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OfflineNoOneKnowsHowToAct
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Re: Potency Project [Re: Necco]
    #11791458 - 01/08/10 05:55 PM (15 years, 11 days ago)

I want to do this to breed potency. How? Let me explain (assuming we can determine potency through extraction and weighing)

1. I get my first flush from a MS grow and take let's say 4 ideal shrooms from the batch and clone them.

or

1. I use Agar to isolate let's say 4 different isolates from a MS syringe.

2. I grow out the isolates each in let's say 3 separate trays (ideally) (we would have 12 trays total) all under the same conditions as well.

3. I weigh the first flush of each tray.

4. I take a sporeprint from each isolate.

5. I combine the flushes from the same isolate and perform an extraction. 4 extractions total, One for each isolate.

6. I weight the amount of residue or crystals from each extraction.

7. I divide the amount of extract by the amount the total weight of that isolate's flush (the starting matter for the extraction) to get the percentage of psilocybin in the matter.

8. I use the sporeprint for the isolate with the highest percentage to start over at #1 as my MS source.

I then repeat this process a hundred times. This is simple selective breeding and it occurs in nature. It's why cubes from Florida look and grow differently than ones in Thailand for example...

I don't see why it wouldn't be possible to breed for potency like this and eventually after enough runs obtain something that is as genetically unique as say PE.


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OfflineRoseM
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Re: Potency Project [Re: NoOneKnowsHowToAct] * 1
    #11797049 - 01/09/10 03:48 PM (15 years, 10 days ago)

SO we need some volunteers.

Who will make extractions from 5-10% of their grow?

We need to agree upon an extraction technique (a simple extraction technique) so results can be somewhat regulated.

I am willing to compile all the information, as long as it is sent to me.

How do we do this?


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Offlineevildee125
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Re: Potency Project [Re: Rose]
    #11797118 - 01/09/10 04:02 PM (15 years, 10 days ago)

i have b+ but not pe.. i could compare to the other cubes i have.. that about it..that should atleast say something, however small, about them being bunk, do you have a suggestion to a good extraction tek


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Re: Potency Project [Re: evildee125]
    #11797135 - 01/09/10 04:04 PM (15 years, 10 days ago)

Well, honestly I think just eating them is enough to give a yes/no answer. We aren't doing drug approval trials- we already approve of this stuff. :thumbup:
If you find a batch you think are bunk, try them on a friend and see what they think.


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OfflineLogicaL ChaosM
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Re: Potency Project [Re: evildee125]
    #11797143 - 01/09/10 04:06 PM (15 years, 10 days ago)

I would say B+ is a good "control" mushroom species to compare PE to.

I've never grown B+ (I grow Mexican-type species), but from what I've heard, B+ is well-known and a "classic", and average strain. I'd say do the extraction on your B+, if you are cool with sacificing your yields for science. Make sure you take a random sample of the mushrooms you pick to keep the results valid. How can you randomly pick mushrooms, would putting them in a flat dish/pan that's over your head (someone holds it high), then reaching in and picking one, work?

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Offlineevildee125
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Re: Potency Project [Re: Doc_T]
    #11797156 - 01/09/10 04:09 PM (15 years, 10 days ago)

Quote:

Doc_T said:
Well, honestly I think just eating them is enough to give a yes/no answer. We aren't doing drug approval trials- we already approve of this stuff. :thumbup:
If you find a batch you think are bunk, try them on a friend and see what they think.



i like this idea.. not to mention .. i have a fair number of guinea pigs:smile:


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OfflineRogerRabbitM
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Re: Potency Project [Re: LogicaL Chaos]
    #11797172 - 01/09/10 04:12 PM (15 years, 10 days ago)

Quote:

LogicaL Chaos said:
How can you randomly pick mushrooms, would putting them in a flat dish/pan that's over your head (someone holds it high), then reaching in and picking one, work?





Grind the entire flush to a powder and then take the desired amount.  This averages out the differences between fruits.
RR


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OfflineLogicaL ChaosM
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Re: Potency Project [Re: RogerRabbit]
    #11797185 - 01/09/10 04:15 PM (15 years, 10 days ago)

Oh yeah, I think I had that same idea in an earilier post I had about weighing the powder on a scale before extracting.

What about between 2 different flushes: is the first flush more "potent" the second or thrid? Are the active levels pretty consistant between sequential flushes?

