Lilienthal wrote:
"your product is a mixture of everything soluble in aqueous MeOH / EtOH and unsoluble in ethanol"
True. But the undesired components do not interfere enough with the solubility of the major psychoactive component (PSOP) to make the standardized extract unreliable.
But hey - look at this. It is about the use of 4-DMCA instead of 4-DMBA as main ingredient in Ehrlich's reagent. 4-DMCA is OTC and most likely useful as indicator when a simple piece of typing paper is used as TLC surface.
From the paper then all different components can be separately recognized, -extracted and purified with only alcohol as solvent.
Of course the results will probably be better (less losses)with more difficult to obtain solvents and real TLC paper, but given the fact that the desired alkaloids can readily be biosynthesized from brown rice or malt agar, a crude typing paper TLC with 4-DMCA as OTC color indicator can be the finishing touch.
(newbees who have no idea about TLC and mushrooms may also read the text
http://www.erowid.org/plants/mushrooms/mushrooms_article2.shtml
)
from:
Occurence of 5-hydroxylated indole derivatives in Paneolina foenescii (Fries) Kuehner from various origin. T. Stijve, C. Hischenhuber, D. Ashley. Z. Mycol. 50: 361 (1984)
Ehrlich's reagent (Révélateurs pour la chromatographie en couches minces et sur papier, E. Merck, Darmstadt 1975, no 91, p. 32) was initially used because it yielded brightly coloured spots with most of the compounds of interest. However, detection of psilocybin required a few minutes heating at 100 deg. C and under these circumstances co-extracted urea yielded a brightly yellow zone which interfered with the evaluation of serotonin, a major constituent. In addition, sensitivity for tryptophan was poor.
Better results were obtaines using 4-dimethylamino cinnamaldehyde (DMCA) (Fluka no 39421) which was used as a solution of 0,5 g in 10 ml fuming concentrated hydrochloric acid, mixed with 50 ml methanol. This reagent was more sensitive than its benzaldehyde analogue and did not need heating to react with the various indoles. In addition, it produced a different shade of colour with each compound. For example, psilocin turned greenish gray, psilocybin reddish, bufotenin violet, 5-hydroxyindole acetic acid green, serotonin and 5-hydroxytryptophan bright blue, tryptophan purple and tryptamine purple red.
It should be pointed out that these colours may vary with the chemical nature of the TLC adsorbent. For example, psilocybin spots are reddish on Si02 and on SilCel layers, but violet on cellulose.
The reagent proved tobe remarkably sensitive: the detection limitfor serotonin and 5-hydroxytryptophan was 10 ng and for psilocybin and tryptophan 25 ng. At room temperature psiocybin was the last of the indolic ompounds to become visible. Usually, optimal visibility was obtained after 10-15 mm. The reaction could be accelerated by slightly heating with a stream of warm air from a hair-dryer.
Interestingly, the DMCA reagent reacted only very slowly with urea, which yielded a reddish spot only after a few hours, and thus did not interfere with the determination of serotonin.
