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Offlineiudexk
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Registered: 08/18/01
Posts: 107
Loc: UK
Last seen: 22 years, 2 months
Increasing alkaloid levels
    #374192 - 08/18/01 04:26 PM (22 years, 6 months 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 04:47 PM (22 years, 6 months 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 04:55 PM (22 years, 6 months 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 - 08/18/01 10:57 PM (22 years, 6 months 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 12:31 AM (22 years, 6 months 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 06:24 AM (22 years, 6 months 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 06:34 AM (22 years, 6 months 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 02:22 PM (22 years, 6 months 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.

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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.

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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 12:22 AM (22 years, 6 months 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 05:02 AM (22 years, 6 months 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 05:15 AM (22 years, 6 months 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 05:28 AM (22 years, 6 months ago)

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

http://www.magickherbs.co.uk/


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OfflineHumidity
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Re: Increasing alkaloid levels [Re: Azure]
    #375142 - 08/20/01 11:08 AM (22 years, 6 months 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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Re: Increasing alkaloid levels [Re: iudexk]
    #375639 - 08/21/01 12:33 PM (22 years, 6 months 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 04:26 PM (22 years, 6 months 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: 01/29/01
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Re: Increasing alkaloid levels [Re: un0r]
    #375949 - 08/21/01 10:13 PM (22 years, 6 months 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 - 08/21/01 10:56 PM (22 years, 6 months ago)

Tryptophan is illegal? Wow, GNC is going down.

Novelty


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Offlineiudexk
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Re: Increasing alkaloid levels [Re: un0r]
    #376145 - 08/22/01 03:44 AM (22 years, 6 months 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 07:21 AM (22 years, 6 months 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.

"I got that Fiya Powda" -- some black dude

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InvisibleChampion des Champignons
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Re: Increasing alkaloid levels [Re: gray1]
    #377523 - 08/24/01 01:34 AM (22 years, 6 months 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
Support the FSR!


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Offlinealuminum_can
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Re: Increasing alkaloid levels [Re: iudexk]
    #377763 - 08/24/01 12:11 PM (22 years, 6 months ago)

i have found a nice way to increase the alkaliod levels. it is really a quite simple procedure and here it is. just eat more!!!! shrooms are pretty fucking abundant if you are growing them yourself which im guessing that you are so just eat more, if you want to trip harder. i dont see how soaking verm in dmt will increase the potency of youre shrooms, just waste dmt and time and money. and as im told time and money are two of the most important things to worry about these days, so you should be careful what you do with them. increasing the alkaliod level would be like telling you to grow grow a different hair color or something. genetics are already decided for the shroom and it would be hard, or even impossible to change it. i dont know much about the science of shrooms metabolism and the way they these fruitbodies just pop out of a "ball" of fungus either. it blows my mind.

one plus one plus one equals three


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Offlineiudexk
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Re: Increasing alkaloid levels [Re: aluminum_can]
    #378456 - 08/25/01 10:48 AM (22 years, 6 months ago)

increasing the alkaliod level would be like telling you to grow grow a different hair color or something. genetics are already decided for the shroom and it would be hard, or even impossible to change it.

It is well known that alkaloid levels in various plants can be manipulated. mescaline etc.

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Offlinegray1
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Re: Increasing alkaloid levels [Re: iudexk]
    #379772 - 08/27/01 07:08 AM (22 years, 6 months ago)

true dat.


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OfflineelfstoneS
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Re: Increasing alkaloid levels [Re: iudexk] * 2
    #26312685 - 11/10/19 09:42 PM (4 years, 3 months ago)

Adding oat bran as a supplement to the substrate, which is high in phosphorous, may serve to increase the psilocybin production (4-phosphoryloxy-dimethy,tryptamine) which is stable at room temperature and assures more lasting potency when the mushrooms are dried.  I know of no studies that confirm this hypothesis but anecdotally it seems to result in very potent Ps. mexicana mushrooms, which, per Stamets psilometric scale, is high in psilocybin and low in psilocin and has no baeocystin.  The Ps. mexicana also do not bruise blue, which is seen in mushrooms, such as Ps. cubensis, which has a higher psilocin content.


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OfflineSchroomfairy
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Re: Increasing alkaloid levels [Re: elfstone]
    #26342590 - 11/24/19 02:14 PM (4 years, 3 months ago)

Ah, I love the bringing back of old posts. So, 18 years later has anything changed? Eflstone, you mentioned oat bran... I see you mentioned no studies but has this been tested by another grower?

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InvisibleStygianKnight
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Re: Increasing alkaloid levels [Re: Schroomfairy] * 1
    #26345336 - 11/25/19 07:06 PM (4 years, 3 months ago)

A lot has changed, and we now know that there’s nothing you can add to your substrate that will increase or decrease potency.

Cubes use ATP or Adenosine TriPhosphate as energy storage, while they may indeed like more phosphate, an increase in phosphate isn’t going to increase psilocybin production as phosphate isn’t the limiting factor.  Nor is tryptophan a limiting factor either.

We also know that Norbaeocystin and Baeocystin are intermediary steps in the organic synthesis of psilocybin and thus all species that use this pathway contain trace amounts.  Psilocin is a bit different as it it’s not in the synthesis but from exposure of psilocybin to PsiK a dephosphorelating enzyme.

Well so if none of that works, what does? 
Genetics.
The main limiting factor is the efficiency, speed and quantity of the enzymes that build psilocybin and its genetics that codes for the proteins that build the enzymes that builds the magic.  It’s that set of enzymes that takes common building blocks and puts them together in the right ways and those enzymes that E.coli formed when the psilocybin producing genetics was shot into it.

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Re: Increasing alkaloid levels [Re: StygianKnight]
    #26346885 - 11/26/19 02:16 PM (4 years, 3 months ago)

Quote:

StygianKnight said:
A lot has changed, and we now know that there’s nothing you can add to your substrate that will increase or decrease potency.

Cubes use ATP or Adenosine TriPhosphate as energy storage, while they may indeed like more phosphate, an increase in phosphate isn’t going to increase psilocybin production as phosphate isn’t the limiting factor.  Nor is tryptophan a limiting factor either.

We also know that Norbaeocystin and Baeocystin are intermediary steps in the organic synthesis of psilocybin and thus all species that use this pathway contain trace amounts.  Psilocin is a bit different as it it’s not in the synthesis but from exposure of psilocybin to PsiK a dephosphorelating enzyme.

Well so if none of that works, what does? 
Genetics.
The main limiting factor is the efficiency, speed and quantity of the enzymes that build psilocybin and its genetics that codes for the proteins that build the enzymes that builds the magic.  It’s that set of enzymes that takes common building blocks and puts them together in the right ways and those enzymes that E.coli formed when the psilocybin producing genetics was shot into it.





Thank you so much for your thoughtful reply. That clarified a lot of things for me. So the next question is your final statement in regards to the genetics. Even if we take a print and put it on agar with as many transfers as needed to obtain the perfect plate, is that the way to the perfect genetics?

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OfflineMycoactive
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Re: Increasing alkaloid levels [Re: Schroomfairy]
    #26346913 - 11/26/19 02:41 PM (4 years, 3 months ago)

There are a few things to consider when it comes to increasing alkaloid levels:

1. You can increase overall alkaloid levels by increasing yield. Yield is dependent on many factors (genetics, substrate composition, temperature, moisture levels, etc.).

2. You can increase alkaloid levels per unit of biomass by genetic selection. You can also grow species that reliably produce high alkaloid content.

3. As StygianKnight suggested, psilocybin-producing fungi are chemically complex. There are molecules other than psilocybin/psilocin that likely contribute to the physical and psychological effects that you experience when you consume mushrooms. Increasing psilocybin/psilocin content may come at the expense of other tryptamines that have a positive effect on the psychedelic experience.

