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Using DMT in substrate to increase ration of 4HO-DMT to 4HO-NMT


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#41 PistolPete13

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Posted 18 December 2018 - 11:05 PM

I translated a couple of Gartz' patents from German awhile ago, it's not perfect but all the important stuff is correct. There was an article in the Entheogen Review years ago, and a guy named Yachaj said the following quote. And it has been repeated a lot since, I will put the patent up soon but nowhere in the patent does it say brown rice flour. It clearly says grains of rice, there is no specification at all to color, variety or anything like that.;

 

Pollock soon concentrated on cheap and commonly available brown rice grains. He found that brown rice medium was highly selective for P. cubensis (no other mushroom could be fruited on it). Indeed, the selective quality of brown rice protects newbie psilonauts against accidentally growing toxic mushrooms. But there are more reasons to think of brown rice as superb. In the 1980s, German mycologist Dr. Jochen Gartz, went so far as to file a patent (No. 88-09773, Akad.Wiss. DDR) on brown rice after his discovery that this medium supported the cultivation of P. cubensis of unprecedented potency—1% psilocybin/psilocin by dry weight (which almost equals Panaeolus cyanescens), the highest natural potency ever reported of this mushroom.

From 'The Entheogen Review'  vol X. no 4. winter solstice 2001. page 127

 

 

DD254395A1

 

Gartz, Jochen, Dr.rer.nat.,DD

 

Process for obtaining tryptamine derivatives by cultivating higher fungi

 

The invention relates to a process for obtaining tryptamine derivatives psilocybin and psilocin by cultivating the fruiting bodies of higher fungi. The active substances are applicable as model substances in neurobiological research. The fruiting bodies are acheived after inoculation of an autoclaved mixture of rice grains and water with the primary mycelial culture after a few weeks, without further operational steps such as additional preparation of the substrate, use of a special

casing or constantly maintaining the humidity. The recovery of the active ingredients from the dried and powdered fruiting bodies by known methods such as extraction and chromatography.

 

Patent Claims:

  1. A process for obtaining tryptamine derivatives by cultivating fruitung bodies of psilocybin and psilocin containing higher fungi, preferably of the genera Psilocybe, Strokaria, Plutens and Conocybe, distinguished by the fact that moist rice grains are used as a solid culture media, innoculated with primary cultures under sterile conditions and after the fruit body formation, the separation of psilocybin and prilocin known per se, as done by extraction or chromatography.

  2. The method according to claim 1, is characterized in that after the growth of the mycelium through the culture media at a temperature of 20-25 * C, the cultivation of Fruit body takes place unsterilized, and depending on the fungus with or without additional exposure.

  3. Culture media for cultivation of higher fungi, which can be used for the production of tryptamine derivatives, characterized by the composition

    - Rice Grains 20-70%

    - Water 30-80%

    the mixture in an autoclaved state.

 

Field of application of the invention

 

The invention relates to a process for obtaining tryptamine derivatives by artificial cultivation of higher fungi on suitable nutrient media. The substances psilocybin and Psilocin are applicable as model substances in neurobiological research.

 

Characterisation of the known technical solutions

 

The tryptamines psilocybin and psilocin are found naturally occurring in higher fungi and are detectable in different species of the genera Psilocybe, Stropkaria, Plutens, Conocybe and Inocybe.

It is known that the fruiting bodies of various types of fungi of the genera Psilocybe and Stropkaria are artificially cultivated on compost substrates(DE 1087321).

From this the active ingredients can be prepared in their crystal form by means of suitable extraction methods and subsequent chromatographic separation. The method described has several disadvantages. An additional and lengthy process step is the preparation of the corresponding compost substrates. Furthermore, a casing layer for fructification is necessary, fruit body formation often begins only after many weeks. Finally, to achieve fructification, constant humidity levels of the air has to be ensured.

Furthermore, a substrate with rye is described as a main component for fruit body generation (J.Ethnopharm.5 [1982] 287). the disadvantage of this method is likewise, the necessary application of a complicated composite casing layer with specific water-binding capacity while ensuring constant fertilization.

 

Object of the invention

 

The aim of the invention is to develop a simple and inexpensive process for obtaining tryptamine derivatives by culturing fruiting bodies of higher fungi,

in which a high concentration of tryptamine derivatives in the fungal tissue is achieved with simultaneous rapid fructification.

 

Explanation of the nature of the invention

 

The inventions object is to select such culture medium for fructification, which ensure a rapid fruit body formation at the same time high concentration of tryptamine derivatives in the mushroom without a casing and the need for constant humidification.

