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Talk to me about potency!


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#21 Deleena24

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Posted 14 January 2019 - 02:47 PM

Thank you good sir! This is exactly the type of honest feedback I was looking for. There definitely seems to be two camps on this issue, have you grown any of the apparently 'extra potent' strains like PE?


I have grown PEU recently, but haven't had the chance to test.

What I can say is that they fruit very slowly, which, according to accepted thoughts right now, results in more actives. They also blue more heavily than any other species I have worked with, even with being extra careful not to handle them too much.

PE was bred and inbred over and over, so a strain isolated for specifically for mutations could very well have a gene that makes for more actives production as well. Maybe they blue more bc of more psilocin compared to the average cube, which accounts for people consistently agreeing they're stronger than the usual cube.

This is all speculation of course.

I think what I am getting to is that some strains have been worked on and refined so much that they might have accidentally bred out certain Gene's related to potency. Or mutated strains like PE dont have the gene for regulating the amount of actives, so they produce more.

The more I read about genetics and variance in cubensis the more I am interested.

I mean, look what we did with dogs and wolves. They are 100% the same species, but the variance in how their genes are Expressed is so great the range of its strains or breeds is enormous. They look, act, and behave completely differently, while still being the same species.

This stuff is so interesting to me.

#22 raymycoto

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Posted 14 January 2019 - 02:52 PM

A rehashing the above, but my $0.02

I believe there is 'tolerance' to entheogens, psilocin, dmt, etc, at three levels.

  • Receptor level - The biochemical level. The entheogens as are seratonin receptor agonists, tryptamines, substituted dimethytryptamines, that is. And as such their action at the 5HT2a receptor is quickly down regulated as with many catecholamine receptors.
  • Brain level - their action at the 5HT2a receptor is to provide antagonistic action along other pathways, hence the term "shutting down the default mode network (DMN)". I don't doubt that there is some way that the brain fights back to reestablish these primitive pathways, thus perhaps tolerance may occur or other higher, yet subcognitive levels.
  • Conscious and behavioral level - most of us are all familiar with the factors of set and setting which influence a favorable response to entheogenic treatment. I can't help but think that increasing interval between dosing does not lead to a more meaningful or beneficial setting. We 'need' the therapy more as time goes by but also with time we assimilate what we have learned and implement cognitive and behavioral changes. If we are lucky we have realized some favorable feedback not only in through our own mindful assimilation but perhaps through societal interaction - family or work, for example. Dosing at too short intervals may short cut this important process of assimilation.

Have respect for the power of these truly unique drugs. Wait and savor opportunity to redose. Be grateful that you have both access to them as well as the knowledge and discipline to use them for good. Teach others to do the same.


Edited by raymycoto, 14 January 2019 - 02:57 PM.

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#23 HooKworm

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Posted 14 January 2019 - 03:35 PM

Now this is how a conversation goes. I haven’t tested the waters here yet as to personal opinion , and this is refreshing. Bunch of good fellas in this thread. A lot of good info here as well. I agree with everyone who has commented here , 100%.
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#24 PistolPete13

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Posted 14 January 2019 - 05:30 PM

So how are you actually going to measure the % of each of the 4 known actives?

Do you have the equipment that can do atomic spectroscopy? If you do, where have you been for the last decade!?

Nobody has accurately tested potency in fruits for what, 50 years?

 

So I had to spend a little $$ to get the right equipment to be able to quantify(measure the quantity) rather than just qualify(test for its presence). As stated I am using TLC, and one of first things I had to get were analytical scales that measure down to the milligram so 0.001 of a gram. Years ago that would have been thousands alone, but the price has come down dramatically. So every sample will weigh exactly 100mg, the scales actually have an enclosure over them so that drafts in the room dont throw it off! Then a good quality pipettor is a must, I have one that sucks up exactly 1000ul (microliters) every time (it is adjustable down to the microliter). Then extremely important is a good quality capillary tube, I was not going to get into the nitty gritty here but there is really no other choice than Drummond microcaps if you want professional results. The ones I have get precisely 2ul every time. This way everything is exactly the same every time, except the contents of the sample.