My point is that there are many different ways to potentially increase alkaloid levels, but it's important to understand that psilocybin/psilocin are not the only alkaloids present in psilocybin-producing fungi.

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Re: Increasing alkaloid levels [Re: Mycoactive] * 1
    #26349473 - 11/27/19 08:22 PM (4 years, 3 months ago)

As Mycoactive hit on, what are we talking about when we say increasing levels? Is that overall amount of alkaloids or ratio of alkaloids to fruit mass?

Getting a clean sample will indeed help to produce a larger amount of alkaloids than a dirty sample, but that’s not quite the same here since the solution to weak genetics, is just a larger dose.

Stronger genetics as in a higher amount of alkaloid to fruit mass comes about in a number of different ways, many of which we can’t measure easily at home.  So for at home hobbyist work the suggestion is to find the best growing fruits and clone them.  Take whatever is happiest in your environment and give it more room to grow.

When it comes to straight producing more enzymes it’s not known what the triggers are, or what causes production to slow down or stop. The issue is that enzymes are finicky and require specific local conditions to work their best.  Beyond just producing more enzymes it’s also about how much care is being taken by the mycelium to give them what they need to work efficiently. This is in part the reason for the suggestion above, it’s a presumption that fruits that are performing well in your environment are in their personal best environment to keep things stable inside themselves, and the enzymes working well.

Breeding genes that produce strong fruits with other genes that produce strong fruits can potentially help boost the production content, but it’s just dice rolls.  Just look at how famous Penis Envy and related strains are in part due to their potency increase.

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Re: Increasing alkaloid levels [Re: StygianKnight]
    #26360326 - 12/04/19 02:06 AM (4 years, 2 months ago)

I’ve wanted to try to reproduce Gartz’s experiment for years. Shulgin certainly thought there was something to it. Sadly at this point it reads mostly as a “wive’s tail” in terms of implementation. Further study needed.


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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26363611 - 12/05/19 06:09 PM (4 years, 2 months ago)

What experiment is that?

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Re: Increasing alkaloid levels [Re: StygianKnight]
    #26363765 - 12/05/19 07:37 PM (4 years, 2 months ago)

Quote:

StygianKnight said:
Nor is tryptophan a limiting factor either.





Correct me, but if no presents of tryptophan(at all) nothing to convert to Psilocybin?

Learn more about Psilocybin



I believe this this is why no one could reproduce Gartz's study. I think his dung had super low to almost no tryptophan. Where when others tried almost all the spawn/subs have some amount of tryptophan.

Yes adding tryptophan will NOT increase potency. This debates pisses off a lot of old hands. The way I look at it is, you can't force a cube to bio tryptophan that's all on genetics, but I want to make sure some is there to work with.

Please jump in to correct me if needed.

Edited by murderlabz (12/05/19 07:39 PM)

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Re: Increasing alkaloid levels [Re: Schroomfairy] * 1
    #26363830 - 12/05/19 08:20 PM (4 years, 2 months ago)

Quote:

Schroomfairy said:
What experiment is that?




It’s already been posted earlier in this thread. Gartz supposedly added tryptamine HCL to his substrate, and the result was each flush containing progressively, obscenely higher levels of psilocin in the fruit bodies (up to 3.3% in the third flush) compared to the normally grown mushrooms used as the “control.” Psilocybin levels were lower than in the control batch. This was in the late 80’s. To my knowledge it has never been duplicated. Here’s the original study:

https://pdfs.semanticscholar.org/e648/5b3f89815404f8e0912507aa693a68f27f80.pdf

Also related: Alexander Shulgin wrote in TiKHAL (late ‘90’s) how if one added DMT to the substrate the mushroom would simply take it and convert it into 4-HO-DMT (psilocin), yielding higher potency levels of psilocin. He also spoke of “very interesting research done in Germany” showing that if specific tryptamine are added to substrate, such as DIPT, DET, MET, etc, the mushroom will add it’s four hydroxylated (“4-HO”) signature to it and create psychedelic substances unknown in nature such as 4-HO-DIPT, 4-HO-DET, 4-HO-MET, etc, in addition to or in lieu of psilocin/psilocybin. Hamilton Morris recently spoke of this, citing pretty much word for word from Shulgin, on the Joe Rogan Experience.

I’m the fence on this one. I’ve not seen any studies verifying what Shulgin wrote, but haven’t researched it much either. My understanding is that it is at least “in part” correct. As a chemist, I can assure you that weird shit can happen in a laboratory setting for all sorts of reasons. A one off study doesn’t always mean a lot. So which part or how much is true...who knows. It’s worth doing some deep internet digging about. It’s likely there are at least some legitimate studies that have at this point been conducted, but I couldn’t say for sure. If it’s true it says a lot about what mushrooms are capable of as an active “pharmacological vessel.”

As an experiment, I once tried adding a tryptamine (melatonin) to my grains before PCing. It had no effect whatsoever to the final potency. Possible reasons are that 1) the tryptamine was “killed” off and rendered inactive during the process of sterilization 2) melatonin is incompatible here due to the fact that it is a tryptamine already chemically structured as a five hydroxylated (“5-HO”) substance, whereas tryptamine like the ones listed above are not 3) the notion of tryptamine supplementation increasing alkaloid levels is all just hocus pocus bullshit. Couldn’t say which.

It’s an interesting, weird, and largely unexplored subject. The whole “DMT in the substrate to increase alkaloid levels” idea was almost certainly something Shulgin started by writing about it in TiKHAL. It’s important to remember he was a chemist and not a grower, and it might all be more theoretical than he lets on. If it’s true, a specific set of circumstances and processes as to how exactly to supplement with DMT are probably needed - as far as to when, how much, etc. Like much of PiKHAL and TIKHAL, it’s designed to encourage experimentation.

Unlike some on this forum, I am personally absolutely certain that there are things that can influence potency aside from genetics, namely relating to nitrogen and carbon presence in certain ratios. There is actually a study from way back in 1968 that demonstrably shows this, if it is to be believed.  Whether for positive or negative, alkaloid levels of psilocin, psilocybin, baecystin, and norbaecystin were all affected. I’ll find it and post it.


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Edited by Psilotyl (12/05/19 09:33 PM)

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OfflineSchroomfairy
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Re: Increasing alkaloid levels [Re: Psilotyl] * 1
    #26363851 - 12/05/19 08:33 PM (4 years, 2 months ago)

Thank you for your excellent post. I will keep it for future reference.

When watching Hamilton Morris on the magic mushroom documentary, I found it interesting how the Mexican farmer talked about sugar cane and how it increased it's potency. Go to 14:48 [url=
Could be something to think about trying? He could be on to something? Or maybe even the possibility that adding to the substrate may actually cause something different. I am willing to complete some experiments for sure.

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Re: Increasing alkaloid levels [Re: Schroomfairy] * 1
    #26363901 - 12/05/19 09:07 PM (4 years, 2 months ago)

Tryptophan
Yes, if there wasn’t any tryptophan then psilocybin couldn’t be produced, but it’s just animals that can’t produce our own tryptophan, plants and microbes can.  Grain contains tryptophan and many fungi can that build tryptophan from smaller blocks.

Gartz (1989)
Can be presumed to be incorrect based on modern knowledge,
“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)” -Gartz

That the study got mostly psilocin and no psilocybin means something went wrong in their testing or prep.  The specific pathway has been found and it starts with phosphorylation, psilocin is only produced in mushrooms as an enzymatic breakdown of psilocybin into psilocin.  Besides it seems unwanted to get mushrooms with only the easily oxidized unstable compound.