:strokebeard:
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Re: Potency Project [Re: evildee125]
    #11797192 - 01/09/10 04:16 PM (15 years, 10 days ago)

Quote:

evildee125 said:
Quote:

Doc_T said:
Well, honestly I think just eating them is enough to give a yes/no answer. We aren't doing drug approval trials- we already approve of this stuff. :thumbup:
If you find a batch you think are bunk, try them on a friend and see what they think.



i like this idea.. not to mention .. i have a fair number of guinea pigs:)




Right. Maybe we'll get a big fat "no difference" from the first 100 or so people, and that is the end of that.
Of maybe we'll get a fair number that notice a difference, and it correlates well with what we expect.
Then we go on from there with a more refined method.
But we don't need chemically pure dose in lab conditions to answer the basic question.


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OfflineRoseM
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Re: Potency Project [Re: LogicaL Chaos]
    #11797199 - 01/09/10 04:17 PM (15 years, 10 days ago)

Grinding is God!


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Re: Potency Project [Re: Rose]
    #11797896 - 01/09/10 06:26 PM (15 years, 10 days ago)

Grinding is fine to average out a batch of dried mushrooms, but we have to make sure we have a standardized drying technique as well. Then we have to make sure that we all have mushrooms with the same residual moisture; even when they seem dry they have plenty of bound water which could vary from person to person and affect the results.

I'm sure there are plenty of people around here willing to off a portion to test with, including me. The hard part will be finding people willing to set out pans of gasoline evaporating in their kitchen.

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Offlinefundamentalchair
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Re: Potency Project [Re: Necco]
    #11798030 - 01/09/10 07:02 PM (15 years, 10 days ago)

Feel free to TL;DR this mindless babble.

I have quoted the following people in this post:
mubba
Cervantes
LogicaL Chaos
ImaginingEmotions
evildee125
Doc_T
SillyBilly
RR
NoOneKnowsHowToAct

Quote:

mubba said:
Using fresh material would be essential due to differences in drying technique and the affect of drying on actives.




If done at the same time, drying conditions would be identical (assuming done in one lab, not as a collective project).

I'm concerned with water content ratios, as I think it would be most benificial to express the psilocybin/psilocin content per mycelia mass not per mycelia mass plus unknown water content.

Quote:

Conversion of psilocybin to psilocin would be unnecessary using colorimetric techniques as I described earlier.




I am not familiar with those processes or their abilities and/or limitations. Are they able to differentiate between the two substances and be able to give some way of inferring the mass of each?

Quote:

Harvesting neon is preposterous. You could accomplish the same goal by using CO2 from a draft system, you could even use natural gas from your stove. Inert gasses are unnecessary in this case, only low reactive gasses.




I had only suggested a noble gas due to it's relative possibility to be obtained in a clandestine and fairly inexpensive manner. Also, I would think it to be prudent to stay away from hydrocarbons. CO2 may be a good alternative, although I question the purity of the cartridges used for 'mini-kegs'.

Quote:

Lol, anhydrous sulfuric acid. Try mixing dilute sulfuric acid with dilute sodium hydroxide, drying it out in a pan then baking it in the oven. Tada!




Wouldn't that form a salt?

Quote:

Cervantes said:
I am not convinced substrate effects potency, but it would be worth researching... even if it just puts that rumor to rest.




Until one could prove if the contents of the substrate have an effect on potency, it would be difficult to draw valid conclusions without using identical substrates between grows. This could be mitigated though the use of a large sample size, but could well cause less than ideal data.

Substrate has to have some effect on potency, although I'm not sure of any concrete evidence to outline possible factors. Simply, a phosphorus devoid substrate would have a hard time producing Psilocybin. By having multiple concurrent with identical (as possible) substrates/fruiting conditions/etc. would give data that would be least open to interpretation due to uncontrolled variables.

Quote:

LogicaL Chaos said:
in those indentical conditions.




Impossible to do between grow to grow. This must be done at the same time to ensure the best possible identical conditions.

Quote:

similiar looking fruits, the ones that look like how they are "supposed" to look like.




Statically speaking, that's not a 'random sample'. It would be more accurate to test the entire population. Since 'supposed to look like' is entirely subjective, especially without valid metrics, it wouldn't necessarily be a good selection criteria.

