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Offlineshroomtomb
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Re: Potency Project [Re: Doc_T]
    #16160431 - 04/30/12 01:41 PM (11 years, 10 months ago)

Quote:

Doc_T said:
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:




I think that poll is exactly what we in the science community would call "preliminary data."  These are one-off little experiments and observations that we use to justify further research.  Which really means, we say, "Look at this data!  Give me grant $$$$!"  The problem with the poll is that there are no repeated measurements, no way to standardize the data, and strong sampling bias (e.g. people who care about the potency issue are more likely to comment).  HOWEVER! even with all that, there's a very strong result for the PE.  And you made a valiant effort to control for what variables you could.  If you could weight the data based on experience level and whether they grew both strains, etc., it would be even more interesting.  Nevertheless, a result that strong is compelling, to say the least.

This would be most useful in establishing an assay.  You would develop a methodology, and then establish whether it is sensitive enough to detect a difference between PE and some cubensis of your choosing.  Ideally, it would readily detect this difference, and you could move on to attempting to detect more subtle differences.  You would also have PE as a positive control that you could use to standardize your data.

Obviously, the goal would be develop a method that would be robust and easy enough to use that you could select for potency over generations.  Realistically, I think this would have to be around 4 or 5 experimental sessions per participant, and maybe half a dozen participants.  Any more, and it's simply too much work.

My Thinking

The more I'm thinking about this, the more I think this could be workable.  In particular, I think that if you could optimize conditions to minimize variance, you may be able to do this with just a handful of friends over a month or two, and still be able to compare, say, half a dozen isolates to pick the strongest one.

I think the key to optimizing would be to assay at small doses.  First, this would make it much easier to fill out a simple questionare at multiple points during an experiment.  From that, you could get a nice dose-time-response curve that would greatly help even out the 'noise' from collecting data at one particular point.  In other words, the data you collect could be the sum total of all the data points, possibly with some filtering to throw out outliers.  That way, you should be able to minimize temporary effects of 'waves' of tripping.  Additionally, I think it is much more difficult to quantify, or even describe, effects at higher levels.  With small doses, you can indicate discrete effects such as:

visuals:  [glowing] [tracers] [pincushion/barrel distortion] [breathing]

auditory: [humming] [flanging]

somatosensory: [shroom fingers] [deep muscle relaxation] [altered sense of body size/shape] [vertigo] [dizziness]

emotional: [introspection] [memories] [anxiety]

and so forth.  It would be easy to do a series of different doses with each participant, and generate a personalized quantification system, where certain symptoms correspond to a quantification of intensity.  For example:

At 0.5g dose, subject A experiences symptoms 2,4,5
At 1.0g dose, subject A experiences symptoms 1,2,4,5,8
At 1.5g dose, subject A experiences symptoms 1,2,3,4,5,6,8,9

You could adjust the questionnaire as needed to better approximate what is being experienced, and eventually, you might even notice discrete sets of symptoms that correlate, even between subjects.  At any rate, it would give you something far more robust than, "pick a number from 1 to 10, how hard are you tripping?"

So basically, you print out a bunch of notecards where you can check off various things, feed the data into a spreadsheet, translate the data into a number, and get some sort of dose-time-response curve out of it.  You'd also get some indication of the nature of the effects, not just the intensity.  So you could more definitively say that something is more 'clear and pure' or 'dark and sinister'.  This may be even more interesting when all is said and done.

Just remember that this is really only useful for comparing things across what an individual participant has assayed.  Perhaps when there's a lot of data, you could start seeing obvious differences across 'strains'/species, but that wouldn't be the primary goal.  It would be most useful for comparing different isolates to select for potency over generations.

And the best data would come from maximum control over the experiment.  You would have to grind/homogenize a batch to sample from, precisely dose, ingest in the same way, and try to do similar things during the experiment.  And who knows if the effects of small doses correlate that well with large doses?  I suppose you could test that, but I'm just trying to show that this would need to be done very carefully and thoroughly to get a robust result.

That said, I'm going to print up some notecards and try a few small dose-time-response curves from some blended-up boomers.  I think my method of intake will be tea, as that seems to be the most consistent, and doesn't rely on the state of your digestion as much.  It also makes the assay take less time and material, both ideal qualities.  If I'm intrigued by the results, I'll start up a new post and get community input on making a standardized questionnaire card.

