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Offlineiudexk
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Registered: 08/18/01
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Loc: UK
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Increasing alkaloid levels
    #374192 - 08/18/01 06:26 PM (16 years, 7 days ago)

I saw the article in the cultivation section of this site where someone added d. illinoensis rootbark to increase alkaloid levels. However, it seems to me this is not an ideal method as

1) It is rootbark, and as such contains only a small amount of useful alkaloid (DMT) that can be metabolised(?) to psilocybin
2) D. illinoensis rootbark is quite expensive
3) It's a waste of DMT

However, if it works with DMT, surely it would work with straight L-tryptophan (cheap)?



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Offlineiudexk
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Re: Increasing alkaloid levels [Re: iudexk]
    #374201 - 08/18/01 06:47 PM (16 years, 7 days ago)

I did look at the alkaloid potentiation section of the MicoDirectory 2.0 post, but many of the links did not work and I could find no information on potentiation with L-tryp.



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OfflineTamrylin
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Re: Increasing alkaloid levels [Re: iudexk]
    #374204 - 08/18/01 06:55 PM (16 years, 7 days ago)

This all looks like boosting the alkaloid content with DMT, etc.. I read that hydrogen peroxide helps to form alkaloids because of its oxygen content.. Mycelium (all) is compatible with hydrogen peroxide, and it also happens to help prevent contamination.. Im testing to see if the shrooms get a bigger kick with added hydrogen peroxide... another week or two at most.. Why not try getting the mycelium to make more of its own psilocybin/psilocin? I got some of the info bout hydrogen peroxide at www.mycomasters.com .. ordered both volumes of the thin as hell guide.. some useful info in it



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OfflineAzure
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Re: Increasing alkaloid levels [Re: iudexk]
    #374323 - 08/19/01 12:57 AM (16 years, 7 days ago)

the purpose of having toxins(psilocybin, psilocyin, baeocystin, etc) in the first place is to deter predators from inhibiting the reproduction of the psychoactive mushroom. When the mushroom experiences less than optimal conditions during the fruiting cycle, it increases the amount of toxins in its tissue to protect its future generations. To mimic nature, drop the temperature 5-10 degrees below optimal fruitng temperatures to raise the drug content after pinning has occurred. Overall mass will be reduced, but overall drug content will be increased significantly.



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Offlinejonnyshaggs420
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Re: Increasing alkaloid levels [Re: iudexk]
    #374354 - 08/19/01 02:31 AM (16 years, 7 days ago)

1) The majority of the DMT is contained within the rootbark, with far less significant amounts in the rest of the plant.

2)Desmanthus root bark is not expensive if you do some comparative shopping, but Mimosa hostillas rootbark is a far better buy.

3)Yes its a waste of DMT.

Tests have been done on the absorbtion rate of DMT by psilocybe mushrooms. The articles are linked somewhere else in this forum. DMT is very, very poorly absorbed by mushrooms and so is L-tryptophan. On the other hand tryptamine is easily absorbed and converted into useful alkaloids. Possibly this is due to water solubility. And as a side note during the major discussion of this topic a few months ago, it was found that fully supplementing a mushrooms diet will increase alkaloid content of the crop far better and cheap than just adding tryptamines, and results showed that mushrooms with only added tryptamine grow small and very twisted.

Novelty


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Offlineiudexk
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Re: Increasing alkaloid levels [Re: jonnyshaggs420]
    #374453 - 08/19/01 08:24 AM (16 years, 7 days ago)

hmmmmmmm ok thankyou everyone. BTW in case anyone doesn't know, L-tryptophan can bee very easily decarboxylated to tryptamine...



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Offlineiudexk
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Re: Increasing alkaloid levels [Re: iudexk]
    #374454 - 08/19/01 08:34 AM (16 years, 7 days ago)

L-tryptophan $185/455g
D. illinoensis rootbark $75/455g
M. hostilis rootbark $189/455g

1lb of L-tryptophan would make about 400g of pure tryptamine, 1lb of D. illinoensis rootbark would contain about 5g at most DMT.