Quote:

...wet weight, trim stems to get the same weight.




Wet weight is a poor indication of active ratio to mycelia. Dry mass to wet mass ratios can be different between races and between individual samples of the same race. This leaves an untraceable variable of water content per fruit.

Doing calculations as active content per mass would make the 'identical weight' think unnecessary. Furthermore, if there is any difference in potency per mass of stem to cap, you are artificially skewing your data by trimming the stem.

Quote:

Since the extraction is a collection of mushrooms from the same "strain", it should be a pretty good average, at least for that flush.




Unknown and possibly invalid assumption. It would be more accurate to just grab all the fruits and find their active/mass ratio and use that as your 'average'.

Quote:

After the extraction, dry the psilocybin crystals completely, then weight them on a precise scale (0.001 grams or more resolution).

If the amounts are the same, do a different extraction process (using the same growing techniques) of the next flush that extracts just psilocin.




I am not aware if it is possible to separate psilocybin from from psilocin. Even so, psilocybin may decay into psilocin at an unknown rate possibly throwing off your calculations.

If it is possible to separate the 2, it would be best to measure both from the same 'sample'.

Quote:

And if those 2 values are the same amount, then you can do a subjective potency test.




Are you saying that if the psilocybin and psilocin ratio are close between extractions of a singular active between 2 separate flushes would then allow for a subjective test? I don't follow.

Quote:

First, weight 2 dry fruits from 2 different strain, PE included, and cut the stem to equal the amounts. Make sure they "look" like classic forms of their respective strains. Then, tell 2 people they are recieving reg. mushrooms with reg. potency by showing them both a regular looking shroom, then prepare a method of eating it so that they won't know how much or what mushroom they got, such as a fruit smoothie.




Make sure it's double blind. The 'giver' of the 'smoothie' would have to not know which is which, and all personnel would have to be not in contact with the person who does through the experiment.

It would be helpful to have some sort of metric for someone to be able to describe their experience. Since most people will fall back to explaining things from their past experiences and not everyone's experiences are identical it would make for a difficult comparison between 2 people.

You could do a modified form of the experiment that is done in DMT: The Spirit Molecule by doing multiple repetitions of differing doses, each taking each mushroom individually as well as a placebo or two. Oh yeah, don't forget the double blind thing.

Quote:

What do you think, would this work?




With some modification, possibly.

Quote:

ImaginingEmotions said:
If people can contest that they have had extreme differences in potency, it would be well known to begin with and im pretty sure people would be cultivating that strain more than any other.




Define 'well known'. Until someone has some data through experimentation, all off this 'knowledge' is circumstantial at best. That is the entire purpose of this discussion.

All other things being equal, yes, it would most likely be the most popular. There are other considerations such as overall output weight, ease of growth, colonization speed, et cetra ad nausem that would make that statement not true.


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Edited by fundamentalchair (01/09/10 07:05 PM)

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Offlinefundamentalchair
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Re: Potency Project [Re: fundamentalchair]
    #11798043 - 01/09/10 07:04 PM (15 years, 10 days ago)

Quote:

evildee125 said:
yes ive been here long enough to see you and others say this many times.. i was thinking more along the lines of the importance of additives, for instance, that reportedly balance lack of nutrients.. and how levels of nitrogen correlate to potency.. from what ive seen.. the additives dont really seem all that necessary..




I would take a walk out on a limb and say that in any substrate capable of supporting shroom life most likely has enough nutrients to exceed the biological efficiency of active production. Even in the case that it doesn't, not many people have the testing equipment to measure that sort of minute changes between substrates. Strain isolation would most likely give much higher results than adding random crap to your substrate given unknown forms of reagents and catalysts that would be beneficial. To 'figure' this out would be a lot of work, much more than testing relative potency.

Quote:

Doc_T said:
And before people start in with "controls, precision, etc"- I know what a gram of shrooms feels like, guys.




I wouldn't contend that you don't. I would contend that there are a great deal of variables that wouldn't be excluded through such methodology. The difference in baeocystin alone could cause a major difference.

At any rate, dosing up some PE and stating that you feel it is more potent than another cube wouldn't get us any farther than we are right now. I don't mean it to seem that it isn't useful, but it isn't an actual measurement.