And if anyone is curious, I thought of most of this during the symphony yesterday with some tea with 1.5g of bonus ingredient.  I noticed that it was fairly easy to keep track of the effects over time, with extra attention paid to how variable it was.  I think many time-points will be needed.  At the end of the concert, I felt back to baseline, but sure enough, once I got up to walk back to the subway, I was most assuredly not back to normal.  Nevertheless, the overall experience was fairly consistent and predictable.

Also, thanks PrimalSoup!

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InvisibleDoc_T
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Re: Potency Project [Re: shroomtomb]
    #16160467 - 04/30/12 01:50 PM (11 years, 10 months ago)

Quote:

  If you could weight the data based on experience level and whether they grew both strains, etc., it would be even more interesting. 




Oh, but you can! :smile::thumbup:

At the bottom of the poll are two choices to see results. The second one is the one I linked above.
On that results page, you can click on a result and filter out those responses.
So for example, the seven people who pick Grandmaster as their experience level voted 6-0 for STAL over UTSE.

Thanks for your feedback on the poll, I put a lot of thought into it. Glad it was worthwhile.


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OfflineLogicaL ChaosM
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Re: Potency Project [Re: shroomtomb]
    #16180388 - 05/04/12 04:27 AM (11 years, 10 months ago)

Yes, you are very right about all of that, but my simple yet incomplete theory is that the acidity of the mushroom structure (flesh) is far less acidic than the alkoloids psilocybin and psilocin.

So I did a simple pH experience to provide some "proof" of concept. Its not's cause-effect proof, but its correlational.

PrimalSoup, you make a great point about using a digital pH meter instead of strips. Sadly, Im completely broke right now, so using I am using pH strips for now. Once I get some personal funding, I will set-up the project.


Here's what I did for my pH strip experiment.



First, I wanna demonstrate how pH strips work and how they indicate pH from changes in color.

Without going too much into what pH, I will simply say that every substance has a pH and it usually ranges from 0 - 14 on a scale. 7 is considered neutral, anything below 7 is considered an acid and anything above 7 is a base.

There are specific physical characteristics of acids and bases.

Acids are sour (i.e. vinegar) and bases are bitter and slippery (i.e. soap).

This is my sort of evidence that the mushroom's pH have more strength from the alkaloids than the other materials in the mushroom, because when you eat mushrooms, they subjectively have a sour taste (which makes me sick to my stomach from the taste).


Anyway, heres the demonstration of how pH strips work:

Here are the pH strips



Here are the supplies I used: Jar, white PC paper, sharpie, eye droppers, and pH paper.




The strip when its dry



1st Comparison: Tapwater from the sink




- Note that the color of the dropped strip lies in-between 7 and 8 pH. Like an Olive green color. This is expected as water is neutral (7 pH). The procedure is to take one strip, cut it in half, and drop liquid on one of the pieces of the strip. The other is left dry to is to show color comparison to a control color (no drops).


Next is to try a weak acid, lemon tea. It has many other substances, but the main acidic ingredient is citric acid.



Strip Color Comparison: Pure Lemon Juice




- Note that I dropped on a white PC paper. That is to compare the color of the liquid to the strip with the drop. If they are similar colors, this means that the coloring in the liquid is affecting the color of the strip instead of the pH acidity or basic state of the liquid. In this case, the strip is much darker than the water dripped onu the paper. But that could be because of the mixture of the papers non-white color with the juice color. This leads to the next experiment.

Diluted at 1/4 cup of lemon juice to 3/4 cups water





- There is still some color in the diluted juice, but as you can see, the color of the pH strip is much more green (toward neutral) than orange (toward acidic). This shows that color of the liquid has less of an effect than the reaction of the pH to the liquid, but only when the colored liquid is diluted.

The Vinegar test: A dilute acid (bottle says "5% acidity")



- Vinegar is one of the best substances to test for acidity and sensitivity of the pH papers since it is clear as water. This shows dramatic changes in pH color of the paper just buy the acidity of the vinegar. In the photo, the paper appears bright Orange, which is expected as colors toward the reddish-orange range are acidic. I compared it to the lemon juice, which is a weaker acid in comparison of colors.

Bleach (a base) compared to other tested aqueous substances:


Note how bleach has a green color even thou its basic. Not sure what to make of this. I am thinking that the fact its "regular" makes it much more dilute than maybe "ultra" type bleach. I have not yet tested for a very basic substance, but it really doesnt matter for this experiment since I was testing for acids (alkaloids).