DMT / DEPT / MET
Hypothetically yes adding DMT could reduce the amount of work the mycelium has to go through adding the two methyl groups, however it would need to be a sizable amount of DMT to end up in the right places without being oxidized by the oligermizing enzymes and you’re only skipping a single repeated step.

Similar with DEPT, etc. If you added enough that it found its way to the enzymes, they would probably do their work on them just fine, both the good ones and the bad ones.
This is something that we will probably see in the future but instead in psilocybin gene transferred e.coli bioreactors where the exact enzymes being produced can be controlled better.

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Re: Increasing alkaloid levels [Re: Schroomfairy]
    #26363926 - 12/05/19 09:29 PM (4 years, 2 months ago)

Quote:

Schroomfairy said:
Thank you for your excellent post. I will keep it for future reference.

When watching Hamilton Morris on the magic mushroom documentary, I found it interesting how the Mexican farmer talked about sugar cane and how it increased it's potency. Go to 14:48 [url=
Could be something to think about trying? He could be on to something? Or maybe even the possibility that adding to the substrate may actually cause something different. I am willing to complete some experiments for sure.




No problem. I think the sugarcane thing is entirely possible, “possible” being the key word there. Via YouTube, one can only guess what the nutritional contents of the soil actually are. If something is happening, the study I’m posting below would argue that it has to do with nitrogen and/or glucose levels. It’s the one I mentioned before, in my last post. Straight from MAPS:

https://bibliography.maps.org/resources/download/14256

I don’t want to discourage any experimentation, but I wouldn’t experiment with sugar alone in large amounts, if that’s what you’re saying. My current experiments are currently revolving around glycine/nitrogen supplementation. It’s too soon for me to say much but I have had one very encouraging result so far.


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OfflineNichrome
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Re: Increasing alkaloid levels [Re: StygianKnight] * 1
    #26363927 - 12/05/19 09:30 PM (4 years, 2 months ago)

Some interesting information on the evolution of psilocin and it's role in nature. https://www.biorxiv.org/content/biorxiv/early/2018/07/27/374199.full.pdf


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Re: Increasing alkaloid levels [Re: Nichrome] * 1
    #26364022 - 12/05/19 10:53 PM (4 years, 2 months ago)

Damn this is a good thread. Sorry if already posted but good stuff.

Full Paper -> Enzymatic Synthesis of Psilocybin

Abstract:
Psilocybinis the psychotropictryptamine-derivednatural product of Psilocybe carpophores,the so-called“magicmushrooms”.Although its structure has been known for 60 years,the enzymatic basis of its biosynthesishas remain edobscure. We characterized four psilocybin biosynthesisenzymes,namelyi) PsiD,which represents  class of fungall-tryptophan decarboxylases,ii) PsiK,which catalyzes the phospho transfer step,iii)the methyltransferase PsiM, catalyzing it erativeN-methyltransferas the terminal biosynthetic step,and iv) PsiH, amonooxygenase. Inacombined PsiD/PsiK/PsiM reaction, psilocybin was synthesized enzy-matically in a stepeconomic route from 4-hydroxy-l-trypto-phan. Given the renewed pharmaceutical interest in psilocybin, our results may lay the foundation for its biotechnological production

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


Full Paper -> Monoamine Biosynthesis via a Noncanonical Calcium-Activatable Aromatic Amino Acid Decarboxylase in Psilocybin Mushroom



Aromatic l-amino acid decarboxylases (AAADs) are a phylogenetically diverse group of enzymes responsible for the decarboxylation of aromatic amino acid substrates into their corresponding aromatic arylalkylamines. AAADs have been extensively studied in mammals and plants as they catalyze the first step in the production of neurotransmitters and bioactive phytochemicals, respectively. Unlike mammals and plants, the hallucinogenic psilocybin mushroom Psilocybe cubensis reportedly employs an unrelated phosphatidylserine-decarboxylase-like enzyme to catalyze l-tryptophan decarboxylation, the first step in psilocybin biosynthesis. To explore the origin of this chemistry in psilocybin mushroom, we generated the first de novo transcriptomes of P. cubensis and investigated several putative l-tryptophan-decarboxylase-like enzymes. We report the biochemical characterization of a noncanonical AAAD from P. cubensis (PcncAAAD) that exhibits substrate permissiveness toward l-phenylalanine, l-tyrosine, and l-tryptophan, as well as chloro-tryptophan derivatives. The crystal structure of PcncAAAD revealed the presence of a unique C-terminal appendage domain featuring a novel double-β-barrel fold. This domain is required for PcncAAAD activity and regulates catalytic rate and thermal stability through calcium binding. PcncAAAD likely plays a role in psilocybin production in P. cubensis and offers a new tool for metabolic engineering of aromatic-amino-acid-derived natural products.

--------------------------------------------------------------------
Full Paper -> Enzymatic route toward6-methylated baeocystin and psilocybin (2019)

Abstract
Methylated magic. The biocatalytic in vitro route to psilocybin was interrogated for substrate flexibility. 6‐Methylated substrates were accepted by the P. cubensis enzymes PsiD, PsiK, and PsiM and produced 6‐methylbaeocystin and 6‐methylpsilocybin. An in silico model of the methyltransferase PsiM identified nonconserved structural elements that likely govern the specificity of the methylation step.



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

Full Paper -> In vivo production of psilocybin in E. coli (2019)

Abstract
Psilocybin, the prodrug of the psychoactive molecule psilocin, has demonstrated promising results in clinical trials for the treatment of addiction, depression, and post-traumatic stress disorder. The development of a psilocybin production platform in a highly engineerable microbe could lead to rapid advances towards the bioproduction of psilocybin for use in ongoing clinical trials. Here, we present the development of a modular biosynthetic production platform in the model microbe, Escherichia coli. Efforts to optimize and improve pathway performance using multiple genetic optimization techniques were evaluated, resulting in a 32-fold improvement in psilocybin titer. Further enhancements to this genetically superior strain were achieved through fermentation optimization, ultimately resulting in a fed-batch fermentation study, with a production titer of 1.16 g/L of psilocybin. This is the highest psilocybin titer achieved to date from a recombinant organism and a significant step towards demonstrating the feasibility of industrial production of biologically-derived psilocybin.

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

Full Paper -> Biosynthesis of bioactive natural products from Basidiomycota (2019)

Abstract
The Basidiomycota, also called club fungi, comprise a diverse group of fungi. Basidiomycota are strongly related to ecosystem functioning along with human life. These fungi display a wide range of bioactivities, and some are known to produce of deadly toxins or hallucinogens. Some Basidiomycota have be used as medicinal mushrooms for thousands of years. Recently, the biosyntheses of several classes of natural products from Basidiomycota have been reported. Here, we review recent studies on the biosynthetic pathways and enzymes of bioactive natural products from Basidiomycota fungi, with a focus on terpenoids, alkaloids, ribosomally synthesized and post-translationally modified peptides (RiPPs), and polyketides.



Edited by murderlabz (12/05/19 11:22 PM)

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OfflineSolipsis
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Re: Increasing alkaloid levels [Re: murderlabz] * 1
    #26372448 - 12/10/19 06:32 AM (4 years, 2 months ago)

I haven't fully read murderlabz' wonderful contribution but from what I know it seems that it can be relatively pointless to try and change a part of metabolism by upping one precursor or metabolite concentration.
Because metabolism consists of many equilibria of catalyzed reactions and feeding this system doesn't guarantee at all that it will end up in a useful way.

