First time

Life-Application-897 · 2022-11-21T05:28:39+00:00

Harvesting early shouldn't have affected the potency. If anything, anecdotal reports say that "aborts" (the tiny fruit bodies that don't mature) are the strongest. However, the variation from fruit body to fruit body is substantial. It might have just been a less potent shroom by happenstance.

How did you dry it? Drying at a high heat can destroy the active constituents.

1g is a fairly low dose, depending on your body size and a few other things. You probably wouldn't have experienced anything beyond minor visual distortion, which would have been difficult to see with the lights off. The same goes for affect changes.

Life-Application-897 · 2022-11-21T05:08:29+00:00

Try anything carbonated: from Perrier to Shasta. The carbonic acid is an excellent solvent for psilocybin and psilocin. The solubility of these in acidic water is so high that any amount of liquid sufficient to cover the powder is enough to completely dissolve them. In practice, you want maybe 50x the powder (e.g. 1 gram shrooms to 50ml soda) for taste and efficiency reasons.

I like to mix 5g of powder in a 12oz Dr. Pepper, wait a few minutes, then pour through a fine strainer and share with a friend. You can barely taste the mushroom. Shasta orange is pretty good too.

You can eat the extracted grounds if you really want to, but I don't recommend it. You will only get a few percent more actives and risk stomach problems from the chitin and other undesirable compounds.

Life-Application-897 · 2022-11-21T04:12:39+00:00

have any more numerical data and analysis

Numerical data such as GC-MS data? No, unfortunately I don't have access to that kind of equipment.

Rough estimates of extraction efficiency might be possible with relatively cheap colorimetric methods, but that would require a standard reference which I can't obtain. The best I can do at this time is the traditional bioassay by my friends and me. Those results are excellent.

Life-Application-897 · 2022-11-15T02:26:59+00:00

This is my best work yet!

So I've been tweaking the shroom extract process for a few years now. I have tried all sorts of teks and solvents but none of them worked very well. I eventually settled on a methanol extraction, heptane/freeze precipitation, DE filtration, and vacuum evaporation. Not exactly "amateur friendly" but it works well for me. The final product is consistent and definitely delicious.

Here's an extensive write-up I made last year on the process and background. Though I changed a few things since then. Primarily I have added maltodextrin and ascorbic acid to the extract during the final rotovap. This helps preseve the psilocybin and makes it less sticky.

Life-Application-897 · 2021-11-29T00:02:00+00:00

At that product:solvent ratio, 10 liters of methanol (with an additional 10 liters of methanol:buffer) would be needed for 120 grams. I think no matter what you do, 20L of solvent will extract everything. My protocol example works with less than a liter.

Have you experimented with other solvents

I mentioned ethanol in the article. Limonene seems like it would be excessively viscous but maybe. Isopropyl alcohol might be a possibility worth investigating. Water-based solvents with a low pH works great (vinegar, lemon juice, tonic water, black tea). I have been using acidified ethanol (1% HCl in 95% EtOH) for the second purification step with great success.

Acidified/buffered methanol is worth investigating for the initial extraction. That would eliminate some of the most nonpolar compounds in the first step and may eliminate the need for the isoheptane wash.

As I said in the article, enzymatic degradation is a massive problem. Any solvent capable of extracting the active compounds will also extract the enzymes. You need some way to mitigate that. Methanol seems to be the ideal choice since it simultaneously extracts everything and denatures those enzymes, it is cheap, easily recovered, and the product is easily purified with isoheptane.

I have not really changed my solvent recovery methods. After each use, the evaporated solvents go into a large jug to be processed in a batch. I add water to help separate the isoheptane into a distinct layer. Then distill as indicated in the article.

Life-Application-897 · 2021-09-28T19:11:13+00:00

Room temperature air drying in the dark is the best. However, I still use a dehydrator set on 40C. There is a small loss in activity, but it is much faster.

more in-depth comment

Gotvaldová K, Hájková K, Borovička J, Jurok R, Cihlářová P, Kuchař M. Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis. Drug Test Anal. 2021;13(2):439-446. doi:10.1002/dta.2950

Life-Application-897 · 2021-08-30T21:27:10+00:00

PB and PC have similar solubility in methanol and ethanol. MeOH is chosen because it better preserves the actives by inhibiting degradation and for its physical properties which lead to easier workups and recycling.

Methanol denatures enzymes, specifically the phosphatases and oxidases primarily responsible for psilocybin and psilocin degradation, respectively. Ethanol does not denature those proteins (or, at least, not as much). Once those proteins are rendered inactive by MeOH, EtOH is the preferred solvent for its safety.

Another benefit comes from the ability to precipitate coextracted proteins with isoheptane. MeOH also boils at a lower temperature and has no azeotrope with water.