According to the invention the object is achieved by using a mixture of rice grains and water as a solid medium for fruit body formation. After autoclaving the mixture is sterile and the medium is innoculated at room temperature(20 to 25*C) with suitable primary cultures from agar plates. For the nutrient medium, 20 to 70% rice grains and supplemental amounts of water are used to get to 100%. After 3 to 4 weeks growth of the culture with or without additional exposure, per night mushroom species, the first fruiting bodies appear without use of a casing layer or necessary moistening, fruit will be harvested in the next few weeks, depending on the particular fructification wave. After the complete growth of the medium with mycelium, the fruit body formation can be continued even under unsterile conditions. In this procedure, psilocybin and psilocin are in quantities up to 1% and 0.5%, respectively, calculated on the dry masses of the fruiting bodies. The active compounds are prepared in a known manner by extraction with methanol and the following isolated with chromatography.

 

The invention is explained in more detail by the following examples:

 

Embodiments

 

Example 1

 

A primary culture of Psilocybe cubensis (Earle)Singer was used for inoculation which was obtained by germination of the spores on 100ml of 6% malt agar, 14 days after innoculation and subsequent cultivation (20 days) on 100ml of the same agar.

A piece of mycelium about 2 x 2cm was removed from it with a sterile spatula and by shaking with 30ml of sterile water and 15g saddle packing, generated a suspension of fine mycelium flakes.

10ml of the suspension was used to inoculate an autoclaved mixture of 100g rice grains and 200ml of water. After 3 weeks the formation of the fruiting bodies began, which in 5 fructification waves appeared.

 

Yield: 21g dry fruiting bodies

HPLC analysis: Psilocybin content: 0.7%

Psilocin content: 0.1%

 

Workup is carried out in a known manner by extraction of the powdered material with methanol and subsequent column chromatography on cellulose.

 

Example 2

 

A second strain of Psilocybe cubensis (Earle)Singer was used. Under analogous conditions, the first fruiting bodies were reached after 4 weeks.

 

Yield: 25g dry fruiting bodies

HPLC: Psilocybin-content: 0.95%

Psilocin-content: 0.2%



#42 PistolPete13

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Posted 18 December 2018 - 11:17 PM

Now we can start getting to his interesting ones more pertinent to this discussion, this next patent is only partially translated (got bored). But the important stuff is once again there and correct.

 

Patent DD287053A5

PROCESS FOR OBTAINING BAEOCYSTIN BY BIOTRANSFORMATION OF TRYPTAMINE

 

 

Claims:

1. A process for the production of beocystin by biotransformation of tryptamine, characterized in that the aqueous solution of tryptamine with the addition of
methionine, phosphates and yeast extract under sterile conditions with basidiospores of higher fungi, such as those of the genera Psilocybe, Conocybe, Panaeolus and
Gymnopilus is inoculated and after the enzymatic biotransformation, the Baeocystin is separated in known manner by evaporation, extraction or chromatography.

2. The method according to claim 1, is characterized in that the biotransformation at 1 to 30*C depending on the type of fungus and with or without the addition of
antibiotics to the medium within 14 days.

Field of application of the invention

The invention relates to a process for the production of Baeocystin by biotransformation of precursors by basidiospores. The substance Baeocystin is a pharmaceutically
active substance and can be used as a model substance in basic neurochemical research.

Characterization of the known technical solution

The tryptamine derivative Baeocystin(4-phosphoryloxy-N-methyltryptamine) is found as an accompanying alkaloid of psilocybin and psilocin in naturally occuring
higher mushrooms of various genera in small to very small quantities.
It is well known that the alkaloid can be isolated from the extracts of natural fungi by using chromatographic methods (Pharmazie 1985, 274).
The method described has several disadvantages. The yield of alkaloid is only small because simple operating methods do not allow for separation of psilocybin
which has very similar physiochemical properties, chemically differing only by an additional methyl group. In addition the naturally occurring mushrooms are not
economically accessible. The biotransformation of tryptamine is known in cultured mycelia of higher fungi(DD-PS 265636) 0.25% calculated for the separation can then
take place only under complex separation methods with loss of yield. Other mycelial ingredients(e.g.lipids) require additional work up. Finally, the cultivation of
the mycelia takes at least 30 days in the biotransformation of tryptamine.

Object of the invention

The aim of the invention is to develop a simple and less expensive process for the production of baeocystin by biotransformation of tryptamine, which is in aqueous
solution with a short reaction time, achieves a high yield of Baeocystin without disturbing the accompanying alkaloids and organic substances.

