Author Topic: n,n-dialkylated tryptamines from tryptophol made with yeast? (Read 217 times)

carl_nnabis · « **on:** June 13, 2012, 12:38:58 AM »

Hi!
What about this one, isnt it some sort of otc-tryptamines for everybody?
Tryptophol is made from living yeast cultures using tryptophan decarboxylase, following reduction of the so received tryptamine, this was demonstrated many years ago first. Even a member here had success albeit he did some mistakes during recovering the tryptophol!
http://127.0.0.1/talk/index.php/topic,680.msg12756.html#msg12756
Done right one can obtain nearly a quantitative yield, i have yet to find the original paper belongig to ehrlich in english, but it should be really as simple as about 1g tryptophan per liter, sugar and yeast, ferment one week and evaporate water preferably under vacuum or maybe low heat wont hurt there, then extraction of your obtained tryptophol.
Next one of my favorites, the use of a alkyl/acyl sulfonic acid, i.e. tosyl or mesyl group, theres even a youtube video about tosylation of isobutanol out there!
Then replacement with dialkylamine of choice, i have read on SM dipropylamine is somehow obtainable OTC, dimethylamine can be made using OTC precursor...
What do you think? Looks except for tosylchloride like a very convenient way OTC for dialkyltryptamines?
For our reference guru, the DOI for the original 100 year old paper about fermentation of tryptophan is:
DOI: 10.1002/cber.191204501134

atara · « **Reply #1 on:** June 13, 2012, 01:30:30 AM »

Tosyl chloride is a bitch.

However, the good folks over at sciencemadness worked out a preparation for esityl chloride, that is, ethanesulfonyl chloride, which proceeds from ethyl bromide and sodium thiosulfate with the addition of chlorine:

In a one liter RBF:
EtBr : 0.5 mole MW 109 d 1.46 --> 55 g = 38 ml (I add 40ml)
Sodium thiosulfate.5 H20 : 0.5 mole MW 248 --> 124 g dissolved in 400 ml H20
Acetone 250 ml

After one day of stirring a clear homogeneous solution is obtained. Acetone is then evaporated. According to US2293971 there is added 200 ml of GAA to the 300 ml of solution obtained. Meanwhile in another 1L RBF there is placed 300 g sodium dichloroisocyanurate (MW 220, 85%, 1160 mmole, 55% chlorine -> 140 g active chlorine (2 moles)) covered with a small amount of water. With the help of a teflon tubing through septums bridging the two 1L RBF and going into the solution with the bunte salt chlorine is generated for two hours and passed in by dropping at 0-15°C 250 ml of 32% HCl (2.5 mole) on the DCCA. Be cautious of back pressure jump and frothing! Near the end of generation of the chlorine the solution with bunte salt took a permanent yellow-green tinge. It means excess chlorine was present. After ten minutes it was still yellow and workup was started. The solution is pourred in one liter of water which precipitated 30 g of ethane sulfonyl chloride. Four extractions with 4x50 ml CH2Cl2 were done and all fractions are combined then washed once with sulfite and two times with water. The CH2Cl2 evaporated in vacuo and the yellow and fluid residue is weighted : 62 g is obtained MW 128 -> 468 mmol (94%).

A 94% yield is extraordinary for such a nasty compound... and esityl chloride can activate all sorts of alcohols, including sesamol...

carl_nnabis · « **Reply #2 on:** June 13, 2012, 01:41:51 AM »

Good to know 10g are like 1€ only, every chemical supplier who may is willing to deal with private persons will sell it I think? Its really unsuspicious.
Sesamol? wow nice one, that can be obtained on hydrolysis of paroxetin mentioned as precursor from fresh few days ago. i wonder how its concentration is in natural sources?

lugh · « **Reply #3 on:** June 13, 2012, 02:23:54 AM »

The article by Jackson from the Journal of Biological Chemistry that was mentioned in that thread as well as the one by the same author cited are attached

Two other articles from Rhodium are also attached, one in Russian which was partially translated by Antoncho:

"...Usually the transition from alcohols to amines is realized through
replacement of the -OH group w/haloid or tosylate. The drawbacks of the
said method include: the need for a large xcess of amine, the possibility
of quaternary salt formation and generally low yields of the desired
product...

