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OfflineGiggle_Grower
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Re: Potency Project [Re: libertaire]
    #12461677 - 04/26/10 07:12 PM (14 years, 8 months ago)

id like to help with this project, tho im not very experienced. im pretty sure experience is what u need for the technical stuff, but if i can do anything else ill try to help. anything for this community


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Re: Potency Project [Re: libertaire]
    #12462518 - 04/26/10 10:24 PM (14 years, 8 months ago)

Quote:

libertaire said:
So who's game for a revival of this?  Shea25 posted this recently, looks pretty legit:

http://www.thenook.org/nooki/index.php?title=Psilocin_Extraction_triptamine

It's an ab extraction, and the end result gets crystals.  The writer of the tek says that it is 70% efficient, which is pretty damn efficient if you ask me.

I intend to try this on the OI and OR I just made lc's of once I have fruits from them, and then later down the line try it on PE whenever that grow materializes, which probably won't be for at least a few months.  Anyone have any fruit on hand they'd like to try this on?  Everything is available at your local grocery store or hardware store. 

So whadya think, are we back in business?  Anyone wanna try this out?




The recipe I posted is already scientifically proven to work and be around 93%-96% efficient.  Why would I want to downgrade to a recipe thats less efficient than the one I already posted?

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OfflineLogicaL ChaosM
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Re: Potency Project [Re: Cloneufc]
    #12462549 - 04/26/10 10:31 PM (14 years, 8 months ago)

I think that calls for a.....



So Cloneufc, still getting that GC/MS machine? When ya do, will you analyze my shrooms for a given amount of cash money....

I really wanna know my Shrooms.
~ LogicaL Chaos ~

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OfflineCloneufc
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Re: Potency Project [Re: LogicaL Chaos]
    #12462592 - 04/26/10 10:41 PM (14 years, 8 months ago)

All I need is an Ultra sonic cleaner. A US jewelery cleaner should work. They are pretty cheap and dont cost much. The recipe calls for Methanol. You can get methanol at any Autozone,checkers ect. The centrifuge is the expensive part but it may not be needed. I found a couple centrifuge's for about $100. The dont go as high as the recipe calls for. A proven 93%-96% is great though.

I am still going to get a GC/MS machine. The extraction should be easy. I will test penis envy against other cubensis sub-strains. Im probably going to need a little help using it. I read what I could on how to use it. No, Im not going to be taking samples from anyone. I have plenty to work with. My main goal is to make or isolate very potent species and sub-strains.

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OfflineLogicaL ChaosM
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Re: Potency Project [Re: Cloneufc]
    #12462775 - 04/26/10 11:17 PM (14 years, 8 months ago)

So, do you have any of those parts: the Ultra-sonic cleaner, centrifuge, or the GC/MS Machine?

I guess I can never convince you to take our samples, but if you post some of your potency results from the PE and regular cubes, that would help this project out TREMENDOUSLY!

It would be biased because of the really small sample size, but its the only objective way to do it. Plus, when you make your own super-PE strain with crazy potency, you can post the graphical results compared to a normal cube from your GC/MS machine and use that for advertising your new strain. I bet people would be all over getting spore prints.

And you bred the mushrooms to make the spores sterile so other people can't grow them and steal your business! That's job security, my friend.

Hope you Get that Equipment
~ LogicaL Chaos ~

Edited by LogicaL Chaos (04/26/10 11:25 PM)

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OfflineGiggle_Grower
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Re: Potency Project [Re: LogicaL Chaos] * 1
    #12462802 - 04/26/10 11:20 PM (14 years, 8 months ago)

i highly doubt that that is Cloneufc intent. just my 2cents


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Re: Potency Project [Re: Cloneufc]
    #12463829 - 04/27/10 06:40 AM (14 years, 8 months ago)

Quote:

Cloneufc said:
Quote:

libertaire said:
So who's game for a revival of this?  Shea25 posted this recently, looks pretty legit:

http://www.thenook.org/nooki/index.php?title=Psilocin_Extraction_triptamine

It's an ab extraction, and the end result gets crystals.  The writer of the tek says that it is 70% efficient, which is pretty damn efficient if you ask me.