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Offlineiudexk
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Re: Increasing alkaloid levels [Re: iudexk]
    #374578 - 08/19/01 04:22 PM (16 years, 6 days ago)

entered: Jan 10 1997 by Psylocybe Fanaticus - Seattle
In the following transcription of the science paper and discovery of Dr. Jochen Gartz, he describes adding a 25 millimolar concentration of Tryptamine HCL (a psilocin and psilocybin precurser) to the Cubensis substrate and under lab control conditions, discovered the potentiation of psilocin into never before measured levels in Cubensis fruitbodies of up to 3.3% psilocin which is several times the potency as regular Cubensis.

PF TEK application of the Gartz Tryptamine tecknique.
1/2 pint jar:

1/2 - 2/3 cup of vermiculite + 1/8 cup of brown rice powder and (45 milliliters of water with .16 grams of Tryptamine HCL added)

PF experiment results:

The fungus cultured as usual except that the fruitbodies grew dwarfed. Bioassay showed that they are at least 3 times the usual potency.

--------------------------------------------------------------------------------

March 6 1997 entry - Tryptamine formula update
The above formula is a bit to much for the pf tek. That is why the shrooms grew dwarfed and some jars failed to fruit. The answer is that the pf substrate is much lighter and thinner than Gartz' substrate. Gartz used cooked brown rice and cow dung. This is heavier, thicker and more nutritious than the pf substrate formula, so therefore, the tryptamine hcl content should be less also. PF has received some very reliable information that 1/2 to 3/4 of the above formula should be used. So instead of .16 grams of tryptamine hcl, use .1 or less grams of tryptamine hcl.

--------------------------------------------------------------------------------

Planta Medica 55 (1989) page 249 - 250 Jochen Gartz
BIOTRANSFORMATION OF TRYPTAMINE IN FRUITING MYCELIA OF PSILOCYBE CUBENSIS.

Jochen Gartz
Institute of Biotechnology, Academy of Sciences of the GDR,
Permoserstrasse 15, GDR-7050 Leipzig, German Democratic Republic

Received: March 13, 1988

ABSTRACT

Mycelial cultures of Psilocybe Cubensis, with the ability to form psilocybin and psilocin de-novo, also hydroxylated and methylated fed tryptamine to give psilocin in up to 3.3% dry mass of the obtained fruit bodies. By using HPLC and TLC, it was found that these mushrooms contain only a small amount of psilocybin (0.01-0.2% dry mass). The values of psilocin are the highest described in any mushrooms.

INTRODUCTION

Psilocybe Cubensis (Earle) Sing, is a substropical mushroom and contains the indole alkaloid psilocybin and only small amounts of its dephosphorylated counterpart psilocin (1-4). Variations in these metabolites have been well demonstrated by investigations of fruit bodies cultivated under controlled conditions of a rye-grain medium (2) and rice substratum (3), respectively.

The study of psilocybin biosynthesis in submerged culture of P. Cubensis showed that radioactive tryptamine functioned as a better precurser than tryptophan (5-7). It was found that not less than 22.4% of the psilocybin formed was derived from the labelled presursor tryptamine (5). The level of psilocin was generally zero in the mycelial tissue from these experiments (5-7).

In the present paper, the biotransformation of fed tryptamine in fruiting mycelia of P. Cubensis is described.

MATERIALS and METHODS
Cultivation of Psilocybe Cubensis

A dried cow dung/rice-grain mixture (2:1) with twice the amount of water was used to obtain fast fructifications without casing of a strain (3) of P. Cubensis. A 25 mM concentration of tryptamine (as hydrochloride) was added to this medium. Cultivations without the addition of tryptamine were also tested. The methods of cultivations were described in (3).

The first sporocarps were produced by cultures of P. Cubensis in 3 to 4 weeks. The cultures continued to produce mushrooms in five flushes. Each flush was harvested as soon as the sporocarps were mature. The mushrooms were immediately freeze-dried, sealed in plastic, and stored at -10 degrees C until analysis.

EXTRACTION and ANALYSIS The extraction procedure and the analysis of the indole alkaloids by using HPLC and TLC were described in the previous papers (3,8-10). The presence or absence of tryptamine was demonstrated by TLC as described by Stijve et al. (11).