So yeah it makes sense to work on the enzymatic activity. If psilocybin is a form of storage for an oligomerization reaction and metabolically speaking psilocybin is kinda cornered by mainly the phosphorylation/dephosphorylation.. possibly it would help to modify the rate of phosphorylation as this may build up psilocybin concentration without it being broken down again through another path (hopefully there isn't one im too lazy to check right now).

If quantative analysis was much easier I guess we could just breed and select for it as we may pick up on mutants being generated and we could stabilize them without them just being noticed as a batch of above average potency. Or who knows, someone might crispr it.. >_>

And maybe there are agents which can induce PsiK and change the equilibrium of catalysis? Idk if that would be easier than increasing the expression. What regulates dephophorylation?

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Re: Increasing alkaloid levels [Re: Solipsis] * 1
    #26372883 - 12/10/19 10:45 AM (4 years, 2 months ago)

Without sounding like a broken record, it’s important to remember that best way to achieve higher potency, or whatever you want with mushrooms, is indeed honing genetics. For instance, I have found the PES Hawaiian strain to be noticeably on the potent side, and worth further work. Cloning, agar isolation - these methods are tried and true. As RR said in Let’s Grow Mushrooms, agar work is great because it allows us to be “mad scientists.”

With that said, I am a believer that grain, substrate, fruiting conditions, and harvesting protocol all affect end potency as well. Just because it’s not been empirically proven to be so doesn’t mean we shouldn’t experiment, anyway.

Here is something interesting. Paul Staments recently found that the Polyphore mushroom extracts currently in development as a means of eradication of the “Varroa destructor” parasite and it’s RNA virus, the deformed wing virus (DWV) - which is the cause of widespread colony collapse among bees - have an efficacy of 10:1 when grown on corn, but 1000:1 ratio when grown on wood (I believe birch). The cause is of this is not known, nor has the active enzyme or whatever it is that is working so well to kill off these parasites and the RNA virus via the Polyphore mushroom even been identified...but it is known that SOMETHING in the substrate plays a part in greatly “up-regulating” it’s content. You can listen to him speak out this toward the end of his recent interview on the Joe Rogan Experience.

Now admittedly we are dealing with very different mushrooms when we start talking about psilocybes. But it still begs the question: why should they, at least to an extent, be any different? Is it not possible to manipulate them similarly? This is the core of scientific inquiry: questioning, asking the proverbial “what if” even when we barely know where to even start. & I think the question is worth pursuing.

I don’t argue with people who are adamant that substrate or supplementation techniques cannot effect potency. This has been their experience. And many like RR have damn good reasons as to why they hold to this. My differing opinion, for instance, that cubensis grown on manure are to some degree more potent than those grown on coir, is just that - an opinion. I have no empirical evidence. Which sucks, because locking in any empirical evidence here is damn near impossible.

One mush remember, no matter what your stance, that we are dealing with an unfairly illegal substance, one that has been under tight prohibition by power hungry bastards  across the globe for half a century (with the exception of Brazil and some others). This directly effects the ability to carry out proper research in a serious way even for scientists, and especially for the average, inquiring Joe. To amass any real data scientifically here just has not really been very possible. Thankfully all this is changing, slowly but surely. I have hope there will eventually be some real answers.

Murdelabz post gives us a lot to look at. I understand some of it but most of it, but most  is definitely over my head at least in terms of practical application. Great post though indeed :smile:


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Edited by Psilotyl (12/10/19 10:52 AM)

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Re: Increasing alkaloid levels [Re: Psilotyl] * 1
    #26391885 - 12/20/19 04:37 AM (4 years, 2 months ago)

Sure, ofc thats possible with the manure. I just suspect it can be tricky to figure out how without quantitative experimentation.

As a side note, potency can apparently be offset by yield so there can be a sort of unfairness if you don't consider basically the total alkaloid yield or potency x yield.
Because signs point to mutants such as PE being more potent from growing slower. I think this makes sense if there is disparity in metabolism and development. If alkaloids are produced at a normal rate but the mushrooms grow at a slow rate they would effectively build up more for potency.

I'll think more on the original question of the indole supplementation. Possibly the closer you supplement to psilocin in the biosynthesis pathway the better.
Interesting though.. with a cheap indole supplementation fair question whether it could become worth it to try and compensate for what otherwise seems like an inefficient way to increase alkaloid levels.
Maybe thats the answer to the manure though: if it doesn't just contain simply a source of nitrogen but precursor amino acids or indoles closer to psilocin than having to start from scratch.

Supplementing with tryptophan may be exceedingly inefficient because it can be utilized for protein synthesis, although i guess there would be an excess. It really depends on how creatively it can be utilized. Also depends on which steps in the biosynthesis of psilocin are reversible / how the equilibria lie. idk such details.

Edited by Solipsis (12/20/19 04:46 AM)

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Re: Increasing alkaloid levels [Re: Solipsis]
    #26393694 - 12/21/19 08:25 AM (4 years, 2 months ago)

That’s an interesting thought about PE.

I’ve wondered also when to supplement. Probably during the pasteurization process into sub is the obvious go, as I’m fairly positive substances getting thrown in with grain and going through sterilization would be rendered inactive.

Not really supplementation, but those Shulgin stories are really tantalizing...has me thinking about trying adding some 4-HO tryptamine, close to psilocin like you said, or tryptamine that is readily “4-HO”ed by the mushroom (supposedly) like DPT, DET, MET. They are fairly easy and cheap to get. If I have DMT, it’s getting used for something else (lol). But again like you said, even if at the end of the day the results blow your mind and it works, quantifying what happened and sharing it is an entirely different and difficult endeavor. But at least one would “know”, if the effect was significant enough to warrant assurance on the part of the consumer that something unique had indeed happened. And that would be cool. .


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Re: Increasing alkaloid levels [Re: Psilotyl] * 1
    #26393731 - 12/21/19 08:57 AM (4 years, 2 months ago)

Quote:

Psilotyl said:
That’s an interesting thought about PE.

I’ve wondered also when to supplement. Probably during the pasteurization process into sub is the obvious go, as I’m fairly positive substances getting thrown in with grain and going through sterilization would be rendered inactive.

Not really supplementation, but those Shulgin stories are really tantalizing...has me thinking about trying adding some 4-HO tryptamine, close to psilocin like you said, or tryptamine that is readily “4-HO”ed by the mushroom (supposedly) like DPT, DET, MET. They are fairly easy and cheap to get. If I have DMT, it’s getting used for something else (lol). But again like you said, even if at the end of the day the results blow your mind and it works, quantifying what happened and sharing it is an entirely different and difficult endeavor. But at least one would “know”, if the effect was significant enough to warrant assurance on the part of the consumer that something unique had indeed happened. And that would be cool. .




I would add them after pasteurization (or at the very tail end of the process). I like the idea of feeding the myc tryptamines like DPT, DET, and MET. You could also try out MPT, EPT, MiPT, EiPT, DiPT, and PiPT. I'm not sure how promiscuous the hydroxylase and kinase enzymes are, but it seems reasonable to assume that at least a few of these would work. Please let me know if you go ahead with this! I'd love to see what happens.

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Re: Increasing alkaloid levels [Re: Mycoactive]
    #26393787 - 12/21/19 09:54 AM (4 years, 2 months ago)

I didn't know all the tryptamines, had to look them up...