We found that secondary amines are conveniently alkylated w/phenethyl-,
phenylpropyl- and indolylethyl- alcohols by boiling the equimolar
quantities of the said reragents in benzene or xylene for 4 hours. The
yields of tertiary amines comprise 83-87%.
... By a known method (through bromide or tosylate),
2-N,N-trimethyltryptamine was obtained at overall yield of 46,5 %,
5-ethoxytryptamine - 30%. Using our method the above amines were prepared
from the corresponding tryptophols in 81-98% yield.
An additional advantage is that this rxn doesn't require inert atmosphere,
protection from moisture and is generally non-demanding."

A typical synthetic procedure:
[see Rhodium's post]

The yields: DMT (in benzene) - 93%, mp as hydrochloride 221-222 C;
2,N,N-TMT (xylene) - 89%, mp 231-232 C; 2-methyl-DET(xylene) - 81.5%, mp
206-207 C; morpholyltryptamine(xylene) - 91,3%, mp 211-212 C;
2-methylmorpholyltryptamine(xylene) - 98%, mp 248-249 C.
The yields for phenethyl- and phenylpropyl alcohols are 83-86%,
w/morpholine and piperidine as the amines.

        Rhodium
(Chief Bee)
05-26-00 13:04
No 9737
                  Re: maybe 4 steps from tryptophan to DMT/DET/DPT
(Rated as: excellent)

This is the article from J Org Chem USSR, 5, 2158 (1969) mentioned above

General procedure for the preparation of dialkyltryptamines:

Into a flask fitted with a Dean-Stark recieving trap were placed 0.1 moles of tryptophol [2-(3-indolyl)-ethanol], 0.1 moles of a secondary amine [dimethylamine for DMT], 4ml of skeletal nickel suspension in water and 100ml of benzene or xylene [I assume toluene is good too]. The reaction mixture was refluxed, and the condensed water collected for four hours, the hot solution filtered and the catalyst washed repeatedly with benzene or xylene [or toluene] and the mother-liquor concentrated under vacuum. The product was either distilled in vacuo or isolated as the hydrochloride by acidifying with an equivalent of hydrogen chloride solution [they are probably referring to a non-aqueous HCl solution here]. The yields were in the 82-98% range.

Preparation of skeletal nickle: http://water-cooler.com/WC/patentviewer/patent-4086264.html
Perhaps one could use RaneyNi or just powdered/precipitated nickle?

and from:

http://parazite.nn.fi/hiveboard/tryptamine/000009775.html

in_outsider's post and psyloxy's translation from Erlich's article:

        in_outsider
(Stranger)
05-26-00 23:31
No 9775
                  Tryptophol from Tryptophan via yeast

After reading the other Tryptophan thread, I remembered this article. Seems simple enough in theory. Any bees ever come across further references to processes similar to this?

---

Ehrlich, F., Ber. chem. Ges,. 45, 883 (1912).

Von den Gärprodukten der lebenden Hefe, deren Entstehung aus Aminosäuren sich auf Grund der früher <1> aufgestellten Gleichung

R.CH(NH2).CO2H + H2O == R.CH2OH + CO2 + NH3

voraussetzen ließ, verdienen die Alkohole aus Aminosäuren der aromatischen und heterocyclischen Reihe besondere Beachtung, da es sich hier meist um bisher unbekannte, sehr eigenartige chemisch Verbindungen handelt. In dieser Hinsicht reiht sich dem vor einiger Zeit beschriebenon Tyrosol aus Tyrosin der jetzt neuaufgefundene B-Indolyl-äthylalkohol ebenbürtig an, dem ich wegen seiner analogen Beziehungen zum Tryptophan den Namen "Tryptophol" geben möchte, und dessen Bildung zufolge der nachstehenden Formelbilder ohne weiteres verständlich ist:

------

Tryptophol entsteht ähnlich wie Tyrosol, wenn man Hefe auf Lösungen von Tryptophan mit dem üblichen Zusatz von Zucker und anorganischen Nährsalzen wachsen läßt, oder man Tryptophan direkt mit viel Zucker und Preßhefe vergärt. Für die präparative Bereitung des neuen Alkohols ist das letztere Verfahren vorzuziehen, da es in kurzer Zeit unter günstigen Bedingungen Ausbeuten bis zu 80% der Theorie an reinem Tryptophol liefert:

5g tryptophan (aus casein) wurden zusammen mit 600g Zucker Raffinade in 6l Wasser gelöst und die Lösung in einer großen, mit Gärvershluß versehenen Flasche mit 400g frischer Preßhefe Rasse XXI bei Zimmertemperatur vergoren. Nach 3 Tagen ließ sich in der Flüssigkeit kein Zucker mehr nachwiesen. Die von der Hefe durch Tonfilter klar abgesaugte Lösung ergab, im Vakuum bei 40-50 verdampft, einen braunen Sirup, den man mit dem mehrfachen Volumen Alkohol verrührte. Das alkoholische Filtrat hinterließ beim Verdampfen einen sirupösen Rückstand, der, mit 200ccm Wasser aufgenommen und mit 10g festem ätznatron versetzt, einige Minuten auf dem Wasserbade erwärmt wurde. Hierbei trübte sich die Flüssigkeit emulsionsartig, und es schied sich ein dunkelgefärbtes öl ab, das beim Abkühlen bald zu Krystallschuppen erstarrte und leicht in äther überging. Nach erschöpfender Extraktion ergaben die gesammelten ätherauszüge beim Vertreiben des äthers ein bräunliches, eigentümlich riechendes öl, das sich beim Reiben in kurzer Zeit in eine eisblumenähnliche Krystallmasse (3.3 g) verwandelte <2>. Zur Reinigung wurde das Produkt in ca. 3/4l kochendem Wasser gelöst, nach Zusatz von etwas Kieselgur <3> in der Siedeheitze von ungelösten öligen Verunreinigungen durch Filtration befreit und das gelbliche Filtrat auf ein kleines Volumen eingeengt. Die beim Abkühlen der Lösung entstehende Emulsion geht spontan oder durch Impfen und Reiben nach einiger Zeit in ein Haufwerk glänzender Krystallflitter über. Die vollständige Reinigung von geringen Mengen hartnäckig anhaftenden öls gelingt, wenn man die so vorbehandelte Substanz in wenig äther löst, die Lösungmit Tierkohle schüttelt, filtriert, aus dem Filtrat zunächst mit wenig Petroläther bräunlich gefärbte Verunreinigungen niederschlägt und dann die abdekantierte, farblose Flüssigkeit mit Petroläther übersättigt. Aus der stark getrübten Lösung krystallisiert das Tryptophol allmählich in Form von büschelartig angeordneten, weißen, glänzenden Nädelchem und Blättchem, die, im Vakuum über Schwefelsäure getrocknet, scharf bei 59 zu einer farblosen Flüssigkeit schmelzen.

0.1300 g Sbst.: 0.3542g CO2, 0.0820 g H2O - 0.1231 g Sbst.: 9.7ccm N (22 degrees, 752mm).

C10 H11 O N

Ber. C 74.53, H 6.83, N 8.70
Gef. C 74.31, H 7.06, N 9.02

<1> F. Ehrlich, B. 40, 1047 (1907), 44, 139 (1911)

<2> Da Tyrosol aus ätzalkalisher Lösung nicht mit äther zu extrahieren ist, so bietet das beschriebene Verfahren auch einen einfachen Weg, um Tryptophol neben Tyrosol in Gärprodukten nachzuweisen und daraus zu isolieren.

<3> Noch besser gelingt die Reinigung durch Aufkochem der wäßrigen Lösung mit Tierkohle, doch tritt hierbei ein erheblicher Verlust an Substanz infolge Adsorption durch die Kohle ein.



        psyloxy
(Hive Bee)
05-28-00 22:11
No 10417
                  Re: Tryptophol from Tryptophan via yeast
This is my 'a bit less hacked together' variation. I stopped when the clean up part begins, I'll post it later if I find the time, maybe someone else can check what I've written, I'm not completely sure if I got everything right.

---

Tryptophol is produced if one grows yeast in solutions of tryptophan with the usual addition of sugar and anorganic salts or if one directly ferments tryptophan with a lot of sugar and preßhefe.[no idea what preßhefe is, looks like some special sort of yeast, maybe they only mean a concentrated form...]

For tryptophol the latter procedure is preferred because it is faster, works with better conditions and gives yields up to 80%:

5g tryptophan and 600g zuckerraffinade [this is nohing uncommon, something like 'refined sugar', just sugar only very clean] are dissolved in 6000ml H2O and the solution is fermentet at room temp in a big flask with a gärverschluß [I think this is a valve that can stand the pressure, visit your brewery store and you'll see such a thing] with 400g of fresh preßhefe (yeast).