I intend to try this on the OI and OR I just made lc's of once I have fruits from them, and then later down the line try it on PE whenever that grow materializes, which probably won't be for at least a few months.  Anyone have any fruit on hand they'd like to try this on?  Everything is available at your local grocery store or hardware store. 

So whadya think, are we back in business?  Anyone wanna try this out?




The recipe I posted is already scientifically proven to work and be around 93%-96% efficient.  Why would I want to downgrade to a recipe thats less efficient than the one I already posted?




I'm glad that you understand the scientific jargon that you posted and have access to all of the materials, and that's great for you, but for people who aren't organic chemists who have access to a legitimate laboratory, the link I posted will make participating in this project a possibility.

I do hope you'll post your results when you get around to it though, you definitely know what's up.

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Re: Potency Project [Re: libertaire]
    #12463833 - 04/27/10 06:42 AM (14 years, 8 months ago)

As long as each person uses the same method for their samples, relative outcomes will be similar.


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OfflineCloneufc
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Re: Potency Project [Re: Doc_T]
    #12464357 - 04/27/10 09:52 AM (14 years, 8 months ago)

Quote:

Doc_T said:
As long as each person uses the same method for their samples, relative outcomes will be similar.




Thats true to an extent but we already have graphs to go by. If the extraction method isnt similar to the percentage of the active chemical its gonna be lower and mess up the results. The samples were between 0.1% to 2.8% or so.

Either way will work if your trying to make a more potent substrain, the results will be based on the highest number or percentage, not on efficiency.

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Re: Potency Project [Re: Cloneufc]
    #12464845 - 04/27/10 11:28 AM (14 years, 8 months ago)

I'm still waiting on more fruits for experimentation...


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Re: Potency Project [Re: NoOneKnowsHowToAct]
    #12467963 - 04/27/10 07:29 PM (14 years, 8 months ago)

Quote:

NoOneKnowsHowToAct said:
I'm still waiting on more fruits for experimentation...




:wave:


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Re: Potency Project [Re: TacoHerder]
    #16114755 - 04/20/12 02:06 AM (12 years, 8 months ago)

I know this thread is very old, but I wanna try this project.

I had just had a brief moment of clear insight (while stoned, strangely).

What if we did a very low-cost, easy-to-do, simple way to compare potencies of mushrooms. So I had a strike of insight.

1.So, first step is to liquefy the mushrooms first. It has to be easy and simple. Pure water maybe is the best, simplest solvent.

2. Ok, heres the important part. We buy chemical pH testing paper. From what I have read, alkaloids are weak acids on a pH scale. So using this info, it can assumed they will show up on only one side of the pH scale, the weak acid side (between 3-6.5 pH). The closer to the ideal pH from the study, which is 5.3 pH, the stronger the mushroom.


-> Ok, now we have to get the ratios right: need a scale, and a volume measurement, like a measuring cup.

We must standardize the portion of both the amount of completely-dry mushrooms powder and the choose liquid to dissolve the active chemicals in the mushrooms into.

1 gram of mushroom : 10 oz of alcohol. We would have to get the ratio right for the pH test trips to measure the appoperiate concentration. Wait, different concentrations does not effect the pH level, right?

So in that way, its fail safe, I think.

Anyway, the measurement of the potency will be the "brightness" of the color test strips is dependent of the concentration of the base or acid.


Basically, making a mushroom tea, at the right water concentration. And of course boiling the water to extract the water properly.


here's some notes on [Chemical Data on Psilocybin from Erowid.org

pH in water: "pH 5.2 in 50% aq ethanol." This means Psilocybin is a weak acid! Acids are near 0 on the pH scale (0-14). Knowing that its a weak acid in water means we can test it with inexpensive pH testing strips.

I just found pH testing strips for $1 on eBay. If they work, they would be perfect. Heres the link to the $1 pH testing strips.