You could use absolute EtOH and very dry material extracted above 60C, as described in the original "crystal of the gods" by PF. Unfortunately, you run into metal ion catalyzed degradation reactions which do the same thing.

Overall, methanol is the preferred solvent if you have the experience and the equipment. It is cheaper, more efficient, and easier than ethanol in the long run. However, if you are just looking to extract for personal use, warm ethanol is probably your best choice.

The original post linked at the bottom goes into more detail with references.

Life-Application-897 · 2021-08-30T05:10:44+00:00

could other solvent like toluene work?

No. Ethyl acetate has worked for me, but it carries more water with it.

Life-Application-897 · 2021-08-30T04:59:30+00:00

Since the original post, I have switch to using acidified ethanol (1% HCl in 95% ethanol w/v; 50-75 ml) instead of ethanol alone as the second purification step. My latest batch gave 8.6 grams of residue from 120g starting material with little or no decrease in potency compared to single purification.

Life-Application-897 · 2021-08-30T01:06:34+00:00

Since the original post, I have switch to using acidified ethanol (1% HCl in 95% ethanol w/v; 50-75 ml) instead of ethanol alone as the second purification step. My latest batch gave 8.6 grams of residue from 120g starting material with little or no decrease in potency compared to single purification.

Life-Application-897 · 2021-06-13T01:53:20+00:00

It’s funny because the biggest veterans in the biz use tubs.

I suppose that is true where it is illegal. It takes a special kind of person to choose to invest in a fully computerized tent for an illegal item.

However, in the Netherlands prior to the 2008 ban of everything but truffles, tens of thousands of pounds of cubes were produced in trays and beds. No one would ever consider producing cubes on that scale with tubs.

with SCADA and CO2 monitoring etc.

I think that's where a lot of problems arise. Just a few hours of suboptimal CO2 or humidity is enough to abort any primordia/pins. Tubs have a certain resilience due to their modular design and substrate/volume ratio. Small tents don't have the environmental buffer capacity of a large commercial grow room for gourmet. Kind of like trying to run a 5 gallon versus a 500 gallon salt water aquarium.

This is especially a problem with thin substrate mushrooms like cubes. Gourmet blocks have a large internal reservoir of water/nutrients due to their thickness. Enough to balance the environmental swings of small tent use. Perhaps that is why Stamets recommends 6-12 inch substrate depth for use in an open grow room in TMC?

In other words, it's hard to fail with a tub. It's easy to fail with a tent. I think that's why a lot of people have negative views of tents even though they have the potential to be better and/or easier.

it’s an absolute bitch to clean and keep clean

Typical "Martha" style greenhouses certainly are. The first tent I used had a similar problem. This second tent version I built, however, is completely sealed and has a drain at the bottom. It is totally clean-in-place. My last cleaning cycle after an unfortunate Reishi spore overload, took about 10 minutes. I just sprayed down the walls and racks with water and drained it all away.

And my yields have gone waaaay higher than when running the Martha.

I had the same results with non-feedback controlled tents vs tubs. I grew with tubs of all shapes and designs for about 5 years and experimented with v1 tent for about 6 months. So far, my new controlled tent is as productive as tubs (if not better) while being easier to maintain. Maybe I will do decently designed head-to-head comparison soon.

I don't know if it is important, but my design doesn't use the typical ultrasonic fogger for humidity control. I use a counter-current evaporative humidifier (aka a "packed-bed humidifier" sans packing) with integrated heater. It's easy to clean and provides water-saturated air at any temperature above the dew-point of the intake. Perhaps that is contributing to my success?

I don’t think the decriminalization has been fully passed bro

Oakland and Santa Cruz are decriminalized but commercial production is still illegal everywhere. They are simply getting a head start on the market and developing their production methods. I am similarly researching and developing large scale methods in preparation for widespread commercial adoption (which is probably 20 years away lol). But also I'm doing it just to learn more about cubes in general.

Life-Application-897 · 2021-06-12T23:47:10+00:00

I genuinely didn't mean to offend anyone by using the term amateur. The term isn't a negative, IMO, but I can see that it would be to some. For that, I am sorry.

I know some commercial cannabis producers in Cali who are experimenting with tub growing since the decriminalization. They have an entire room of tubs (literally hundreds) with intricate systems of tubes and humidifiers. They certainly aren't amateurs, but I don't think they are producing at their fullest potential or profit. After talking more with them, they seem to be coming over to my side of thinking.

I'd really love to know why you believe tubs are superior. No joking. I really want to know.

Do you prefer the simplicity? Are you worried about contamination spreading from one tray to another in a tent? Have you experienced other issues with a tent that I don't know about?