RESULTS and DISCUSSION

The cow dung-rice mixture actually produced the first flush of mushrooms earlier than the cultivations on ry (with casing) (2) and rice (3), respectively. They yielded an average of 3 g dry mass per 10 g substratum.

Under the same culture conditions, the fructification times, the yields, and sizes of the mushrooms as well as the blueing feature (3) were equal when the growth media also contained high concentrations of tryptamine. Initial experiments without the addition of tryptamine were performed to determine the content of psilocybin and psilocin in comparison with experiments using other culture conditions and/or media (2,3).

The levels of psilocybin and psilocin varied from one flush to the next, but generally were much the same as those in the other experiments (2,3) (table 1). Consistently low levels of psilocin were found in the mushrooms without the addition of tryptamine to the substratum. Additionally, psilocin generally was absent in the first flush as was also observed in earlier investigations (2,3). Table 1 shows that the fed tryptamine gives high values of psilocin in each flush from the cultures.

Table 1 Variation of psilocybin and psilocin levels in P. Cubensis as a function of flush number from the cultivations with (a) and without (b) addition of tryptamine (25 mM concentration).

Flush no. Psilocin Psilocin Psilocybin Psilocybin
a b a b
1. 2.1 - 0.01 0.55
2. 3.3 0.01 0.02 0.48
3. 2.8 0.02 0.2 0.51
4. 3.1 0.09 0.07 0.46
5. 2.9 0.15 0.13 0.61


These psilocin levels are uncommonly high (from 2.1 to 3.3%) since values reported for psilocin in dried mushrooms are always below 1% (1-4,12,13).

Inocybe Aeruginasens Babos contains only traces of psilocin but high amounts of the incompletely methylated psilocybin (baeocystin) (9). In contrast to the intitial experiments without an addition of tryptamine, the mushrooms generally contained only small amounts of psilocybin. The tryptamine level was always zero in each mushroom. In this case no tryptamine was additionally found in the methanolic extract of the vegetative mycelia from the substratum.

In a previous report, Gartz (3) was unable to detect baeocystin in P. Cubensis. But Repke et al. (14) reported traces of baeocystin in other strains of P. Cubensis about 10 years ago. They suggested that many non-specific enzyme systems exist in fungi which have the ability to oxidise exogenously added compounds, as well as normal, obligatory intermediates (14).

The results in Table 1 show that the enzyme systems in P. Cubensis have a high hydroxylation and methalation capacity to convert added Tryptamine to psilocin. It is possible that a reduced amount of phosphate in the culture media decreased the biosynthesis of psilocybin from psilocin in the media.

P.Cubensis also failed to produce detectable amounts of baeocystin under these culture conditions.

Acknowledgements

The author thanks the following persons: G. Drewitz, T. Stijve, G.K.Muller, and M. Gey who generously supplied valuable information.

REFERENCES


Heim, R., Hoffman, A. (1958) Compt. Rend. 247,557.
Bigwood, J.. Beug, M.W. (1982) J. Ethnopharm. 5, 287.
Gartz, J. (1987) Beitrage zur Kenntnis der Pilze Mitteleuropas 3, 275.
Badham, E. (1984) J. Ethnopharm. 10, 249
Agurell, S., Blomkvist, S., Catalfomo, P. (1966) Acta Pharm. Suecica 3, 37.
Agurell, S., Nilsson, J.L.G. (1968) Acta Chem. Scand. 22, 1210.
Agurell, S., Nilsson, J.L.G. (1968) Tetrahedron Lett. 1063.
Gartz, J. (1985) Pharmazie 40, 134.
Gartz, J. (1987) Planta Med. 53, 539.
Semerdzieva, M., Wurst, M., Koza, T., Gartz, J. (1986) Planta Med. 52, 83.
Stijve, T., Hischenhuber, C., Ashley, D. (1984) Z. Mykol. 50, 361.
Beug, M.W., Bigwood, J. (1982) J. Ethnopharm. 5, 271.
Ohenoja, E., Jokiranata, J., Makinen, T., Kaikkonen, A., Airaksinen, M.M. (1987) J. Nat. Prod. 50, 741
Repke, D.B., Leslie, D.T., Guzman, G. (1977) Lloydia 40, 566.