Ok, I think this will work.



Supplies needed


1. Vinegar (see below)

2. Stove to boil water with shroom powder in it.

3. Coffee grinder (to powder the shrooms)  $15

4. A scale to measure the shrooms to exactly (or close as possible) to 1 gram - $20

5. pH testing strips $1.


About $36 bucks. Some people might have these things already.

Procedure:

1) Grow shrooms.

2) Dry them

3) measure out 1 gram exactly

4) grind up the 1 gram in a coffee grinder

5) Boil the 1 gram of powdered shroom in [some] amount of vinegar (not sure yet, needs experimenting) [According to the Gold chemistry article, vinegar (acetic acid) is a much better solvent]

6) After boiling for 1 hour, take a specific amount of the water, (small amount?) and put some on the testing strip.

7) Compare color to just the boiled water (do steps 5-6 but with no shroom powder).

8) Post color photos online for comparison with other people.



Who wants to do this? This will work. I will even buy the test kit and mail strips to people with letters. I don't mind.

Anyone willing to try it? I will be soon, I will be using a small piece of mycelium I am spawning right now.

I will let you know the results when I get the testing papers.



I will start this project soon.
~ LC

Edited by LogicaL Chaos (04/23/12 11:00 PM)

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OfflineRoseM
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Re: Potency Project [Re: LogicaL Chaos] * 1
    #16114890 - 04/20/12 03:01 AM (12 years, 8 months ago)

:smile:

Best bump ever!


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OfflinePrimalSoup
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Re: Potency Project [Re: LogicaL Chaos] * 1
    #16121690 - 04/21/12 08:25 PM (12 years, 8 months ago)

Quote:

Wait, different concentrations does not effect the pH level, right?




No. Adding the same amount of base or acid to different amounts of solvent does affect pH level.

It's been a long time since I read this thread, but here's a further and dire problems:

To extract psilocin into water you need to acidify the water significantly.  Read Gold's work on measuring potency.  Psilocybin extracts easily into water though.  You would only be able to measure psilocybin content with what you're suggesting, and that only if it were the only non-neutral pH substance that goes into solution from the mushrooms, which I sincerely doubt.  When I make tea it has all kinds of crap in it, plus the actives. :shrug:

Quote:

pH in water: "pH 5.2 in 50% aq ethanol." This means Psilocybin is a weak acid! Acids are near 0 on the pH scale (0-14). Knowing that its a weak acid in water means we can test it with inexpensive pH testing strips.


 

Psilocybin may be a weak acid.  But if it's a weak acid at molar concentrations that means it's an exceedingly weak acid diluted in your extraction solvent - unless you have a method to isolate it specifically (of which I understand there are a few, though I've never bothered to do it).

I have a bottle of pH strips I've had forever, and if y'all want a dozen or two just let me know and you can have 'em for free. :thumbup:  But you might as well get a decent pH meter, they're much more sensitive than reading strips.

Other than the above it sounds fine.  Have fun!

:peace:PS


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OfflineLogicaL ChaosM
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Re: Potency Project [Re: PrimalSoup]
    #16131081 - 04/23/12 10:46 PM (12 years, 8 months ago)

Wow, thats really interesting.

I liked Gold's work, he was on to something. And its very similar to my idea. He had a more advanced version of it.

Here's a quote:

"A simple test for indole-containing compounds and tryptamines is to crush a small piece of mushroom into 1/2 ounce of vodka or ethyl alcohol("denatured alcohol" or the hardware store "shellac thinner" is fine) and mix. Add 3-4 drops of hydrochloric acid(or the hardware store variety called, "muriatic acid") then drop a pine tree shaving into the solution which will turn "cheny red" in the presence of indoles."

He did the test with HCL acid, which seems better but maybe a little harder to do.

I thought about using Everclear, like he mentioned here (ethanol) but I did not want to complicate it too much. But I am afraid pure water will be too weak a solvent that it wont get much psilocybin.