CLASSIFICATION OF TRYPTAMINES BASED ON THEIR CHEMICAL STRUCTURES

STRUCTURES

Several classifications have been suggested for these substances. Nichols classified tryptamines in two main groups: -the simple tryptamines, including dimethyltryptamine (DMT), and the ergolines (a group of chemical compounds that were originally synthesized from a fungus ergot, among which is the lysergic acid diethylamide (LSD). Fantegrossi divided tryptamines in three subgroups: simple tryptamines, without modification of the indole ring; tryptamines having a modification on the 4-position on the indole ring; tryptamines having a modification on the 5-position. Only substitutions on the 4-5 positions were considered because changes in position 6 or 7 result in reduced hallucinogenic activity


Edited by murderlabz (12/21/19 10:13 AM)

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Re: Increasing alkaloid levels [Re: Mycoactive]
    #26393845 - 12/21/19 10:55 AM (4 years, 2 months ago)

Quote:

Mycoactive said:
Quote:

Psilotyl said:
That’s an interesting thought about PE.

I’ve wondered also when to supplement. Probably during the pasteurization process into sub is the obvious go, as I’m fairly positive substances getting thrown in with grain and going through sterilization would be rendered inactive.

Not really supplementation, but those Shulgin stories are really tantalizing...has me thinking about trying adding some 4-HO tryptamine, close to psilocin like you said, or tryptamine that is readily “4-HO”ed by the mushroom (supposedly) like DPT, DET, MET. They are fairly easy and cheap to get. If I have DMT, it’s getting used for something else (lol). But again like you said, even if at the end of the day the results blow your mind and it works, quantifying what happened and sharing it is an entirely different and difficult endeavor. But at least one would “know”, if the effect was significant enough to warrant assurance on the part of the consumer that something unique had indeed happened. And that would be cool. .




I would add them after pasteurization (or at the very tail end of the process). I like the idea of feeding the myc tryptamines like DPT, DET, and MET. You could also try out MPT, EPT, MiPT, EiPT, DiPT, and PiPT. I'm not sure how promiscuous the hydroxylase and kinase enzymes are, but it seems reasonable to assume that at least a few of these would work. Please let me know if you go ahead with this! I'd love to see what happens.




You could use a 0.22 µm syringe filter to sterilize and inject a solution after autoclaving your sub or medium. That's how they add compounds to media which cannot be heated like that.
Can't they though? As the salt they shouldn't be volatile or sensitive afaik.

I didn't like MiPT but DiPT definitely has 'uses for something else', haha. DPT has such rough edges but 4-HO-DPT appears to be quite amazing and more doable apparently. It just seems all very wasteful but ok. Fascinating if you can find different effects from your mushies that way but imho scientifically i don't see the point in reproducing these results a lot more considering we know too little about the subtleties of actives in mushrooms to begin with at this stage.

DiPT has incredible auditory effects and limited visual effects (although this is all relative and a matter of dosage). Also weirdly while nothing much is seen it can still "feel" very much like being in that DMT world.
Many of those analogues have amazing unique effects (tho not often as sensorically distinct as those auditory effects) even if you shouldn't expect all of them to be such complete packages and classics as the parent compounds like psilocin. The 4-substituted ones are imo probably the best, reasonably paced, physically friendly and still psychedelically adequate imo.

I really hope nobody will overfeed certain toads with trypts to produce novel 5-HO tryptamines...

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Re: Increasing alkaloid levels [Re: Mycoactive]
    #26394052 - 12/21/19 01:42 PM (4 years, 2 months ago)

Quote:

Mycoactive said:
Quote:

Psilotyl said:
That’s an interesting thought about PE

I’ve wondered also when to supplement. Probably during the pasteurization process into sub is the obvious go, as I’m fairly positive substances getting thrown in with grain and going through sterilization would be rendered inactive.

Not really supplementation, but those Shulgin stories are really tantalizing...has me thinking about trying adding some 4-HO tryptamine, close to psilocin like you said, or tryptamine that is readily “4-HO”ed by the mushroom (supposedly) like DPT, DET, MET. They are fairly easy and cheap to get. If I have DMT, it’s getting used for something else (lol). But again like you said, even if at the end of the day the results blow your mind and it works, quantifying what happened and sharing it is an entirely different and difficult endeavor. But at least one would “know”, if the effect was significant enough to warrant assurance on the part of the consumer that something unique had indeed happened. And that would be cool. .




I would add them after pasteurization (or at the very tail end of the process). I like the idea of feeding the myc tryptamines like DPT, DET, and MET. You could also try out MPT, EPT, MiPT, EiPT, DiPT, and PiPT. I'm not sure how promiscuous the hydroxylase and kinase enzymes are, but it seems reasonable to assume that at least a few of these would work. Please let me know if you go ahead with this! I'd love to see what happens.




Word. These all (in theory, as up to date as we know it) should readily undergo four hydroxylation. & I am definitely doing this and it’s just a matter of time. The first experiment will likely either be MET or DPT. Probably DPT and I’ll tell you why... I don’t know if you have done either but MET is in the comparable realm of experience to DMT in many ways - which of course is already chemically taled  to our beloved magic mushroom indoles. That does not lend itself to a “subjective” final test as well as something like DPT which is qualitatively different in experience. It is weird. Powerful and haunting and not like DMT at all. I only consider MET really as maybe a good first choice experiment because I feel, on intuition, it would be four hydroxylated ultra easily due to its similarly with DMT. Like...the mushroom really wouldn’t mind lol and totally go for it. But other than that it is a weaker choice. DPT is really something when ingested that might just end up coming through in a way that conveys positive success. In other words, I envision 4-HO-DPT being something that when ingested I would be possibly like “yep...success!” Hope you follow

DiPT as someone just smartly stated is a terrific theoretical one given the whole audio thing that makes it absolutely unique. The problem is that is it harder to find and much more expensive than DPT. But it might be worth it.

In truth and before moving forward I need to do research on four hydroxylated tryptamine, since that is what we will be getting in the end...which is the most “distinct”, I wonder? Can anyone chime in on that? It would be great to base the decision of tryptamine choice on more than just the TiKHAL entries and Erowid vaults!


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Re: Increasing alkaloid levels [Re: Solipsis]
    #26394055 - 12/21/19 01:46 PM (4 years, 2 months ago)

Quote:

Solipsis said:
Quote:

Mycoactive said:
Quote:

Psilotyl said:
That’s an interesting thought about PE.

I’ve wondered also when to supplement. Probably during the pasteurization process into sub is the obvious go, as I’m fairly positive substances getting thrown in with grain and going through sterilization would be rendered inactive.

Not really supplementation, but those Shulgin stories are really tantalizing...has me thinking about trying adding some 4-HO tryptamine, close to psilocin like you said, or tryptamine that is readily “4-HO”ed by the mushroom (supposedly) like DPT, DET, MET. They are fairly easy and cheap to get. If I have DMT, it’s getting used for something else (lol). But again like you said, even if at the end of the day the results blow your mind and it works, quantifying what happened and sharing it is an entirely different and difficult endeavor. But at least one would “know”, if the effect was significant enough to warrant assurance on the part of the consumer that something unique had indeed happened. And that would be cool. .




I would add them after pasteurization (or at the very tail end of the process). I like the idea of feeding the myc tryptamines like DPT, DET, and MET. You could also try out MPT, EPT, MiPT, EiPT, DiPT, and PiPT. I'm not sure how promiscuous the hydroxylase and kinase enzymes are, but it seems reasonable to assume that at least a few of these would work. Please let me know if you go ahead with this! I'd love to see what happens.




You could use a 0.22 µm syringe filter to sterilize and inject a solution after autoclaving your sub or medium. That's how they add compounds to media which cannot be heated like that.
Can't they though? As the salt they shouldn't be volatile or sensitive afaik.

I didn't like MiPT but DiPT definitely has 'uses for something else', haha. DPT has such rough edges but 4-HO-DPT appears to be quite amazing and more doable apparently. It just seems all very wasteful but ok. Fascinating if you can find different effects from your mushies that way but imho scientifically i don't see the point in reproducing these results a lot more considering we know too little about the subtleties of actives in mushrooms to begin with at this stage.