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Offlinejonnyshaggs420
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Re: Increasing alkaloid levels [Re: iudexk]
    #374930 - 08/20/01 02:22 AM (16 years, 6 days ago)

yep those be some of the articles.

How would one go about decarboxylating this L-tryptophan?

Novelty


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Offlineiudexk
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Re: Increasing alkaloid levels [Re: jonnyshaggs420]
    #375024 - 08/20/01 07:02 AM (16 years, 6 days ago)

80g L- or D,L-tryptophan is dissolved in 250mL turpentine (or better cyclohexanol), and 5mL thuja plicata essential oil, or better an equivalent amount of a high-boiling ketone (e.g. cyclohexanone) is added. The solution is then heated to 150?C. Bubbles (CO2) will appear. When no more bubbles evolve, stop heating. This will take between 1.5 and 4hrs, possibly more. The solution is then allowed to cool, washed with a 5% aqueous solution of baking soda, and then twice with distilled water. It is then extracted with 3 portions of 10% HCl (Spirits of Salt drain cleaner is 32% HCl). The extracts are combined, washed with lighter fluid, then basified to pH 7 with a 20% aqueous NaOH soln. (Caustic Soda drain cleaner is 99% NaOH), then washed again with more lighter fluid. The solution is then further basified to pH 14 with 20% aq. NaOH and extracted with portions of lighter fluid which are combined, dried by filtering through powdered anhydrous magnesium sulphate (Epsom salts dried in oven @250?C for 4hrs, then crushed up). Anhydrous HCl gas (generate from salt + conc. sulfuric acid drain cleaner in a plastic bottle with a tube in the top) is then passed through the solution while stirring until the pH is 6.5 when checked with pH paper. The solution is sealed and left in a freezer overnight. Filter off the trpytamine HCl crystals in the morning, and wash them with a little more lighter fluid. Estd. yield: ~85%

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Offlineonetone
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Re: Increasing alkaloid levels [Re: iudexk]
    #375031 - 08/20/01 07:15 AM (16 years, 6 days ago)

MAKING TRYPTAMINE

An article from a 1996 copy of the Entheogen Review:, copied from Gnosticgarden.com

QUOTE:
--------------------------------------------------------------------------------

Making Tryptamine From Milk

Here's an idea on boosting the Psilocin/Psilocybin content of P. cubensis. I read in some of J. Gartz's work, that he could increase the potency of P. cubensis up to six fold by feeding the fungus Tryptamine Hcl. This substance is not easily obtained, but tryptamine can be created via bacterial action on tryptophan-containing substances. Milk is a common source of tryptophan (some people think that this is why a warm glass of milk makes people sleepy). The best conversion claim of tryptophan to tryptamine via bacterial action was from 75-85%. An average glass of milk contains 35-65mg of tryptophan, a gallon from about 600-1000mg. This could be a ready-made source of from 400-800mg of tryptamine, just from sour milk. A local grocer will sell me past-dated milk for 75 cents a gallon. I would suspect that many grocery stores might even give it away. The best way to add the putrified milk solids to the mushroom culture would be to:

1) Let it totally spoil (do this where the smelll doesn't matter), and then dry it to a solid.

2) Sterilize the solid in a pressure cooker for the same time and pressure settings as for agar.

3) When innoculated grain is ready to put into trays, carefully mix in a S.A.C supplemental feeding at casing: a procedure described by Paul Stamets in his bug blue mushroom bible. Add an amount equal to ten percent of the grain/substrate weight and mix both thoroughly. This would be cheap, and most important, invisible to the police state - no chemical house requisition paper trails left.




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Offlineiudexk
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Re: Increasing alkaloid levels [Re: onetone]
    #375038 - 08/20/01 07:28 AM (16 years, 6 days ago)

My method is also OTC and gives pure tryptamine HCl that can bee accurately measured =)

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OfflineHumidity
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Re: Increasing alkaloid levels [Re: Azure]
    #375142 - 08/20/01 01:08 PM (16 years, 6 days ago)

Azure

That is a good theory about the toxins being used to deter predators, this may or may not be true.