And thanks for the offer, I actually just bought 100 strips for about $3.75. If I knew you were given them away for free,  I would have asked first.

Oh well, small loss.

I am curious to see how this turns out.


I have to know Primal, have you tried the pH papers on your mushrooms before?

-LC

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OfflineLogicaL ChaosM
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Re: Potency Project [Re: LogicaL Chaos]
    #16131132 - 04/23/12 10:55 PM (12 years, 8 months ago)

One more thing.

In the article, it says the best solvent to use is acetic acid, which is basically vinegar for the rest of us.

I will try that test first maybe, then compare the pH papers for both the boiled vinegar and boiled vinegar+mushroom powder. That should give a better result (I hope).


That article is a wealth of helpful knowledge! :thumbup:

Thank you.

- LC

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Re: Potency Project [Re: LogicaL Chaos]
    #16133158 - 04/24/12 12:42 PM (12 years, 8 months ago)

Quote:

I have to know Primal, have you tried the pH papers on your mushrooms before?




No, no reason ever occurred.  But since I acidify my tea about 1:50 with citric acid, it's a lost cause. :lol:

Anyway, pure water easily dissolves psilocybin - hardly touches psilocin though.  Hence the acid.  And any acid will work for this purpose.

I really would love to see a simple repeatable test for indole concentration.  I like the idea of posting color results as well, but to make that work you're gonna need some sort of color balance patch in the image - not a biggie though.  Carry on! :thumbup:

:peace:PS


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Re: Potency Project [Re: PrimalSoup] * 1
    #16133966 - 04/24/12 04:01 PM (12 years, 8 months ago)

Here's my input as a research scientist.

Assays are very difficult things to establish.  Unless you have a proven method and only need a slight modification, you need overwhelming evidence of its efficacy before you can make any claims.  Now, this isn't published research, but you still have to have a fairly high standard before you can say that you've established something.  Most of the time, you're just looking at noise.

For the pH test, you're testing pH, not psilocybin content.  There are hundreds of other compounds in mushrooms that are acidic and likely vary significantly within species.  If you select for mushrooms this way, you'll simply be selecting for higher acidity, not psilocybin.  Additionally, the psilocybin content is not nearly high enough to have an appreciable impact on the overall pH, even with a far more sensitive pH assay such as titration.

This doesn't even address the fact that there may be other compounds in mushrooms that are psychoactive.  While psilocybin alone is sufficient for most or all of the effects of mushrooms, that doesn't rule out the possibility of other compounds affecting the experience.  This may be particularly relevant for perceived subtle differences in strain, species, etc.

Essentially, there is a large burden of proof for establishing a robust assay.  While I really do appreciate enthusiasm and creativity, it is somewhat naive to think that a simple method would have not yet been considered.  I'm not saying inspiration doesn't happen, but I'm sure many smart people have spent some time considering this.

_________________________________________________________

Experimental design must start with your ultimate question or goal: How to assay the psychedelic potency of various mushrooms.  There are two basic routes you can take to do this.  (1) The first is to assay for this directly by analyzing the psychedelic effects.  Obviously, there are far too many confounding factors to get a simple consistent result this way.  You would have to rely heavily upon statistics. (2) The second is to use some proxy for psychedelic effect, namely chemical content.  This introduces a set of assumptions, specifically that psilocybin is solely responsible for the psychedelic effects of mushrooms.

(1) To directly assay is the preferred method because it avoids the assumptions of an indirect assay.  Indirect assays require substantial evidence before they can be accepted as reasonable proxies.

The difficulty in this case lies with the high variance of subjective experience, even with controlled dosage and, to the greatest extent possible, set and setting.  However, people citing difficulty with this method (i.e. subjective trip intensity reports) are likely underestimating the power of statistical methods.  Even in cases of high variance, you can get high statistical power with a potentially manageable N (N=number of measurements).  You would need to use the ANOVA (ANalysis Of VAriance) statistical method, which allows you to establish that two data sets are different.  If the effect is non-negligible (say, 20% different, for example) you could likely observe an effect in an N as small as perhaps 40 or 50.  Smaller effects require much greater N values.  Nevertheless, I don't think we care about a variety that is only 5% more potent.  The 20% area is what we're concerned with in the first place.