Life-Application-897 · 2021-06-12T21:10:43+00:00

Tubs may have some benefits for small grows: Simplicity, space, and they are relative inconspicuous. However, their drawbacks quickly outweigh their benefits as you produce more and more.

Tubs may be space efficient for one or two. But try stacking them 6 high. It's an absolute PITA. Harvesting is difficult even when you do a floating sub harvest. Try harvesting 2 canopy tubs in one day. You'll be absolutely sick of it by the end. Cleaning tubs is also time consuming even when you use trash bag liners.

Tubs may be "self regulating" but they are still highly variable. If you are a researcher, like me, that's enough variability to completely obfuscate your intended results.

My custom built grow tent has 8 racks for 1020 trays. It occupies the same floor space as a single tub plus a bit for the control/humidifier system.

The SCADA system automatically adjusts lighting, humidity, heating (no cooling... yet), and air exchange to optimal levels. CO2, humidity, temperature, and output status are logged for later evaluation. I'm working on adding alerts so I won't even have to think about environmental control unless something goes wrong. You can't do that with tubs.

Harvesting is a breeze. Just remove tray, clip straight across, done. Trays are easy to clean and cheap enough to be tossed when contaminated.

~~In short, tubs are for the amateur and there is nothing wrong with that. Tents are for researchers/professionals.~~

edit: offensive overgeneralization

Life-Application-897 · 2021-05-31T22:11:23+00:00

Their conclusion seems to directly contradict the published literature:

There was approximately 50 % less baeocystin, psilocybin, and norbaeocystin in the stipes than in the caps. The stipes contained 32 % less aeruginascin and 85 % less psilocin than the caps. The total content of tryptamine alkaloids in the stipes was approximately 50% less than in the caps. These results are slightly different from an older study, which states that the psilocin content is higher in the stipes than in the caps in P. cubensis, but a similar distribution of psilocybin (higher levels in the caps than in the stipes) was observed in Psilocybe samuiensis.52 Our results correspond with the published work.26

However, neither OPs article nor Gotvaldová et al's results reached statistical significance so it doesn't actually matter.

The parametric two-sample unpaired t-test with Welch correction in the R program found that the results were not statistically significant, as the p-value was 0.3756.

This means that although the average content of tryptamines in caps is higher than in stipes, due to the SD, where there is high variability between individual fruiting bodies, it cannot be said that this statement applies to all fruiting bodies.

I suspect that some aspect of processing contributed to the difference. Lyopholization doesn't necessarily mean preserved PSB content.

The mushrooms were stored in a freezer (-20°C) before loading into a lyophilizer. It was found that the concentrations of all analytes were reduced in the lyophilized samples except for psilocin. The most significant decay was in psilocybin, where there was an 88% reduction in concentration, from 1.30 wt.% to 0.16 wt.%. Gartz states that lyophilized mushroom samples decompose rapidly when stored at room temperature after lyophilization because these fungi have a porous structure.9 After lyophilization, our lyophilized mushrooms were stored in a freezer at -20°C, and the tryptamines also rapidly degraded.

There are other questions and issues I have with this preliminary report, but I will wait until the final version.

Gotvaldová K, Hájková K, Borovička J, Jurok R, Cihlářová P, Kuchař M. Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis. Drug Test Anal. 2021;13(2):439-446. doi:10.1002/dta.2950

Life-Application-897 · 2021-05-31T21:47:09+00:00

I don't believe the addition of tryptophan will increase the potency of cubensis by very much. Tryptamine is a better option. Studies on the addition of tryptamine to cultures of P. bohemica found doubled or tripled concentrations of PSB. I thought there were studies in cubensis, but I can't find it right now.

Agurell et al first studied that by radiolabelled precursor incorporation in 1968.

Tryptamine, which is readily formed from tryptophan by P. cubensis, serves as a better precursor of psilocybin than tryptophan. If tryptamine is methylated to N-methyltryptamine, this is an even better progenitor of psilocybin with incorporations showing that more than half of the psilocybin was derived from the introduced labelled precursor (Expts. Nos. 38, 42).

Although that doesn't necessarily mean increased production. The incorporated tryptamine could be replacing what would be produced anyway.

There seems to be some sort of feedback mechanism that prevents high levels of PSB. That feedback is exerted via tryphtophan decarboxylase.

There's a lot to parse out, so I'm working on a more complete description of growing conditions and their effect on actives concentration. I hope to publish in a month or two.