Quote:

SillyBilly said:
Could also be a super conspiracy by some smart drug dealer who realized the density of PE usually resulted in better weights for yield so he created a giant placebo effect scam to trick buyers into wanting PE only. Making him increase profits.:awebig:




I've heard people say that the gold flecks are an indication that the shrooms are potent. Back in my drug purchasing days, I've never ran across PE nor heard any 'stories' about it.

This 'potent PE' seems to be more spread by growers than anything; at least when I bought shrooms someone could have sold me a bag of dried shataki and I wouldn't have known the difference (pre-consumption).

Quote:

evildee125 said:
so you would be testing the best genetics one was able to obtain from one strain vs the best genetics you can obtain from another... youd still have to create several isolates from several sources wouldnt you..
youd still be looking at averages.. isolates dont make sense for what the project is trying to achieve




They very well would, if done correctly. Multiple experiments would have to be done over various prints from different sources to lend conclusiveness, or better done concurrently.

Assuming unlimited resources (or at least a lab, a team, some serious equipment, and lots of room), I could see the benifit to fruiting out all isolates from several prints of each selected 'race' as well as comparing it to several MS of each.

Quote:

Cervantes said:
RR's not bullying anyone. He's participating in this discussion in the same cranky way he participates in all discussions. He knows his shit and if I were a betting man, I'd put my money on his theories.




Whats the emoticon for 'you hit the nail on the head'?

Quote:

That said, I agree, the point is to compare PE to the average cube... and that means we've also gotta' figure out what an 'Average' cube is.




Median would be a much better number to work from, rather than mean.

Quote:

MS, and not isolates are what we need in order to find this info.




You have the right idea but the wrong process. Multiple MS inoculations may not give a median or average results as compared to the median of the contents of all isolations. In short, with MS you don't know what the hell your gonna get or if it's even an accurate representation of 'average'. By compiling data from all isolates from a spore sample as well as multiple MS shots from the same sample you can then determine if that is an accurate correlation.

My guess is that the confidence level required to use MS as 'average' would be quite low. This of course could be mitigated by many iterations of MS grows, but it sounds tedious and less precise than just doing the isolate route.

Quote:

RogerRabbit said:
One of these days, somebody is going to actually do it instead of just talking. :rolleyes:
RR




Hey man, if I were independently wealthy and had an extra 50 grand and a year to blow, I'd be on it like chronic.

Quote:

Cervantes said:
I understand how isolates would help in a true, scientific environment. This unfortunately is not that environment. It is The Shroomery. If we opened this experiment up to everybody's isolates, we'd get uneven potency readings because many cultivators isolate FOR potency.




If that were true, I don't think B  would have a rep of being less potent. Who does isolates and has testing equipment to verify the potency of each isolate without taking a tremendous amount of time (i.e. not just shrooming twice a week till all 20-30 isolates have been tested)?

The only logical way to do such is to work with all the isolates.

If you're worried about non-conformity between isolation parameters, how do you address the conformity issues between the other variables such as extraction technique, substrate variance, and the homogeneity of ingredients used by different people?

Quote:

If we opened it up to MS cube grows and MS PE grows, we may see something a little more 'Common'... especially if we have many people participate.




While multiple iterations done by multiple people can well be valid, there are a great deal of things that need to be considered when such an activity is undertaken. In the scientific community extensive documentation and procedure is followed to be able to replicate the same circumstances as best as possible. Using different suppliers for ingredients with unknown or unverifiable purity or contaminants and the possibility of inaccurate measurement devices makes this a very difficult undertaking to get any conclusive data from.

In the interest of academic honesty, these issues will have to be addressed to at least understand the possible variance between testing done from individual to individual. It would be rather shameful to present inaccurate evidence as accurate without stating its probable inaccuracy.

Quote:

mubba said:
This is a classic case of the vaguery and detachedness of forum posting leading nobody to understand what anybody else is trying to get at. In other words, plenty of people are reading into people's posts the wrong way and/or totally ignoring them.




I believe this is a problem due to people would rather be misunderstood than ignored. TL;DR tends to pop up quite a bit if someone is verbose enough to describe their thoughts concisely.

Quote:

This thread has branched out so much, it is frustrating to read. So many nay-sayers and people with different ideas who are not willing to work together provide that we are sure to make no progress as a community. I think we can only wait until one dedicated person or small group does all the work on their own and wows us with the results. Bummer, I guess this makes me a nay-sayer.