This assumes that you homogenize a sample, dose accurately, use the same subject, blind the subject (they don't know what they're getting), and control the conditions as best you can (likely indoors, same time of day, day of week, etc).  If you start to use multiple subjects or different conditions, you can compensate, but requires more advanced statistical methods and higher N.  Obviously, you would need to find a very committed subject to do this in the ideal condition.  However, at one session a week, you could see where the results are heading after about 20 weeks.  Essentially, you'd look to see if you p-value was trending down.  Again, I think if someone did this for a year, it might give you a high enough N, but may also require controlling for age/loss of potency (fairly trivial to do so).

All this effort would allow you to conclusively say that one variety was different than another, but does not give an absolute value for potency, so each comparison needs to be done in a pairwise manner.  Assay multiple varieties would require a more elaborate experiment, but ANOVA would still make it possible.

If I were to do this, I would pick varieties that are widely considered to be different, likely different species.  Then, my starting point would be 20 volunteers, once a week, over 8 weeks.  That would give you a good bit of data to play around with, and you could decide whether the experiment works or not.  Anybody doing this needs a good background in statistics and experimental design.

(2) Measuring psilocybin is likely a good method.  Like I've mentioned before, there are assumptions in using this to measure mushroom 'potency', but there is enough evidence to conclude that, at least, this is informative.  It may not give you the whole story, but likely gives you the majority.

Doing this requires chromatography, no question.  Every publication I could find used GC or LC, mostly LC.  This obviously requires a lab and a technician.  Even if you get the equipment, you'll have to go through the same learning curve that every experimental scientist goes through.  Troubleshooting...troubleshooting everywhere.  Science is not easy.  It's not impossible, but I am saying that you're better off trying to find someone who's willing to do it for you.

If there was a better/easier way to do this, I would imagine someone would have found it by now.  There is sufficient interest in this question that I would imagine most of the plausible stains, reagents, and extractions have been explored.  If you have a thorough knowledge of chemistry, perhaps you could try to develop a colorimetric assay, but that would still require lots of validation with LC.

Simply weighing an extract, regardless of purity, would not give you sufficient sensitivity to make useful comparisons.  You might detect wildly different contents, but what's the point in that?

____________________________________________________________

I think the statistical method is far more practical for amateur experimentation.  A reasonably dedicated and thorough investigator could easily acquire the statistical knowledge and design an appropriate experiment.  The results won't tell you all that you would like to know(again, you can only make comparisons, not get absolute values)and would require significant effort and discipline.  However, I think it is achievable.

The holy grail, so to speak, would be a quick assay that would allow you to select for potency over generations.  Even with a simple assay, this would take years of work, but would allow you to get results analogous to what cannabis breeders have achieved.  Come to think of it, most dispensaries assay their plants.  They might have the tools and motivation to help someone out.  If anyone knows if they use GC or LC for this, please share.  I bet a motivated individual would find a willing partner if this were the case.

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Re: Potency Project [Re: shroomtomb]
    #16134238 - 04/24/12 05:03 PM (12 years, 8 months ago)

Awesome bump to an awesome thread!
If I understand what you are saying, I believe you've come to the same conclusions we came to when this thread was current, but you've addressed it with more rigor.
I couldn't get my head around an analytical method, but I developed a poll which gave results I thought were interesting, if not definitive.
I'd enjoy reading your thoughts on the poll, if you're in a mood to comment. :thumbup:


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Re: Potency Project [Re: shroomtomb]
    #16138385 - 04/25/12 03:21 PM (12 years, 8 months ago)

Best first post ever.  +5

:peace:PS


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* potency and temperature yogithehoneybear 5,670 15 04/02/03 06:33 PM
by Smiley_Riley

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