After 3 days there was no sugar left in the liquid. The yeast was filtered out with a clay filter(and vacuum?), the water was evaporated under vacuum at 40-50°C and a brown syrup was left. The syrup was mixed with alcohol(multiple volume). The alcoholic filtrate was evaprated and left a syrupy residue which was taken up in 200ml H2O and mixed with 10g solid NaOH, followed by heating on a water bath for some minutes. During this time the liquid became cloudy like an emulsion and a dark oil separated. After the solution had cooled, the oil crystallized and the crystals were easily taken up in ether. The product of the evaporation of the combined ether extracts was a browny, strange smelling oil that,during friction, quickly turned into a crystalline mass weighting 3.3g.

The product was dissolved in 750ml of boiling H2O...

and a post claiming success:

5 or 6g tryptophol was dissolved in approx 6L warm water then sugar added according to the turbo yeast instructions for fermenting 18% ethanol. after cooling to room temp turbo yeast was added, a gas trap put in place, and left to ferment for 5 days.

solution is filtered then concentrated to a dark residue on the water bath at aspirator vacuum. the still warm residue is dissolved in 200ml 99% IPA (swim doesn't remember, use more IPA if necessary) with heating and filtered again. the IPA is stripped off under vacuum on a water bath and the residue taken up in 200ml 10% NaOH.

the warm (not higher than 40 C) basic solution is extracted with 100ml diethyl ether. the ether is chilled to precipitate the tryptophol as white flakes which are filtered. the ether can then be reused to extract the aqueous layer again.

swim's yield was only .7g but he accidentally extracted first with petroleum ether and it probably held a lot product but would not precipitate it. he did not strip off the petroleum ether so his total yield is unknown. the tryptophol is white crystals melting at 57C. it is a little greyish but still sparkles after a year or two of storage at STP.

the only hassle is stripping all the h2o under vacuum. we need to determine how if the solvent/yeast volume can be reduced while maintaining good yields and this could be a perfectly viable method for ounce-level production if MHRB goes dry. perhaps agitation or use of PTC could reduce the amount of yeast needed.

as Shulgin noted, there's a lot of published information on the subject

carl_nnabis · « **Reply #4 on:** June 15, 2012, 01:43:54 AM »

thank you lugh!

i knew somewhere was that whole idea floating around...^^ but forgot where, yes in the old files of course

translation for the workup wanted`anyone?

JustDreaming · « **Reply #5 on:** June 16, 2012, 05:52:11 PM »

Activated carbon has been seen in small scale tests to work in a S.P.E. to collect tryptophol from a fermentation(added NaCl to increase ionic nature of the solution and increase the hydrophobic attraction with the solid phase). The problem with this is recovery only minute amounts can be recovered(100mg or so from a 1g tryptophan fermentation). Oxidation could theoretically also occur. Maybe an ultrasonic bath for desorption would be beneficial. Less can be more in this instance, over doing the solid phase will make recovery impossible.

Does anyone know if yeast metabolites adhere to the cell walls? If so what are some idea's for cellular lysis? Freezing, ultrasound, blending?

100mL of diethyl ether does not sound like enough for pulls(in the one example). In the original paper they use something like 3 liters of ethyl acetate for a liter of fermentation. Adding solvent to these fermantations also creates pretty yucky micro emulsions.Where yeast cells migrate into the solvent layer...

The idea of using turboyeast to my limited understanding is unnecessary. The fermentation should only run for 2-3 days after the substrate has been added after this point yields will diminish.

See no reason why TsCl is a 'bitch', it seems a very valuable reagent to have at hand. A similar synthetic methodology is used in the preparation of similar pharmacuetical compounds(with MES iirc).

Think a synthetic reduction method might be more practical than fermentation. Unless a solid phase extraction can be worked out.

carl_nnabis · « **Reply #6 on:** June 24, 2012, 10:18:25 AM »

Quote

Think a synthetic reduction method might be more practical than fermentation. Unless a solid phase extraction can be worked out.

I am still very undecided $:-\$
of course its a big problem to recover ones product in a synthesis involving living cells due to the volume to reagent ratio as most of these are toxic for the yeast.
The starting amount can surely bee tweaked higher as tryptophan is not that toxic for yeast i think? its for sure less toxic as it is benzaldehyde, and supernarket yeast is known to stand a few grams of that well.
Also we dont need to keep the heat stabile relative high boiling tryptophol from racemising as chirality gets lost after amination. So it wouldt ne a big problem to simply simmer the filtered soup at 7-80°C and i read somewhere that is enough that on cooling and standing raw and very impure tryptophol already crystallizes...