 

Basically you add 100mg of dried powdered sample to a small eppendorf tube, add exactly 1000ul of solvent to the eppendorf with the pipettor, close and shake. Repeat with all the samples, after a predetermined amount of time a tube is opened and small piece of cotton ball is dropped in as a filter and the capillary tube pushes it down and draws up 2ul of sample to be tested.

 

Tryptophan is extremely similar to psilocybin, so a reference solution of a known quantity(say 2%) can be made and run on the plate. To give a ballpark idea of what 2% looks like, in my experience that is all you need, you can scan the plates on a scanner so you get the same size photo every time and compare the sizes.

 

There is free software available now that analyzes the plates for you, you literally take a photo of the plate and it quantifies it by dissecting the code in the photo and getting the color values. It is pretty amazing here is a screenshot of an older one that works quite well;

tlc_analyzer.gif

 

https://www.scienceb..._analyzer.shtml

 

And now I have the chance to do things that actually interest me, I am going to be meticulous!

 

BTW; Might not have to wait very long, the 5++ year old creeper syringe has already come to life! There is a Creeper MS on the way!!!!

20190101_150601_resized.jpg

You can see the spores all clumped together over the years, in the creeper there were heaps in the tip. So instead of squirting a syringe onto a plate I just transferred the clumps. I still had a bit of water come over, hence the condensation and the reason I could not get a great photo.

U.jpg


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#25 PistolPete13

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Posted 14 January 2019 - 05:42 PM

led.jpg

 

We are talking about C, I have the Rf values for a ridiculous amount of possible metabolites of interest using this solvent system. C is 2D chromatography so traditional TLC would have all those compounds at the top bunched together. If I wanted to elucidate the metabolic pathways the plate would have to be developed again on its side to get them to separate (thats why they are not in a straight line above). But without having to do develop it again you can see that you get good separation of psilocin and psilocybin from the rest of the compounds.

 

I was thinking it would be great to test extracts also, to see exactly what that brown tar is for example. And if its actually urea that upsets your stomach?

 


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#26 raymycoto

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Posted 14 January 2019 - 06:10 PM

But you're not really talking about being quantitative, just qualitative?

And this requires some sort of standardized extraction from the fruit. What process would that be?


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#27 Deleena24

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Posted 14 January 2019 - 06:23 PM


So how are you actually going to measure the % of each of the 4 known actives?

Do you have the equipment that can do atomic spectroscopy? If you do, where have you been for the last decade!?

Nobody has accurately tested potency in fruits for what, 50 years?


So I had to spend a little $$ to get the right equipment to be able to quantify(measure the quantity) rather than just qualify(test for its presence). As stated I am using TLC, and one of first things I had to get were analytical scales that measure down to the milligram so 0.001 of a gram. Years ago that would have been thousands alone, but the price has come down dramatically. So every sample will weigh exactly 100mg, the scales actually have an enclosure over them so that drafts in the room dont throw it off! Then a good quality pipettor is a must, I have one that sucks up exactly 1000ul (microliters) every time (it is adjustable down to the microliter). Then extremely important is a good quality capillary tube, I was not going to get into the nitty gritty here but there is really no other choice than Drummond microcaps if you want professional results. The ones I have get precisely 2ul every time. This way everything is exactly the same every time, except the contents of the sample.

Basically you add 100mg of dried powdered sample to a small eppendorf tube, add exactly 1000ul of solvent to the eppendorf with the pipettor, close and shake. Repeat with all the samples, after a predetermined amount of time a tube is opened and small piece of cotton ball is dropped in as a filter and the capillary tube pushes it down and draws up 2ul of sample to be tested.

Tryptophan is extremely similar to psilocybin, so a reference solution of a known quantity(say 2%) can be made and run on the plate. To give a ballpark idea of what 2% looks like, in my experience that is all you need, you can scan the plates on a scanner so you get the same size photo every time and compare the sizes.