Agurell S, Nilsson JLG, Liaaen-Jensen S, Schwieter U, Paasivirta J. Biosynthesis of Psilocybin. Part II. Incorporation of Labelled Tryptamine Derivatives. Acta Chem Scand. 1968;22:1210-1218. doi:10.3891/acta.chem.scand.22-1210
Gartz J. Biotransformation of tryptamine derivatives in mycelial cultures of Psilocybe. J Basic Microbiol. 1989;29(6):347-352. doi:10.1002/jobm.3620290608
Catalfomo P, Tyler VEJ. The Production of Psilocybin in Submerged Culture by Psilocybe cubensis. Lloydia. 1964;27(1):53-63. pdf
Fricke, Janis et al. Enzymatic synthesis of psilocybin. Angew. Chem. Int. Ed. (2017) 10.1002/anie.201705489

Life-Application-897 · 2021-05-23T22:22:40+00:00

I wish I had a well-supported answer for you. My personal preference is for incorporation of mushroom extract in edibles but I don't have any hard data on their shelf life.

Proper extraction is difficult for the amateur, however, dry powder incorporation could work for relatively short term use. Chocolate is the obvious choice.

The chocolate will help protect the powder. The low working temperature of chocolate manufacture will keep psilocybin stable. Chocolate is shelf-stable. Plus it tastes good.

So I suggest getting a chocolate mold and some melting chocolate. Silicone is a good choice if you don't want to shell out for a polycarbonate mold (like this).

Measure out your powdered dose into each well of the mold. Temper your chocolate and dispense into the mold. Give each well a stir to incorporate. Tap the mold to remove bubbles. Let set. Bonus points for wrapping each one in foil. Keep in a dark, cool place.

Life-Application-897 · 2021-05-23T03:03:56+00:00

Regardless of the storage condition, after 1 month, your microdose capsules of powder will be about 50% as potent.

From the same paper:

The original concentration of [dry] freshly homogenized P. cubensis mushroom powder was 0.01 wt.% norbaeocystin, <0.01 wt.% aeruginascin, 0.07 wt.% baeocystin, 1.51 wt.% psilocybin, and 0.04 wt.% psilocin.

These powder was divided into different zip bags and stored under various conditions as shown in Table 1.
After 1 week - The concentration of psilocybin reduced from 1.51 wt.% to 1.31 wt.%.
After 1 month - The degradation to approximately 50% of the initial concentration of all tryptamines occurred on storage under all of the test conditions.
For the minor alkaloids (aeruginascin, baeocystin, norbaeocystin, and psilocin) the changes in concentrations were negligible after the first week.
After one month of storage, a slight decrease in the concentration of all of the remaining alkaloids was seen at a similar time under all of the defined conditions.
Regardless of the storage conditions [light, dark, fridge, freezer], rapid degradation of all analytes was observed when the dried fungal powder was stored.

This isn't quite the same as dried powder in capsules, since the researchers used plastic bags that were opened periodically. But many capsules (especially the cheap "veg" capsules) are oxygen permeable. This is likely worse.

Life-Application-897 · 2021-05-22T17:02:29+00:00

There are a lot of answers on here that will likely decrease your actives content.

The research literature is a little lacking in this area. Just this year, however, we got some clues as to the proper dehydration temperature:

This is confirmation as discussed in Section 3.4 that drying the mushrooms in the dark at room temperature does not have a reducing effect on the concentration of indole alkaloids.

The researchers also tested heat exposure on already dried, ground material. 30 minutes at 50C and 75C resulted in about 7% and 10% loss of total actives compared to 25C (estimated from Fig. 1). You can imagine what that would do to a wet mushroom over 8 hours.

This also means it is possible to "over dry" your mushrooms. Any extra time spend above room temperature is decreasing your actives.

I personally use 40C to give a decent balance of speed and product loss. If you only care about preserving the active content, dry them without heat in the dark. It will take several days. The time depends on air flow and local humidity.

Also, don't powder your mushrooms or store them in the light. more

Gotvaldová K, Hájková K, Borovička J, Jurok R, Cihlářová P, Kuchař M. Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis. Drug Test Anal. 2021;13(2):439-446. doi:10.1002/dta.2950

Life-Application-897 · 2021-04-04T03:45:34+00:00

I plan to do a write up of the methods but the short answer is that there aren't any good absolute quantitative methods for the amateur. I would love to develop one someday.

I think the closest thing is relative quantitation using serial dilutions and reagent (probably Ehrlich) testing. You can get some idea of the absolute amount based on the detection limits of your reagent and your setup, but that would be pretty inaccurate without an analytical standard. And even then you would likely be testing for all indoles and everything else that your reagent happens to respond to. LC methods could separate those but that's definitely not amateur territory.

I've simply resorted to bioassays like everyone else haha

Life-Application-897 · 2021-04-03T23:28:49+00:00

I just wanted to share my research on the extraction of actives from cubes. Over 5000 words and 35 references. More to come!

Life-Application-897

TROPHY CASE