Reduction of O-indoleacetic acid is rather impractical for me, as in europe the starting material is mostly not in use anymore (EU-laws and such

). and if its avaible then its unfortunately pretty expensive even for very small amounts

But one has to pay attention not to reduce anything in the indole nucleus of course...
im open-minded for every other convenient reduction method, but can neither imagine nor find a good one, but i have to confess i have not searched that hard

I believe tin and acid would be too harsh to get from NH2 to OH without reducing the indole?

And TsCl is really in my opinion the most harmless organic sulfonic acid chloride i know of, and one of the most common too, MSA-chloride is in opposition really toxic and beside a liquid too! Tosyl chloride is a solid, melting above 60°C IIRC, and its so called "vigorous" hydrolysis with water as it is stated in the literature... has to be catalyzed using heat and alkanine, otherwise it can rest a week without any significant change in water

Unless in solution its not very harmful, eye protection is worn in any case anyway working with such caustic stuff. If your TsCl comes as powder, then wear some dust-filter mask too off course, they are avaible at the hardware store to avoid breathing in of saw-dust.

JustDreaming · « **Reply #7 on:** June 29, 2012, 09:18:50 PM »

Quote

and i read somewhere that is enough that on cooling and standing raw and very impure tryptophol already crystallizes..

Where did you read this? Has it been repeated? That sounds most interesting.

An experimenter left some notes about the solid phase extraction:
A gram of L-Tryp was added to a liter of tomatoe paste based fermentation after a day of going alone, and allowed to ferment for 2-3 more days. The solution was then boiled in a sauce pan on low heat to drive off 1/3 of the the solution, primarily the ethanolic fraction. Non-iodized salt was there added to increase the ionic nature, reduce further fermentation, and increase the attraction towards the solid phase. Distilled water washed then heat treated activated carbon granules(aquarium grade*lab grade would be better* 2.5g) were added to the fermentation broth and put on a magnetic stirrer for 12 hours in a vain attempt to reach equilibrium. The solution was decanted and the granules were air dried.

Using methanol to elute through the solid phase left a fuzzy indolic smelling purple and whitish negligable yielding precipitate. However when the granules were stirred first with a 0.1 M NaOH solution for a few hours, dried then stirred with acetone or methanol. When the solvent was distilled from a boiling water bath, suspected tryptophol and surely contaminates were recovered in small quantities.If the flask were submerged in hot water the off white solid would melt to a yellow liquid. 50-100mg yields of this uncharacterized product were noted with this method.

Maybe some fine tuning could be done here by someone more knowledgeable about solid phase extractions.

carl_nnabis · « **Reply #8 on:** July 01, 2012, 04:53:02 AM »

actually i simplified it too much, i mixed something up in my brain i think

correct i should have said, the filtered solution from the fermentation is reduced under vacuo (at 40-50°C) to a syrup, which is taken up in several times its amount in alkohol, filtered and when this alcoholic solution is reduced in volume and cooled, crystalline tryptophol precipitates.
but anyway this sounds not much more complicated. its taken from the german part about the fermentation, here in this thread.

carl_nnabis · « **Reply #9 on:** August 23, 2012, 05:04:40 AM »

for those of you who dont read both boards, ive already posted it in the collective:

Quote from: carl_nnabis on August 23, 2012, 05:00:18 AM

so i was playing around a bit with yeast and tryptophan, and found out that 12,5g tryptophan are tolerable without any problem in only one liter medium!
only the workup sucked and so I got less than 3g out of it
but pretty pure from melting point so thats ok.
I added an half litre water during course of fermentation, to dilute the amount of alcohol so that they wont kill themselves.

have to wait for some pyridine and then im going to tosylate it, then making DET out of it

very viable way!

Author Topic: n,n-dialkylated tryptamines from tryptophol made with yeast? (Read 217 times)

carl_nnabis

n,n-dialkylated tryptamines from tryptophol made with yeast?

atara

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

carl_nnabis

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

lugh

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

carl_nnabis

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

JustDreaming

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

carl_nnabis

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

JustDreaming

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

carl_nnabis

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?

carl_nnabis

Re: n,n-dialkylated tryptamines from tryptophol made with yeast?