Ok, so the strips have been tested, and they do what they are intended for: change color relative to the pH of the watery substance added to it.


Testing a Mushroom's pH

So now onto the real experiment, the potency of mushrooms.

I took one of my potent wild cyan mushrooms for testing. I am not really trying to see the potency of the mushroom, but rather take a mushroom with a high concentration of alkaloids (a weak acid) and see how it would effect the testing strips.

One Ps. cyans



Vinegar is used as a solvent to grossly extract the active components of the mushroom, as mentioned in a paper at Erowid.org under "Mushroom Potency".
I then boiled the chopped-up mushroom in about 1/3 cup of vinegar for 30 min. I had to add more vinegar as the vinegar slowly boiled away. In a separate pot, I boiled vinegar by itself to have a control sample to compare to.


(by the way, boiling vinegar in an open pot is nasty, I recommend a cover of some sort or glass flasks, if you can get them.)


I skipped some photos but here is the conclusion.


All compared together: boiled vinegar (bottom left), 1 drop of concentrated Ps cyan extraction + 1/4 cup vinegar (top) and vinegar/mushroom boiled to a concentrated drop. 



Conclusions: As seen in the photo, there are differences between the diluted drop of ps. cyans and vinegar vs. plain vinegar. I will be disregarding the concentrated vinegar/ps cyan extraction because the color of the strip is not even on the color pH scale.

If you compare the colors of the diluted cyans/vinegar extraction to vinegar, there is a subtle but present difference in darkness of the orange color. As indicated by the scale, the darker orange, the more its closer to a strong acid. And if you check the top right hand corner, the color spot on the paper is clear. This shows that the diluted solution has a little more acidity than the vinegar. Could this be the acidity from the Ps cyan mushroom? I am not sure, it would need more testing.

These results also show how insensitive the pH paper strips are to presence of the mushroom extract. This suggests a much higher resolution to seperate out the differences in pH. A electronic pH meter could be the answer.


As a disclaimer, I did not follow the most strict, exact, precise techniques. And my extraction is pretty crude anyway. But the colors of the two compared strips show a pretty obvious difference, even if subtle, in color.


There are many things that would need to be standardized for this to be even closely helpful.

Color of light on strips, how to best extract the alkaloids, the amount of solvent to mushrooms, how much dried mushrooms to use, what method to process the mushroom (powder or just cut it up?), boiling/extraction, etc....


Cant wait to get a digital pH meter, once I find a way to make money. I hope the meter is accurate.

Until then, I may do some more mushroom experiments .


Side Note: pH paper is very absorbent!

Perfect for Acid dropping. :lsd:


This pH scaling comparison needs much more refinement, standardization, and even better, isolation of the alkaloids to measure the pH of the substance directly to then compare to mushrooms.

Its cheaper than Mass Spectrometry :shrug:
~ LC

Edited by LogicaL Chaos (05/04/12 04:39 AM)

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OfflinePrimalSoup
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Re: Potency Project [Re: LogicaL Chaos] * 1
    #16187703 - 05/05/12 08:15 PM (11 years, 10 months ago)

You'll want to verify the accuracy of the pH meter if you get one - this is easy enough using a known molar acid and dilution, much like what you've done here.

As an aside I've never dried the pH papers when testing.  I can't imagine why you'd want to go to that trouble - the instructions for my vial of papers say to shake excess water off and then compare directly to the color chart.

:peace:PS

PS a lot of things are cheaper than mass spec... :lol:


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OfflineLogicaL ChaosM
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Re: Potency Project [Re: PrimalSoup]
    #16191903 - 05/06/12 07:45 PM (11 years, 10 months ago)

Definitely, I want to get a digital one, but need to sure its consistent.

I have seen some online for like $9. I hope they are accurate. Theres higher resolution ones with 0.01 pH readings, but they are $40. So thats out for now. $9 for me.

Also, I didn't dry the strips, I left some dry, as in I did not drop any liquid on half the pieces to show a comparison to a dry strip that is completed untouched by liquids.
"Excess water" huh? I better think of a better why to strain them. But with the digital one, this wont make a difference.

Anyway, the experiment will continue soon, hopefully with some better results.


-LC


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