This is the reason that instead of world peace we bomb things. It's just easier that way.

Quote:

TLC doesn't require anything fancy, it requires a couple plates of glass, some silica gel and some plaster of paris. A mass spectrometer wouldn't be necessary, just the chromatograph attached. Even GC isn't that great because of the heat.




Mind elaborating on this process or at least posting a link so that I can better understand this process and it's abilities/limitations?

Quote:

mubba said:
I agree that if anybody wants consistent results we need an isolate, but this thread isn't about maintaining a consistent culture, it is about the average expectation between lines.




Imagining a MS as the proverbial dice roll, you will never roll a '3.5'. For a dice the only sure fire way to figure out the 'average' roll would be to calculate the 'value' of each outcome and multiply it by it's probability. The sum of these values would then be it's average.

While I am in full understanding that I am vastly oversimplifying by making a comparison from a MS shoot to a dice roll, the same basic premises apply. One 'roll' won't be average, and without doing some sort of statistical analysis on multiple samples (as finding every possibility would be impossible), your comparison would be worth it's weight in ice; To an Eskimo.

By measuring and compiling the data from all isolates obtained from a print (or multiples) I think it would give better data as well as better measurements and plots of plausible outcomes. I honestly can't see the with this process other than the large scale of work that would need to be done.

Quote:

NoOneKnowsHowToAct said:
I don't see why it wouldn't be possible to breed for potency like this and eventually after enough runs obtain something that is as genetically unique as say PE.




Well, I mean, that's a rough overview of genetic engineering 101. If the rumors of the origins of PE are remotely true, I'm quite sure that's exactly what happened over the course of 10-15 years.

How does one quantify 'unique'? I mean, I'm unique just like everyone else.


--------------------
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InvisibleNecco
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Re: Potency Project [Re: fundamentalchair]
    #11798135 - 01/09/10 07:23 PM (15 years, 10 days ago)

Quote:

fundamentalchair said:
Quote:

mubba said:
Quote:

Conversion of psilocybin to psilocin would be unnecessary using colorimetric techniques as I described earlier.




I am not familiar with those processes or their abilities and/or limitations. Are they able to differentiate between the two substances and be able to give some way of inferring the mass of each?






I believe colorants are available which are selective for indole. In this case, using spectrophotometry would give us an indication of the total of all indole containing molecules in the extract, regardless of side groups etc. We would not be able to use this to figure out how much of each were there, just the total indole centers. I'd say that would be pretty useful info, since it would be independent of molecular mass.

Quote:

fundamentalchair said:
Quote:

Harvesting neon is preposterous. You could accomplish the same goal by using CO2 from a draft system, you could even use natural gas from your stove. Inert gasses are unnecessary in this case, only low reactive gasses.




I had only suggested a noble gas due to it's relative possibility to be obtained in a clandestine and fairly inexpensive manner. Also, I would think it to be prudent to stay away from hydrocarbons. CO2 may be a good alternative, although I question the purity of the cartridges used for 'mini-kegs'.



I was referring to the CO2 you get in a tank for pressurizing kegs in a draft system, like at a bar (or my garage). It is very pure, and has to be, given that it is for human intake the purity standards are very strict. I would still prefer something other than CO2 because of its acidity and capability to form salts, and I would prefer something other than hydrocarbons (natural gas) only because they can catch on fire. I was just throwing stuff out there, but really it would be easy to rent a N2 tank, except a lot of folks here don't seem to even have $25 to spare.

Quote:

fundamentalchair said:
Quote:

Lol, anhydrous sulfuric acid. Try mixing dilute sulfuric acid with dilute sodium hydroxide, drying it out in a pan then baking it in the oven. Tada!




Wouldn't that form a salt?



I was responding to somebody else's suggestion that in order to make sodium sulfate you would use anhydrous sulfuric acid (LOL). Yes, the method I posted forms a salt, but that was the end goal (for drying the non polar extract).


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Re: Potency Project [Re: Necco]
    #11798238 - 01/09/10 07:43 PM (15 years, 10 days ago)

Quote:

Cervantes said:
SO we need some volunteers.

Who will make extractions from 5-10% of their grow?

We need to agree upon an extraction technique (a simple extraction technique) so results can be somewhat regulated.

I am willing to compile all the information, as long as it is sent to me.