DiPT has incredible auditory effects and limited visual effects (although this is all relative and a matter of dosage). Also weirdly while nothing much is seen it can still "feel" very much like being in that DMT world.
Many of those analogues have amazing unique effects (tho not often as sensorically distinct as those auditory effects) even if you shouldn't expect all of them to be such complete packages and classics as the parent compounds like psilocin. The 4-substituted ones are imo probably the best, reasonably paced, physically friendly and still psychedelically adequate imo.

I really hope nobody will overfeed certain toads with trypts to produce novel 5-HO tryptamines...




Good info and experience here. I have never done DiPT but am eager too. I honestly just can’t seem to find it anywhere that it’s 5x expensive than the other research chemicals. Regardless it seems like a very good potential choice for experimenting with in the way I have outlined. I’m a musician and would also really just like to try it


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Re: Increasing alkaloid levels [Re: murderlabz]
    #26394074 - 12/21/19 01:57 PM (4 years, 2 months ago)

Quote:

murderlabz said:
I didn't know all the tryptamines, had to look them up...

CLASSIFICATION OF TRYPTAMINES BASED ON THEIR CHEMICAL STRUCTURES

STRUCTURES

Several classifications have been suggested for these substances. Nichols classified tryptamines in two main groups: -the simple tryptamines, including dimethyltryptamine (DMT), and the ergolines (a group of chemical compounds that were originally synthesized from a fungus ergot, among which is the lysergic acid diethylamide (LSD). Fantegrossi divided tryptamines in three subgroups: simple tryptamines, without modification of the indole ring; tryptamines having a modification on the 4-position on the indole ring; tryptamines having a modification on the 5-position. Only substitutions on the 4-5 positions were considered because changes in position 6 or 7 result in reduced hallucinogenic activity






Word. In order for this experiment to work best we would be working the class to the left, there are a lot of them. The result would belong to the middle class. AET/ AMT is a no go way too much going on there with those guys. They have ultralong lasting effects if I’m not mistaken. V Good info & very much sets the “classes” straight or sum such thang. Dang I’m getting excited about this. I could realistically do this as soon as next month possibly. Two tubs of same strain, one control and one supplemented.


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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26395059 - 12/22/19 07:07 AM (4 years, 2 months ago)

Quote:

Psilotyl said:
Word. In order for this experiment to work best we would be working the class to the left, there are a lot of them. The result would belong to the middle class. AET/ AMT is a no go way too much going on there with those guys. They have ultralong lasting effects if I’m not mistaken. V Good info & very much sets the “classes” straight or sum such thang. Dang I’m getting excited about this. I could realistically do this as soon as next month possibly. Two tubs of same strain, one control and one supplemented.




One of the most interesting aspects of this experiment is its unpredictability. If you do manage to get a host of 4-HO and/or 4-PO tryptamines, these will be formed alongside the fungi's endogenous tryptamines (psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, etc.). Their effects in combination with each other are almost entirely unpredictable.

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Re: Increasing alkaloid levels [Re: Mycoactive] * 1
    #26395070 - 12/22/19 07:22 AM (4 years, 2 months ago)

4-HO tryptamines can be bought pure as 'research chemicals'. If one knows how to do TLC analysis it could be interesting to track these 4-hydroxylated being produced with a reference sample.

But since it has already been demonstrated (idk about peer review / reproduction of results) maybe it's nicer to try and 4-hydroxylate something like 5-MeO-tryptamines though i believe this doesn't work.

And yes, while aMT is a wonderful substance and anti-depressant, don't try to 4-hydroxylate it. It may also degrade and smell terribly in the process.

[quote=tihkal]The 4-hydroxy analogue of αMT has been looked at in human subjects. It is reported to be markedly visual in its effects, with some subjects reporting dizziness and a depressed feeling. There were, however, several toxic signs at doses of 15 to 20 milligrams orally, including abdominal pain, tachycardia, increased blood pressure and, with several people, headache and diarrhea.




Basically we seem to be confined to the unsubstituted trypts on the left anyway for candidates, or otherwise lemme think about more unusual derivatives of tryptamines.

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Re: Increasing alkaloid levels [Re: Mycoactive]
    #26395640 - 12/22/19 01:38 PM (4 years, 2 months ago)

I did some
Quote:

Solipsis said:
4-HO tryptamines can be bought pure as 'research chemicals'. If one knows how to do TLC analysis it could be interesting to track these 4-hydroxylated being produced with a reference sample.

But since it has already been demonstrated (idk about peer review / reproduction of results) maybe it's nicer to try and 4-hydroxylate something like 5-MeO-tryptamines though i believe this doesn't work.

And yes, while aMT is a wonderful substance and anti-depressant, don't try to 4-hydroxylate it. It may also degrade and smell terribly in the process.

[quote=tihkal]The 4-hydroxy analogue of αMT has been looked at in human subjects. It is reported to be markedly visual in its effects, with some subjects reporting dizziness and a depressed feeling. There were, however, several toxic signs at doses of 15 to 20 milligrams orally, including abdominal pain, tachycardia, increased blood pressure and, with several people, headache and diarrhea.




Basically we seem to be confined to the unsubstituted trypts on the left anyway for candidates, or otherwise lemme think about more unusual derivatives of tryptamines.</font>




Yes I think it would be best to let the mushroom add it’s “4-HO” & and possibly “4-HO-PO” signatures (psilocin and psilocybin respectively). If I were to simply add something like 4-HO-DPT or 4-HO-DiPT it doesn’t give much room to see the mushroom “shine” as a chemical vessel. Although TLC analysis, which I think should be conducted regardless, MAY show a subsequent increase in these indoles if added, my hunch is that they wouldn’t...but may show something else...Overall, it is a different experiment altogether than the one I have envisioned. I think substituting these, as well as 5-Meo substances, may be a gateway toward raising the endogenous alkaloid levels of psilocybin and psilocin. That’s just a hunch though, but it seems very possible. So it’s definitely something worth doing at some point. However, at this point anyway, it’s not where I am headed.

I’m interested, initially anyway,  in confirming (or disconfirming) the whole notion of four hydroxylation of tryptamine by the mushroom. It is an amazing notion - that psilocybin mushrooms are potentially a vessel of active chemical change...i.e that when presented with a substance not found in nature, a natural organism could empirically alter it into yet another altogether alien tryptamine, four hydroxylated.

Another thought...the research inspiring this experiment is largely inspired by writings in Shulgin’s TiKHAL, which speaks at good length about the “4-HO” transmutation of ring unsubstituted tryptamine but not specifically of “psilocybin-ized” or “4-HO-PO” counterparts being created by the mushroom. I suspect however both will be present. This raises questions on the nature of psilocybin, specifically how dependent it’s existence or creation is on the presence of psilocin in forming itself in fruit  bodies.

Quote:

Mycoactive said:
Quote:

Psilotyl said:
Word. In order for this experiment to work best we would be working the class to the left, there are a lot of them. The result would belong to the middle class. AET/ AMT is a no go way too much going on there with those guys. They have ultralong lasting effects if I’m not mistaken. V Good info & very much sets the “classes” straight or sum such thang. Dang I’m getting excited about this. I could realistically do this as soon as next month possibly. Two tubs of same strain, one control and one supplemented.




One of the most interesting aspects of this experiment is its unpredictability. If you do manage to get a host of 4-HO and/or 4-PO tryptamines, these will be formed alongside the fungi's endogenous tryptamines (psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, etc.). Their effects in combination with each other are almost entirely unpredictable.