There is free software available now that analyzes the plates for you, you literally take a photo of the plate and it quantifies it by dissecting the code in the photo and getting the color values. It is pretty amazing here is a screenshot of an older one that works quite well;
tlc_analyzer.gif

https://www.scienceb..._analyzer.shtml

And now I have the chance to do things that actually interest me, I am going to be meticulous!

BTW; Might not have to wait very long, the 5++ year old creeper syringe has already come to life! There is a Creeper MS on the way!!!!
20190101_150601_resized.jpg
You can see the spores all clumped together over the years, in the creeper there were heaps in the tip. So instead of squirting a syringe onto a plate I just transferred the clumps. I still had a bit of water come over, hence the condensation and the reason I could not get a great photo.
U.jpg

That's amazing! Thank you for doing the work, and buying the equipment. I don't know anything about TLC but I will read up on it so I can better understand the process.

If you're doing what I understood from the explanation, the community has waited a long time for this.
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#28 PistolPete13

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Posted 15 January 2019 - 12:02 AM

So have I



#29 PistolPete13

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Posted 15 January 2019 - 12:14 AM

But you're not really talking about being quantitative, just qualitative?

And this requires some sort of standardized extraction from the fruit. What process would that be?

 

 Sorry I missed this post;

 

I have written the basic process above, I can go into greater detail if you like. The biggest drama quantitatively speaking is getting the weight of the sample, the amount of solvent and amount spotted onto the plate exactly the same everytime. We will not get a number like 0.89% psilocybin, but after making a reference solution of 2% tryptophan and running that we will have a ballpark figure for 2%. And will easily be able to tell which spots are bigger just by eye, I will scan and upload all the plates so the pics are the same ratio and size ect every time, but then running them through this software will give you a figure that can be used relative to other tests. From the website;

 

 

Thin-layer chromatography (TLC) is a widely used method for qualitative analysis to determine the number of components in a mixture, to determine the identity of two substances, or to monitor the progress of a reaction. The more accurate high-performance TLC (HPTLC) is better suited for quantitative analysis. Unfortunately, HPTLC requires expensive equipment that most high schools and colleges cannot afford. If digital photography is combined with regular TLC, it can perform highly improved qualitative analysis as well as make accurate quantitative analysis possible. This novel, digitally enhanced TLC (DE-TLC) is easy to use. A fluorescent TLC plate is illuminated with UV light and a picture of the plate is taken with a digital camera. Then, on a computer, using either TLC Analyzer, the public domain software written for this work, or common photo-editing software, one can quickly produce multi-spectral scans, densitograms, and calibration curves—output previously available only from more expensive equipment or complex procedures. Digitally enhanced TLC is a valuable tool that can be added to every chemist's TLC toolbox. Since this technique is less expensive than other quantitative chromatographic methods, DE-TLC is ideal for high school and college labs.

On this webpage you can download and learn how to use the software that enables DE-TLC:  TLC Analyzer.

https://www.scienceb..._analyzer.shtml

 

I have seen one of these programs in action and it was quite impressive!



#30 raymycoto

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Posted 15 January 2019 - 12:36 AM

Maybe I'm not seeing it. What method / extraction from the fruit do you use to get the 1cc from which to do the modified TLC? That will have to be both a thorough extraction of everything we want to assay and will have to be standardized to come up with any comparisons. That is, (making this up) take 10 grams of fruit, 100 cc solvent, puree, mix, reflux, soxhlet or whatever, concentrate to 10 cc, then do your digital TLC. We are talking about subtle differences so one must arrive at the test solution in a standardized way.

 

I am interested in extraction and this would be a way to test what really works - water, EtOH, MeOH, IPA etc.  Take a large quantity of fruit, extract several ways and quantify. In addition, find out what really works for preservation of the magic and what destroys the magic.

 

How about drying fruit. Fan only vs dehydrators vs different temps. Or making tea at what temps. Or how to store your cubes - what makes 'em go bad? Lots of potential for discovering useful information vs subjective recommendations. But if it was easy we would be doing this already.


