversity in this red Mesembryanthemum, it was concluded there was not enough evidence to suggest it may be a promising substitute. Another unknown Mesembryanthemum showed extreme similarity to Lampranthus spectabilis in it's phytochemistry on initial TLC and similar growth characteristics, which is currently being used as a Kanna substitute so that was also selected as a promising lead. Revisiting the TLC for this plant gave the following: 4 There was significant trailing of the constituents but notable bands Rf = ~0.33 (broad). 0.8 and visibly pigmented constituents at 0.93. This broad range of phytochemicals with indistinct Rfs as bands is similar to the initial crude TLC results. It was concluded that it was possibly an interesting candidate for bioassay if toxicology could be established. References [1] Smith, Michael T.; Field, Courtney R.; Crouch, Neil R.; Hirst, Manton (1998). "The Distribution of Mesembrine Alkaloids in Selected Taxa of the Mesembryanthemaceae and their Modification in the Sceletium Derived 'Kougoed'". Pharmaceutical Biology. 36 (3): 173–179 https://doi.org/10.1076%2Fphbi.36.3.173.6350 [2] Reddy Kaylan, Stander Marietjie A., Stafford Gary I., Makunga Nokwanda P., Mass Spectrometry Metabolomics and Feature-Based Molecular Networking Reveals Popula Ɵ on Specific Chemistry in Some Species of the Scele Ɵ um Genus (2022) , Fron Ɵ ers in Nutrition, 9, https://doi.org/10.3389%2Ffnut.2022.819753 [3] Candice Delphine Gaffney, A study of Mesembryanthemaceae alkaloids (2006) https://def-sa.com/wp-content/uploads/2020/08/Gaffney_Candice_D_2006_0.pdf [4] Anti-Alzheimer potential, metabolomic profiling and molecular docking of green synthesized silver nanoparticles of Lampranthus coccineus and Malephora lutea aqueous extracts https://doi.org/10.1371/journal.pone.0223781 5