Tell me what and how to do it, and I can get you back some data. Won't be from B+ or PE, but it'll be from a cube.

Quote:

LogicaL Chaos said:
How can you randomly pick mushrooms, would putting them in a flat dish/pan that's over your head (someone holds it high), then reaching in and picking one, work?

~ LogicaL Chaos ~




Personally, I'd just stick a bunch in a blender and take out some and use the rest for chocolates. Assuming that is a valid way for the scope of this experiment.

Quote:

Doc_T said:
Right. Maybe we'll get a big fat "no difference" from the first 100 or so people, and that is the end of that.
Of maybe we'll get a fair number that notice a difference, and it correlates well with what we expect.
Then we go on from there with a more refined method.
But we don't need chemically pure dose in lab conditions to answer the basic question.




Variables man, variables. Historically speaking non blind experiments tend to produce data that supports the hypothesis of the experimenter, either intentionally or unintentionally.

I got $5 on it being a complete crap-shoot with too many outliers to get anything resembling a consistent data-set.

Quote:

mubba said:
I'm sure there are plenty of people around here willing to off a portion to test with, including me. The hard part will be finding people willing to set out pans of gasoline evaporating in their kitchen.




not like it'd be the first time...

Quote:

I believe colorants are available which are selective for indole. In this case, using spectrophotometry would give us an indication of the total of all indole containing molecules in the extract, regardless of side groups etc. We would not be able to use this to figure out how much of each were there, just the total indole centers. I'd say that would be pretty useful info, since it would be independent of molecular mass.




So you're doing a count of actual molecules, or an approximation of such? Assuming it isn't expensive, I surely wouldn't be opposed to a mass-less measurement of actives. I guess the only question would be how many/much inactive indoles are present in shrooms and do/can they vary from race to race in non-linear correlations from actives.

Quote:

I was referring to the CO2 you get in a tank for pressurizing kegs in a draft system, like at a bar (or my garage). It is very pure, and has to be, given that it is for human intake the purity standards are very strict. I would still prefer something other than CO2 because of its acidity and capability to form salts, and I would prefer something other than hydrocarbons (natural gas) only because they can catch on fire. I was just throwing stuff out there, but really it would be easy to rent a N2 tank, except a lot of folks here don't seem to even have $25 to spare.




What I was referring to is pretty much the same thing but designed for like 1 gallon kegs. They're like really big whippets (16g cartridges). I huffed one :facepalm: and got WAY out of breath (as I expected to) but I got this really weird burning of the throat from it that I totally didn't expect.

Quote:

I was responding to somebody else's suggestion that in order to make sodium sulfate you would use anhydrous sulfuric acid (LOL). Yes, the method I posted forms a salt, but that was the end goal (for drying the non polar extract).




For some reason I thought you were trying to make anhydrous sulfuric acid from dilute and lye.

I mentioned some anhydrous acid earlier, although I meant it in terms of converting psilocybin to psilocin. That's probably completly not the context that I typed it in; probably one of those times my stupid fingers get confused by the ramblings of my confused brain.

Sadly I'm smart enough to know that I'm too stupid and inexperienced to still out a acid, but I'd do it anyhow. Luckily enough, I don't own much in the way of glassware.


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Re: Potency Project [Re: fundamentalchair]
    #11798328 - 01/09/10 08:01 PM (15 years, 10 days ago)

Wow, great analysis of our posts chairman :wink:

OK, to re-cap, here's the procedures we need to do (Correct any incongurancies found)

From the Beginning with Spores

(1) Spores Source: Should be from PE and a Control strain (whatever we agree on as a good Control strain, as a group), that have both been isolated from their respective strains. Spawn run/substrate has to be MS inoculation from those isolates.

(2) Substrate: Have to be identical as possible, like water, nutritious material, moisture material, etc. Also, the colonization conditions, light-exposure, fruiting chamber, and sterile techniques should be standardized so they are identical as possible. Also, the PE and Control isolates have to be grown simultaneously, so that ambient weather of the grow room does not affect the results.