I’m still formulating my overall hypothesis, but what I believe will happen is that the natural or “normal” alkaloid production of the four basic psilocybin mushroom alkaloids will be highly suppressed. I’m sure they still will form to a degree, likely dependent at least in part on how much “alternate” tryptamine is added. This leads to another and very crucial question of quantification of HOW MUCH alternative tryptamine should be added? I.e., how much DPT or DiPT should be put into the substrate, exactly? It’s not an easy one to answer and something I will have to research in depth in order to come to consensus on an proper amount. Since this is largely such uncharted territory, it’s something I will probably in the end largely simply decide myself on some sort of - dare I say - “intuitive level.” It might be worth looking at Gartz’s experiment from the 80’s and to somewhat base the amount after his amounts/supplementation ratio; Though an entirely different experiment altogether, it does theoretically provide some information as to what an effective tryptamine level might be.

For now, I am reading through TiKHAL, Erowid & Reddit reports (with a grain of salt), and scholarly articles. I am finding some interesting information that is steering me further yet toward choosing DPT as my chosen tryptamine. Months ago I took extensive notes on a large amount of tryptamine, both four hydroxylated and not - I am currently trying to find them in my office. The jist is that MANY 4-HO tryptamines are well established in being “quantifiably a lot like psilocybin”, as one user experience entry in TiKHAL on one of these 4-HO tryptamine reports. This is something I am trying to avoid.

Here is something I find very interesting: the TiKHAL entry for 4-HO-DPT extremely short and sparse. Shulgin includes no notes of user experience other than at 20mg there “may be a threshold effect.” Eerily, in his comment afterward, the good chemist seems to almost explicitly encourage experimentation with DPT amongst mushroom enthusiasts.

More to come.


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Edited by Psilotyl (12/22/19 01:54 PM)

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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26395707 - 12/22/19 02:24 PM (4 years, 2 months ago)

I need to find more “scholarly” and scientific peer reviewed articles on this whole thing itself, specifically what Shulgin himself had read (if possible) that made him so certain psilocybin mushrooms were capable of this intriguing 4-substitution. Any links to scientific studies that have been conducted and posted in peer reviewed journals would be much appreciated. I hope to be able to commence with my own experiment sometime in the early new year. Thank you :smile:


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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26395731 - 12/22/19 02:41 PM (4 years, 2 months ago)

Okay so I have determined that this whole idea comes from yet another Gartz study in the 1980’s where he added DET to substrate and mushroom bodies produced 4-HO-DET. All I am finding are dead links to the study though :/ Digging deeper, let’s find this thing :smile:


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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26395831 - 12/22/19 03:37 PM (4 years, 2 months ago)

Good luck and please do.

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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26396000 - 12/22/19 05:20 PM (4 years, 2 months ago)

Quote:

Psilotyl said:
Okay so I have determined that this whole idea comes from yet another Gartz study in the 1980’s where he added DET to substrate and mushroom bodies produced 4-HO-DET. All I am finding are dead links to the study though :/ Digging deeper, let’s find this thing :smile:




Looking.. meanwhile check this out -> http://www.cognitiveliberty.org/ccle1/shulgin/blg/2005/12/4-hydroxy-5-methoxy-nn_07.html


"However there is a very interesting study that took place in Leipzig about 15 years ago. Jochen Gartz, a mushroom explorer whom I know quite well, has done some fascinating studies with Psilocybe species by raising them on solid media containing strange tryptamines that are alien to the mushroom. Apparently the enzymes that are responsible for the 4-hydroxy group of psilocin are indifferent to what it is they choose to 4-hydroxylate. He has taken things like DPT or DIPT and put them in the growth media and the fruiting bodies that came out contain 4-hydroxy-DPT or 4-hydroxy-DIPT instead of psilocin. In fact, he has a patent on the process. These active compounds are made by the mushroom so they really are natural and yet they never have been observed in nature. I'll give you even odds that if you put spores of a psilocybe species on cow droppings loaded with 5-MeO-DMT you would come out with mushrooms containing 4,5-HO-MeO-DMT. This way you avoid a 10 step synthesis by growing a psychoactive mushroom that contains no illegal drug - Dr. Shulgin"

UPDATE: Had to use the wayback machine

https://web.archive.org/web/20120321154717/http://countyourculture.com/2012/02/17/biosynthesis-4-substituted-tryptamine-derivatives/

I was thinking it was another paper,  but looks like its just snips from Biosynthesis of 4-Substituted Tryptamine Derivatives ??

Not sure a bout the patent, these are all I find under his name

https://patents.google.com/?assignee=Jochen+Gartz




Edited by murderlabz (12/22/19 05:38 PM)

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Re: Increasing alkaloid levels [Re: murderlabz]
    #26396558 - 12/22/19 11:49 PM (4 years, 2 months ago)

Thanks for all that. Wayback machine. That could solve my problem with dead link. I keep coming across it being hosted on geocities. The study that is. Will post back soon.


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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26396884 - 12/23/19 07:03 AM (4 years, 2 months ago)

It's amazing but normal that organisms transform chemicals they absorb from their environment, it's metabolism. You are what you eat, you know haha.

Enzymes vary regarding how indifferent they are to their substrate. Sometimes they are (almost) so selective that there are (hardly) analogues possible which an enzyme will also accept.. but at other times they can perform changes on a wide range of compounds.
They kinda grab onto molecules with more or less complementary matching areas which can fit or not, or attract or repell. The 5-MeO is i believe thought to just not fit for a tryptamine to be 4-hydroxylated, the 5-MeO gets in the way probably in terms of space ("sterically") or electrostatic effects, doubtful to play as big a role in this case.

Ultimately its a challenge to subjectively tell differences between psychedelics, sometimes you think you know but in a double blind situation it can be suddenly surprisingly hard.
A change in duration of effects could be interesting..

Something else kinda fascinating is basically blocking effects by getting the hydroxylation done on a tryptamine which yields a basically inactive analogue. If there is great competition in the metabolism you could overwhelm most of the psilocin being produced and essentially make inactive cubes haha.
A worthless idea you might think haha, almost but not entirely. :wink:

Anyway it may be good to get as removed from the regular DMT substitution as possible and go with something like DALT or even DBT.

Also interesting seems to maybe attach a certain marker molecule to a tryptamine to easily tell it apart during analysis. There is a bunch of variations on the experiment. idk if i can get the full study/paper.

edit:

Quote:

TROXLER et al. (1 959) synthesized the two ethyl-analogs PT and HT of psilocybin and psilocin as well as many other tryptamine derivatives. These substances were not found in
any naturally growing mushrooms. The reported biotransformation of DT to PT and HT
is the first evidence of a directed biosynthesis of such substances in fungi. In comparison
with the levels of psilocybin and psilocin in naturally growing fruiting bodies (CHRISTIANSEN
and RASMUSSEN 1982, KOIKE etal. 1981, GARTZ 1987, SEME~IEVA etaf. 1986, WURST
et al. 1984), the amounts of PT and HT are higher in the cultivated fruiting bodies of P.
cubensis.
By performing quantitative analysis of the indole alkaloid levels in laboratory-grown
P. cubensis, large variations among different flushes were also found (GAKTZ 1987). It is
interesting to note that the size of the mushrooms was affected by the addition of DT to
the nutrient media. As shown in earlier investigations, cultures on malt agar and rice substrate also produced mushrooms in various amounts and with different appearance (GAKTZ
1987).
Baeocystin was first detected by LEUNG and PAUL (1968), who isolated this incompletely methylated counterpart of psilocybin from Psilocybe baeocystis SINGER and SMITH
grown in submerged culture. The alkaloid was subsequently detected in various collections
352 J. GARTZ
of P. semilanceata (GARTZ 1985a, CHRISTIANSEN and RASMUSSEN 1982, REPKE and LESLIE
1977, VANHAELEN-FASTRE and VANHAELEN 1984).
This investigation shows that even baeocystin as possible precursor in the psilocybin
biosynthesis occurs in significant amounts in Psilocybe species. The high amount of baeocystin as product of the biotransformation of MT indicates a high hydroxylation and phosphorylation capacity of the surface culture of P. semilanceata but a Low ability in methylation of tryptamine derivatives.




