Ayahuasca: alkaloids, plants & analogs
Section 3 : Part 2 :
Reported Chemistry of Mimosa hostilis / Mimosa tenuiflora
0.31% of an alkaloid identified as nigerine was found in its root bark (As Mimosa hostilis; fresh roots obtained 29 October 1942 from Brejo dos Padres). [Nigerine was later shown identical with DMT.]
- Page 76 of Gonçalves de Lima 1946
0.57% DMT, by dry weight, was isolated from "roots" obtained from Prof. Oswaldo Gonçalves de Lima. [See account below; this is a calculated figure]
- Page 1286 of Pachter et al. 1959
(This paper also states that 0.51% alkaloid yield was obtained from root bark by Gonçalves de Lima who was unable to identify the alkaloid. Whether this is a typo or whether Gonçalves de Lima 1946 had a typo or whether there is yet another reason for the discrepancy is not known to me. Similarly, unclear is why Pachter listed "roots" rather than rootbark. Whether this was referable to root bark including the rootlets which are commonly attached or whether Gonçalves de Lima actually sent them intact roots is not known to me.
~ 0.003% DMT, by dry weight, has been isolated from the stem-bark.
0.001% Serotonin, by dry weight, was also recovered.
5-MeO-DMT was not found. (They ran it only as a pure reference sample.)
(Identified as Mimosa tenuifolia; [from Mexico]. The dried bark was toasted at 150°r;C for 2 hours and sliced finely (as was done for medicinal use) prior to its analysis.) [1]
- Meckes-Lozoya et al. 1990
Co-TLC run on commercially obtained root-bark (Mexico) detected four (4) very strong indolic bands: 1 was the same Rf as DMT, 2 were much higher in Rf and one was lower (possibly N-methyltryptamine) . Co-TLC with known reference standard by Appleseed. (Xanthydrol spray used to visualize) Counter-culture rumor indicates this root bark to contain 1% DMT but a quantitative assay has not been performed to confirm or reject this nor has the source of the rumor been tracked down.
- Pachter et al. 1959, Journal of Organic Chemistry 24: 1285-1287:
Using dried Mimosa hostilis roots.
2.7 kg of ground roots was exhaustively extracted with Ethanol.
After removal of alcohol, the residue was stirred with 500 ml of 5% aqueous Ammonia and 2 liters of Chloroform. They used a centrifuge to separate the layers. The resulting emulsion was extracted several times with aqueous Ammonia and Chloroform until it contained no alkaloids.
The combined aqueous extracts were then extracted three additional times with one liter portions of Chloroform.
All Chloroform extracts were combined, concentrated and extracted with 2% Hydrochloric acid until exhausted of alkaloids (They used Mayer's reagent to check this.)
The combined acid fractions were brought to pH 9 with concentrated Ammonium hydroxide and then extracted three times with 500 ml portions of Chloroform.
They used a centrifuge to separate the emulsions.
After drying over Magnesium sulfate the Chloroform fractions were evaporated to dryness yielding 18.5 grams of a crude brown alkaloid.
Part of the crude alkaloid was dissolved in boiling ether and filtered to remove black material.
The Ether was then evaporated to dryness and the residue dissolved in 30 ml of Methanol.
An excess amount of Picric acid was dissolved in 75 ml of Methanol and added to the alkaloid solution to yield a crystalline picrate which separated from the solution. Based on crude alkaloid weight. They used 12 gm of crude alkaloid; adding 15 gm of Picric acid.
[12 grams of crude alkaloid yielded 21.9 grams of picrate of which they recovered a total of 20.7 after three recrystallizations from Hexane. 16.9 grams as first crop and 3.8 as second.]
This was filtered and dried.
After converion to the free base it was crystallized from hexane containing a little ethyl acetate.
10 grams of the picrate yielded 3.3 grams of base.
The DMT content of the roots was reported to be 0.57%; as calculated from the picrate.
If they had used all of their crude base, their end yield would have been 10.53 grams; a final return of 0.39% of highly purified DMT (as calculated from the potential final yield)
Notes #
- This might have reduced their yield somewhat but perhaps another point should be considered; that being the frequent presence of tannins in barks used to treat burns. Lou et al. 1965 felt that bark tannins prevented them from recovering more than a small portion of the bark alkaloids from Acacia confusa by binding them in an alkaloid-tannin precipitate when the solution was basified and extracted. This is due to multiple hydrogen bonding sites on tannin.
It may be found helpful to mix powdered Magnesium oxide or Calcium oxide to the dried and powdered plant material, moisten it (be sure pH is around 9 or above), dry it and then do the initial extraction with a nonpolar organic solvent to eliminate tannins at the first step. Tannins are insoluble in most organic solvents; the less polar the solvent the less soluble the tannins. Tannins extracted into chloroform can be removed by washing the solution with 1% aqueous sodium chloride and then drying it over anhydrous sodium sulfate for a few hours.
Tannins are incompatible with strongly basic solutions and form precipitates. Use of the base breaks alkaloid-acid conjugates to free bases without extracting the alkaloids into an aqueous solution. Pachter's approach is also good for this. [An excellent modification would be that used by Poisson when extracting mescaline from T. pachanoi. Poisson took his crude isolate and vacuum distilled it to yield highly purified alkaloid. DMT readily vacuum distills.]
Lou et al. 1965 reported that when they brought their aqueous extract to pH 9 and extracted with chloroform they found their alkaloids formed an "nearly insoluble precipitate" with the tannins present and they could only extract a minor portion of the bases. [Contrast this with the approach of Pachter and associates on the next page.]
Dried powered bark (6 kg.) was macerated with ethanol for 12 hours and vacuum filtered They repeated until no longer reacted with Dragendorff's. Combined filtrates were passed slowly through Amberlite IR-120-H. After washing column with ethanol they eluted with 4% NaOH in 70% ethanol. After concentration in vacuum the residue extracted into benzene. Crystals formed in the benzene and were recrystallized from chloroform. Final crystallization was from benzene by adding petroleum ether to a concentrated solution until cloudy.
Identified by melting point of base & salts, IR & NMR. They noted other van Urk positive spots were observed in crude alkaloid extract while only one was observed in fresh bark. They attributed this to the sensitivity of the base to air.
An interesting solution to dealing with tannins was reported by the use of an extraction acid at pH 1. This is acidic enough to degrade tannins eliminating their input. We have heard success using hydrochloric acid and also ethanol acidified with supersaturated citric acid.