(3) Sampling Fruits: Take ALL the mushrooms from just ONE flush of each of the 2 strains and dry them all under identical drying conditions. Use accurate hydrometer/temp. gauges to measure drying conditions. Drying of each strain must be done together.
    After drying, grind up to a fine powder, controlling for loss of powder and error caused by losing material to the grinder and the air (tends to float). ***Somehow, the material from each mushroom has to be thoroughly mixed, so that its evenly distributed inside the total powder collection.*** Maybe a tumbler of some sort to evenly-mix all the powder, so if a sample is taken from the total, it represents every mushroom from the flush (at least in theory).
Also, what should be the level of Error be? Hopefully its not so high its the difference between the concentrations of actives between PE and the Control...That would be troublesome.

Questions:
Does potency vary "enough" from 1 flush to next to consider taking an average of multiple flushes of each strain? Should we just use the first flush for simplistic sake?

(4) Weighing: Just the Powder
Using a precise (0.01g resolution or better), accurate scale, under the same ambient conditions to control for moisture affecting the weigh of the "dried" samples. There could be 2 different methods to do this, Simple and Advanced.
The Simple method would consist of taking all the dried mushrooms from 1 flush and powdering them in a blender, then weighing the TOTAL amount of dry yield collected from that strain and flush.
The Advanced method would be to weigh each dried mushroom collected from the flush independent of each other, than record those values. After that, those individual mushroom weights would be total up and then used in the "Statistical Analysis" phase, which is really just using math with data sets.

(5) (***For Advanced Study***) Statistical Analysis of the mushroom's dry weight.
For everyone who was performing the Advanced Method of weighing discussed above, all their data values for each mushroom who be organized from lightest to heaviest, then the Median value would be found. This would give a pretty good value of the "average" weight of a mushroom. Not useful for determining potency directly, but it could provide interesting results (ex: PE fruits are heavier than B+ fruits, on average).

(6A) (***For Advanced Study***) Extraction of Psychoactive Chemicals from Powdered Yield
First of all, the extraction(s) of the psychoactive chemicals should be simple, cheap, easy-to-find materials. Since no solvent can extract all psychoactive chemicals simultaneously, multiple extractions using different chemicals must be done. Then, once the simplest and cheapest extractions are determined, rules must be define to standardize it so it gives good data results.
- After each psychoactive chemical is extracted, each of the 3 chemicals should be completed dried (in the same humidity levels for everyone in this study) and then weighed on a precise, accurate scaled (like discussed above).
Then, the weight of each chemical should be recorded and then add to a community-wide data set explained below... 


(6B)
(***For Advanced Method***) Statistical Analysis of the extracted psychoactive components from powdered flush (the total mixture)

From everyone who participates in the "extraction of active chemicals" part of the experiment, we could gather everyone's "Total Weight of Active chemicals" for their respective strain (PE or the Control), and then take the Median of those data values, and use that as our "average weight of active chemicals in X strain". This statistical analysis will have to be done for each of the 3 psychoactive chemicals: Psilocybin, Psilocyin, and Baaocin. The specific chemical extracted (since all 3 cannot be extracted with one solvent) have to be put into separate data sets. Once the data sets are made for each of the 3 chemicals mentioned, the median of each chemical's data set would be taken, which would represent the "average amount" for the given strain.

-> Assuming extraction technique is explicitly explained, standardized, and easy to perform.

(7) Last Part of Project: The Subjective Trip Experiment: Has to be a double-blind, well-regulated psychology experiment. First, the testee's cannot know what the experiment is about, which is the potency of cubenis strains. Second, they cannot know which strain they are getting (or even that PE is a possible strain they could receive). Third, they cannot know how much they are receiving (to be ethical, no heroic doses should be given!) or what it looks like (also a factor and the precipitants giving the doses must NOT know which strain they got. Better yet, they shouldn't even know they are getting dosed with mushrooms! But, they have to be told its a hallucinogen or that would be unethical. That should really control for some subjective variables, but it wouldn't reflect "naturalistic settings" of the causal shroomer.

-> Then a standardize "Trip Report" had to be made so that the experiencer can describe his/her experience into a "quantitative" method of measuring it. How to quantify the psychological effects of a psychedelic on a person, which can be standardized to all people, is going to be tricky, to say the least. 

My Comment: I think this would be the hardest to get "good" data from because there are so many variables on top of the variables in the above extraction process. Of course, you could skip the "extraction of psychoactive components" process I described in (6) and go straight to dosing people with the weighed powder. But once you go past the numbers, like weighing extracted active chemicals, but its going to get subjective. But, in reality, that's really the only way to measure "potency", because potency, by nature, has to do with the psychoactive intensity of chemicals relative to how much mushroom material you actual ingested. But Remember, this is a psychedelic, and by nature, psychedelics are unpredictable. :mushroom2:

I think that's it, did I miss anything?