The information about the temperature playing a important role in increasing psilocin/psilocybin production may be true. I have done some reading about this kind of stuff and the enviromental conditions do affect the amount of metabolites that are formed.

Have you confermed stronger mushrooms with lower temps. or did you just read this somewhere???



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Offlinegray1
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Registered: 04/30/01
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Re: Increasing alkaloid levels [Re: iudexk]
    #375639 - 08/21/01 02:33 PM (16 years, 5 days ago)

Azure,
what makes you think that inhibiting animals from eating the mushrooms is the purpose of psilocybin/psilocin anymore than enticing animals to eat them and in doing so encouraging widerspread propogation? it is plausible that psilocybin is toxic to insects, bacteria, other parasitic fungi, viruses, etc... but it certainly isn't toxic to mammals.

Iudexk,
using a DMT containing substrate like bark or grass is most likely wasteful because the DMT is not biologically availiable to the organism.

if you search this forum for increased potency, tryptophan, dmt, m. hostilis rootbark in mailbox, etc... you will find that these issues have been discussed at length earlier in the year and the general consensus to this point has been that it may be possible for substrate additions to increase potency in theory, it is unlikely that there will be any actual increase in potency, and even if there is, there is no accurate way to determine an increase in potency.
addition of pure tryptamine may be an exception to the rule, and worth further experimentation with both techniques described above (organic synth and biosynth, though not everyone has the equipment/supplies to do the orgo conversion, also, in the US, tryptophan is not legal and therefore must be purchased from abroad and imported or acquired through a source that is Just Less Fucking legitimate...

increasing potency is certainly a worthy pursuit for those inclined, but it is most likely more efficeint to produce more quantity of fruit bodies.


how do you write bold text, colors, etc...
there really ought to be a tutorial for bulletin board novices like me.


gray1

"i am a citizen of the world" diogenes


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Offlineun0r
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Re: Increasing alkaloid levels [Re: gray1]
    #375740 - 08/21/01 06:26 PM (16 years, 4 days ago)

Anyone have an idea if tryptamine (freebase) would be just as effective as tryptamine hcl? Anyone have experience with this?

- Bill Goat


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InvisibleMr. Pink
Model Prisoner
Registered: 01/30/01
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Loc: Small town with a BIG pro...
Re: Increasing alkaloid levels [Re: un0r]
    #375949 - 08/22/01 12:13 AM (16 years, 4 days ago)

In reply to:

Just Less Fucking legitimate




LMAO! Good one..



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Offlinejonnyshaggs420
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Re: Increasing alkaloid levels [Re: gray1]
    #375974 - 08/22/01 12:56 AM (16 years, 4 days ago)

Tryptophan is illegal? Wow, GNC is going down.

Novelty


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Offlineiudexk
member
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Re: Increasing alkaloid levels [Re: un0r]
    #376145 - 08/22/01 05:44 AM (16 years, 4 days ago)

As tryptamine freebase is not water-soluble, I think it would be much less effectively absorbed (if at all).

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Offlinetom
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Re: Increasing alkaloid levels [Re: iudexk]
    #376219 - 08/22/01 09:21 AM (16 years, 4 days ago)

So if one were to add psilocin to the substrate would that increase psilocin levels in the fruit bodies? Silly question but it has serious answers. Not something for most of us to try... just the hard core scientists.

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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
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Re: Increasing alkaloid levels [Re: gray1]
    #377523 - 08/24/01 03:34 AM (16 years, 2 days ago)

In reply to:

what makes you think that inhibiting animals from eating the mushrooms is the purpose of psilocybin/psilocin anymore than enticing animals to eat them and in doing so encouraging widerspread propogation? it is plausible that psilocybin is toxic to insects, bacteria, other parasitic fungi, viruses, etc... but it certainly isn't toxic to mammals.



In animal trials where said animals were able to self-administer various drugs, eg nicotine, cocaine, morphine, said animals would continue to self-administer the drug to the point of unconsciousness/death. Whereas similar trials involving hallucinogens resulted in little to no self-administration.
Animals (except some humans;) don't seem to enjoy hallucinogens.

Old Donkeys don't.......oh. Nevermind.
Drool Donkey Island
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