Biotransformation of tryptamine derivatives
in mycelial cultures of Psilocybe’
JOCHEN GARTZ
(Received 14 September 1988/Accepted 25 October 1988)  </font></font>

Edited by Solipsis (12/23/19 07:14 AM)

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OfflineMycoactive
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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26397073 - 12/23/19 09:34 AM (4 years, 2 months ago)

Quote:

Psilotyl said:
Okay so I have determined that this whole idea comes from yet another Gartz study in the 1980’s where he added DET to substrate and mushroom bodies produced 4-HO-DET. All I am finding are dead links to the study though :/ Digging deeper, let’s find this thing :smile:




Here's the paper. Seems like the hydroxylase is more promiscuous than the kinase, which is fine as far as psychoactivity goes, although the stability of these 4-HO derivatives will certainly be lower than their phosphorylated counterparts.

Anecdotally, 4-HO-MET is qualitatively  different than 4-HO-DMT. I imagine the same is true for most if not all substituted 4-HO tryptamines.

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InvisiblePsilotyl
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Re: Increasing alkaloid levels [Re: Mycoactive]
    #26401953 - 12/26/19 06:06 PM (4 years, 2 months ago)

Jesus that was an inelegant, confusing read. I’m also not sure that is the droid I’m looking for. I.e. no mention of DET or MET and apparently there is a study out there he did specific to those. I’m a little disappointed in Gartz in general. He apparently did so much wierd and cool stuff in Leipzig, and yet documents of it all are scarce. He doesn’t seem like he was particularly concerned with posterity or passing this stuff onward as much as he was experimenting for his own sake. Idk.


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‎שלום וְאור | PEACE & LIGHT

Edited by Psilotyl (12/26/19 06:12 PM)

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Invisiblemurderlabz
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Re: Increasing alkaloid levels [Re: Psilotyl]
    #26402355 - 12/27/19 01:08 AM (4 years, 2 months ago)

Quote:

Psilotyl said:
I’m a little disappointed in Gartz in general. He apparently did so much wierd and cool stuff in Leipzig, and yet documents of it all are scarce. He doesn’t seem like he was particularly concerned with posterity or passing this stuff onward as much as he was experimenting for his own sake. Idk.




Lookup Prof. Dr. Dirk Hoffmeister  he's been doing some killer research related to psilocybin. Too bad it's going to be for big pharma...

Edited by murderlabz (12/27/19 01:10 AM)

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InvisiblePsilotyl
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Re: Increasing alkaloid levels [Re: murderlabz]
    #26405138 - 12/28/19 05:53 PM (4 years, 2 months ago)

Thanks murderlabz 🙏 will do.


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‎שלום וְאור | PEACE & LIGHT

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OfflineHolyBolete
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Re: Increasing alkaloid levels [Re: murderlabz]
    #26410643 - 01/01/20 09:08 AM (4 years, 2 months ago)

Quote:

murderlabz said:
Quote:

Psilotyl said:
I’m a little disappointed in Gartz in general. He apparently did so much wierd and cool stuff in Leipzig, and yet documents of it all are scarce. He doesn’t seem like he was particularly concerned with posterity or passing this stuff onward as much as he was experimenting for his own sake. Idk.




Lookup Prof. Dr. Dirk Hoffmeister  he's been doing some killer research related to psilocybin. Too bad it's going to be for big pharma...



Thanks for exposing me to this guy.

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InvisibleThomas Envisio
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Re: Increasing alkaloid levels [Re: HolyBolete]
    #28161047 - 01/28/23 09:26 AM (1 year, 1 month ago)

Not to bump an old thread, but I thought there might be something worth noting here. This may sound like a headache, so if you're not in the mood for research, my advice would be to NOT read this next post until you ARE in the right mood. Reading this post may take multiple hours depending on the level of familiarity you have with these topics.

First, allow me to very briefly go over some of the chemicals detected in psilocybin mushrooms, along with some potentially mind-altering notes.

You've all probably read tons of literature on the following compounds, which I will capitalize below, rather than leaving them un-capitalized:

1. Psilocybin
2. Psilocin
3. Baeocystin (Baeocystine)* = misnomer
4. Nor-Baeocystin* = misnomer
5. Aeruginascin
6. Nor-Psilocin

* Note: According to author Paul Stamets, the chemical currently known as baeocystin (baeocystine) was incorrectly named. A misnomer occurred as a result of a mis-identified collection of what was actually Psilocybe cyanescens (and not Psilocybe baeocystis, as was originally published). He proposed cyanescin as the new chemical name, and hinted it could be studied for human therapeutic potential. This would also cause nor-baeocystin to be called nor-cyanescin. See this and fast forward to about 58 minutes through.

See also the important paper titled, Injury-Triggered Blueing Reactions of Psilocybe Magic Mushrooms. Substrates designed to increase psilocybin may also play a significant role in potency and chemical profiles. The third harvested flush of some strains may also play a significant role. As mentioned by others, genetics appear to be the ultimate pathway for altering potency. This requires effort, stamina, and lab gear.

Psilocybin mushrooms also all appear to develop significant quantities of various MAOIs (MonoAmine Oxidase Inhibitors). See this.

See the book called Psychedelics Reimagined by Thomas Lyttle. It includes a section/chapter titled, Psychedelic Monographs and Essays, Volume Six (1993). In it is a page or more about adding dextrose to the substrate (rye in this case) to achieve higher psilocybin content.

However, one mycologist wrote on this site many years ago that this was not the right way to go about it. This particular mycologist said,

Quote:

"I don't see why dextrose would increase psilocybin content. If you really want to increase psilocybin content, add Diammonium Phosphate and non-fat powdered milk to the substrate.  Diammonium Phosphate has the phosphorus needed to make psilocybin, and non-fat powdered milk has the tryptamines."




However, it seems adding tryptamine may also increase psilocybin and psilocin levels.

Jochen Gartz's recipe, as mentioned in previous comments, reciprocally calls for 1/2 pint PF Tek jars: 1/2 - 2/3 cup of vermiculite + 1/8 cup of brown rice powder and (45 milliliters of water with .1 grams of Tryptamine HCL added). Any more than .1 g will cause poorly formed, dwarf-like specimens. However, as experiments in this thread have mentioned, this recipe is now strongly under scrutiny.

As a final stimulating note here, all of the notes provided here require lab gear to confirm these notes or disprove these notes. That lab gear will almost certainly require Liquid Chromatography/Mass Spectrometry. This requires more effort than all members of The Shroomery appear to be willing to put in on this interesting topic. I see a lot of under study, and a massive amount of laziness regarding this topic. If there are literally about 200 species of fungi confirmed for the presence of one or more chemicals relevant to this post, that means someone - or a team - needs to test each species of each relevant genus. Perhaps the person on this resource could lead such an effort.

More For Those Willing To Push The Envelope Of Science:
https://www.agilent.com/cs/library/support/documents/a05296.pdf
https://chemrxiv.org/engage/api-gateway/chemrxiv/assets/orp/resource/item/617755bc913a74cab06a8a2d/original/rapid-quantification-of-psilocybin-with-reversed-phase-hplc-and-single-wavelength-detection.pdf

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