Explanation of the nature of the invention

The inventions object is to select such media for biotransformation of tryptamine, which in aqueous solution ensures rapid formation of Baeocystin in high yield
without additional alkaloids.
According to the invention the object is achieved in that the aqueous solution of tryptamine with the addition of methionine, phosphates and yeast extract under
sterile conditions with Basidiospores and higher fungi, such as the genera Psilocybe, Baeocystin in a conventional manner by evaporation, extraction or
chromatography.
In the biotransformation, a selective hydroxylation and phosphorylation takes place in the 4-position on the indole ring of the tryptamine, at the same time the
amino nitrogen is mono-methylated on the side chain.
For the culture media solutions of 1.5 to 3% methionine, 1 to 4% tryptamine or its water-soluble salts, 1.5 to 3% soluble phosphate, 0.1 to 0.3% yeast extract and
water to 100%, also the addition of antibiotics if appropriate, basidiospores are transferred in an amount of 0.1 to 0.2%.
Fungal spores of the genera Psilocybe, Panaeolus, Gymnopilus and Conocybe are used for the biotransformation reaction. After a reaction time of 5 to 14 days at 1 to
30*C, the Baeocystin is isolated in a known manner by filtration, evaporation, preferably in vacuo and subsequent extraction and/or chromatography in a yield of 70 to
90%.
The invention will be explained in more detail by the following example.

Embodiment

Basidiospores of Psilcybe cubensis(Earle) Sing. were used for inoculation, which were obtained from the lamellae of mature fruiting bodies by release under sterile
conditions in a humidity chamber.
0.1g of spore dust was added under sterile conditions to a mixture of 1.5g methionine, 2g tryptamine hydrochloride, 1.5g KH2PO4 and 0.1g yeast extract in 95ml water
in a 300ml Erlemeyer flask with a cotton plug. After stirring for 10 days(magnetic stirrer) at 20*C, the bluish solution was filtered after 150ml of water had been
added. It was then extracted by shaking with 50ml of chloroform, the aqueous phase evaporated(vacuum), 20ml of methanol at 0*C added, filtered, the residue obtained
dissolved with 50ml of acetone and then with 50ml of acetone/diethyl ether and the solid residue dissolved with 500ml of methanol. After filtration to remove the
inorganic salt is the methanolic solution of Baeocystin i.Vak. eingedunstet.
Yield of Baeocystine after recrystallization in n-butanol: 2.1g.



#43 PistolPete13

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Posted 18 December 2018 - 11:32 PM

DD273449(A1) -1989- VERFAHREN ZUR GEWINNUNG VON 4-SUBSTITUIERTEN INDOLVERBINDUNGEN

 

METHOD FOR OBTAINING 4-SUBSTITUTED INDOLE COMPOUNDS

 

 

Example 1
A spore culture of the subtropical species Psilocybe cubensis (Earle) Sing. Was grown on 6% malt agar(agar: 2%). After 3 weeks a piece of mycelium about 2cm x 2cm was
aseptically taken and shaken in a 100ml sterile flask containing 20g of saddle filling in addition to 20ml of water for 10min, whereby the mycelium is crushed under the
filling materials generating a fine suspension.
Each 5 ml of the suspension is used aseptically to inoculate an autoclaved mixture of 50g of dry cow dung and 25g of rice grains in 80ml of water which still contains
0.5g tryptamine hydrochloride. After 14 days the culture medium was completely overgrown with white mycelium, after 20 days the fruiting bodies began to form. After 5
fructification waves were obtained from the defatted by petroether methanolic solutions by column chromatography on cellulose isolated with water-saturated n-butanol.
Yield: 21g Dried mushrooms Psilocin content:   3.3%
                                                 Psilocybin content: 0.02%

Example 2

In analogous procedure as indicated in Example 1 and using 60g of dry horse manure, 20g rice grains and 100ml of water with the addition of 0.25% Tryptamine
hydrochloride developed 29g of dried mushrooms containing psilocin 2.1%. In addition, the mushrooms contained 0.15% psilocybin.

 

Just a note saddle filling is actually saddle packing, the ceramic berl saddles seen in labs. Some people use a marble to break up the mycelium, exact same thing.



#44 coorsmikey

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Posted 19 December 2018 - 11:27 AM

I've failed to see the relevance of these abstracts to Using DMT to increase alkaloids in mushrooms. But none the less it is still interesting and thanks for translating it.



#45 PistolPete13

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Posted 19 December 2018 - 03:28 PM

Yeah sorry about that, I should have just put the smaller Baeocystin and Tryptamine abstracts up because it may be of some use in this thread. I also should have put a warning up, that it has been known in the past that when a researcher thinks he has discovered something that he rushes out and patents it in every way he can think of. Sometimes without peer review, adequate testing ect. so I would take what is read as 'leads to investigate further' more so than absolute fact. Oh yeah and sorry again for the thread de-rail! :blush:



#46 SteampunkScientist

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Posted 20 December 2018 - 07:55 PM

I'll have to try that rice trick with my Coffee Coir Tek (warfrats Tek). Might get some impressive yields!




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