Edited by raymycoto, 15 January 2019 - 12:38 AM.


#31 PistolPete13

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Posted 15 January 2019 - 01:36 AM

The method so far (I am always looking for improvements);

 

I will have a some sterilized ice cube trays with lids I will write a number on each cell on the tray, then quarter fill with sterile distilled water or even LC.

 

I will also have a drying tray that has little numbered sections.

 

  • I plan to harvest a fresh fruit, tear it open in front of the hood and take a tissue culture, which would be dropped into (for example) cube number 1 on the tray.
  • The remains of that specimen would be placed into the compartment on the drying tray with the same number(to keep track of whats what). And then dry over night.
  • In the morning the bone dry sample would be powdered and 100mg weighed on my analytical scales that weigh down to 1 milligram.
  • The 100mg sample put into a 2ml eppendorf tube
  • The pipettor is then used to transfer exactly 1000ul of methanol into the tube, the pipettor is adjustable down to the microliter!
  • The lid on the tube snap shut, the sample would then be shaken and left for a predetermined amount of time.
  • The tube would then be opened and small wad of cotton wool about the size of the mouth of the tube is put in.
  • The cotton wool is used like a filter/coffee plunger using the capillary tube to push it down and draw exactly 2ul of sample. Microcaps are extremely accurate!
  • The sample spotted on the plate, and then its run.

I have left out a lot of little details but that is basically where I was going.

 

Yes the potential for testing extracts is massive, especially since a lot of the Rf values and compounds that occur in the fruiting body are well known. So it would be easy to see if it was Urea for example that caused the stomach discomfort, dont forget substrate and additive tests!

 

I am not arrogant but I honestly think I have got this, it is not my first time using TLC. And I have heaps of literature that I have been collecting for awhile. If we fail here and document everything it will surely pave the way for the person that gets it anyway.....

 

 



#32 WalkingCatfish

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Posted 15 January 2019 - 07:44 AM

This is potentially important work, PP13! I'm really impressed with your determination and focus.

 

I have good access to literature, so if there are any papers you need, I might be able to help.


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#33 raymycoto

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Posted 15 January 2019 - 09:01 AM

Well, sounds pretty cool. Maybe do a simple run of just a couple of fruits. Perhaps mature vs aborts or just a couple different ones plus a control of some sort. Don't get held up with the 1 part in 1000 precision unless you already have the equipment laying around but just go through the process - dry the fruit, pulverize, MeOH, apply to the plate, run TLC, scan and let us know how it goes. Looking forward to hearing more.


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#34 DaveyJonez

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Posted 15 January 2019 - 09:14 AM

A rehashing the above, but my $0.02

I believe there is 'tolerance' to entheogens, psilocin, dmt, etc, at three levels.

  • Receptor level - The biochemical level. The entheogens as are seratonin receptor agonists, tryptamines, substituted dimethytryptamines, that is. And as such their action at the 5HT2a receptor is quickly down regulated as with many catecholamine receptors.
  • Brain level - their action at the 5HT2a receptor is to provide antagonistic action along other pathways, hence the term "shutting down the default mode network (DMN)". I don't doubt that there is some way that the brain fights back to reestablish these primitive pathways, thus perhaps tolerance may occur or other higher, yet subcognitive levels.
  • Conscious and behavioral level - most of us are all familiar with the factors of set and setting which influence a favorable response to entheogenic treatment. I can't help but think that increasing interval between dosing does not lead to a more meaningful or beneficial setting. We 'need' the therapy more as time goes by but also with time we assimilate what we have learned and implement cognitive and behavioral changes. If we are lucky we have realized some favorable feedback not only in through our own mindful assimilation but perhaps through societal interaction - family or work, for example. Dosing at too short intervals may short cut this important process of assimilation.
Have respect for the power of these truly unique drugs. Wait and savor opportunity to redose. Be grateful that you have both access to them as well as the knowledge and discipline to use them for good. Teach others to do the same.
This is the lesson I've learned from my current, seemingly impenetrable tolerance.. I've decided to wait 3 weeks for my next session.. Do you think this is enough time for full "recalibration"?

#35 raymycoto

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Posted 15 January 2019 - 10:26 AM

Yes, a couple of weeks would be good as well. You might enjoy this book. I think it's even available on pdf somewhere but I really enjoy audio books.

 

https://www.audible....book/B07BRBB739


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#36 DaveyJonez

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Posted 15 January 2019 - 11:50 AM

Yes, a couple of weeks would be good as well. You might enjoy this book. I think it's even available on pdf somewhere but I really enjoy audio books.

https://www.audible....book/B07BRBB739

Thanks for the recommendation..im definitely going to read that.. I'm very glad it'll only be a couple weeks as spending time with Cubensis is absolutely my favorite weekend activity.. I guess I have one more question concerning tolerance; once I shake off my tolerance, if I only take mushrooms once a week will I still accumulate tolerance or do I always have to wait 2 weeks to get back to baseline?

#37 raymycoto

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Posted 15 January 2019 - 12:37 PM

Would be hard to say. Depends on the dosage and such as above. Some would say that a full dose more than a few times per year is a bit much for a meaningful experience. But perhaps that book or others' opinion would help. Certainly would vary from one individual to another. Fadiman does address that.

 

Another great book is Michael Pollan "Change your Mind" re psychedelics

And while you are at it, entirely different but everyone should read - Michael Pollan - Omnivore's Dilemma - outstanding unbiased personal research into the history, politics, economic, science and ethics of our current nutritional dilemma.


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#38 Deleena24

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Posted 15 January 2019 - 01:05 PM

Yes, a couple of weeks would be good as well. You might enjoy this book. I think it's even available on pdf somewhere but I really enjoy audio books.

https://www.audible....book/B07BRBB739

Thanks for the recommendation..im definitely going to read that.. I'm very glad it'll only be a couple weeks as spending time with Cubensis is absolutely my favorite weekend activity.. I guess I have one more question concerning tolerance; once I shake off my tolerance, if I only take mushrooms once a week will I still accumulate tolerance or do I always have to wait 2 weeks to get back to baseline?

I can say with confidence that if you're only taking mushrooms, no other hallucinogenic substances to get cross tolerance, you can dose once a week and get the full experience.
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#39 DaveyJonez

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Posted 15 January 2019 - 01:37 PM

Yes, a couple of weeks would be good as well. You might enjoy this book. I think it's even available on pdf somewhere but I really enjoy audio books.

https://www.audible....book/B07BRBB739

Thanks for the recommendation..im definitely going to read that.. I'm very glad it'll only be a couple weeks as spending time with Cubensis is absolutely my favorite weekend activity.. I guess I have one more question concerning tolerance; once I shake off my tolerance, if I only take mushrooms once a week will I still accumulate tolerance or do I always have to wait 2 weeks to get back to baseline?
I can say with confidence that if you're only taking mushrooms, no other hallucinogenic substances to get cross tolerance, you can dose once a week and get the full experience.
That's what I like to hear!

#40 DaveyJonez

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Posted 15 January 2019 - 01:40 PM

Would be hard to say. Depends on the dosage and such as above. Some would say that a full dose more than a few times per year is a bit much for a meaningful experience. But perhaps that book or others' opinion would help. Certainly would vary from one individual to another. Fadiman does address that.

Another great book is Michael Pollan "Change your Mind" re psychedelics
And while you are at it, entirely different but everyone should read - Michael Pollan - Omnivore's Dilemma - outstanding unbiased personal research into the history, politics, economic, science and ethics of our current nutritional dilemma.

I actually have "how to change your mind" but I haven't gotten very far in it yet.. I'd have to disagree with the notion that more than a few times a year is too much for a meaningful experience, I personally believe that as long as there is respect and openness to the experience there really isn't a spiritual limit.
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