Whew!
~ LogicaL Chaos ~

Edited by LogicaL Chaos (01/10/10 09:06 PM)

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Re: Potency Project [Re: fundamentalchair]
    #11798334 - 01/09/10 08:01 PM (15 years, 10 days ago)

Quote:

Quote:

TLC doesn't require anything fancy, it requires a couple plates of glass, some silica gel and some plaster of paris. A mass spectrometer wouldn't be necessary, just the chromatograph attached. Even GC isn't that great because of the heat.




Mind elaborating on this process or at least posting a link so that I can better understand this process and it's abilities/limitations?



Which method would you like me to get into, I listed three? None of them, if you ask me, are appropriate for our purposes (especially considering that we can't use each other's equipment).

TLC is really easy to set up, and "easy" to perform, but at the same time is a poor method for macroscopic separations and it can be very difficult to duplicate results. I can supply links to plate preparation, if you want, but I don't think we should bother with this method.

As for the other techniques: Mass Spectrometry is for identification not quantification, so it isn't necessary. GC aka GLC (in most instances) is performed by injecting a sample into a heated port where the sample is vaporized then pushed through a column by a heated inert gas. This is Kosher if you are using MassSpec and you can see the fragments, but I'd prefer to analyze my molecules in one piece. Few of us have access to this equipment, and for those of us that do, I'd walk the next room over to the HPLC/UVvis <sigh, if only>

I'm all for spectrophotometry. Somebody send me a spectronic and samples and I'll get to work. I'm sure one of you undergrads has access to your school's surplus/outdated specs :wink:

Quote:

fundamentalchair said: (in reference to colorimetry)
So you're doing a count of actual molecules, or an approximation of such? Assuming it isn't expensive, I surely wouldn't be opposed to a mass-less measurement of actives. I guess the only question would be how many/much inactive indoles are present in shrooms and do/can they vary from race to race in non-linear correlations from actives.



Yes, a count of molecules (concentration in solution). Approximation, certainly, but hopefully close. I won't pretend to know how many interfering chemicals would be in whatever extract we created, but I'm hoping most would be insoluble (like many proteins) or in low quantities compared to actives. A selective extraction would be necessary if there were a lot of interfering molecules in a simple extract.
The machine used to measure absorbance, a spectrophotometer, is not expensive compared to other analytical machines, but it still costs several hundred dollars (to thousands) new.

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Re: Potency Project [Re: LogicaL Chaos]
    #11798344 - 01/09/10 08:04 PM (15 years, 10 days ago)

Quote:

Questions: Does potency vary "enough" from 1 flush to next to consider taking an average of multiple flushes of each strain? Should we just use the first flush for simplisitic sake?




Whole 'other question. Let's just say first flush.

Quote:

Thrid, they cannot know how much they are recieving



Disagree, I think people should instead be given an amount they choose as normal for them- it varies from person to person. (I'm a lightweight, 2g is plenty for me.)


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Re: Potency Project [Re: Doc_T]
    #11798595 - 01/09/10 08:48 PM (15 years, 10 days ago)

Quote:

(5) Statistics: Take Median of the weighed dried samples from the PE and Control strain, for most accurate representation of each strains population.




This would be great if we were looking to do statistical analysis of fruit production per substrate material. While it wouldn't be poor practice to record the weights of all picked 'shrooms', finding the median weight of them here doesn't do diddly squat.

Reading further into your post, you may be getting at something that isn't explained in a way which I understand.

Quote:

(6) Extraction of Psychoactive Components: This part I'm confused about. Questions: Which solvent(s) would be best for extracting Psilocybin, Psilocin, and Baeocyin with the least amount of loss from the original powder source? Would it have to be multiple extracts using different solvents? What about drying the extract crystals? How do we control for solvents being left in the final product so that they don't affect the weighing results?




At this time, unanswered in any conclusive manner.

Assuming all water is removed from the material, using identical solvent to material ratio between samples in which the solvent has a greater capacity to hold the actives than needed should grab it all in one go. At current, I am not aware of any singular solvent that will remove only the actives and nothing else, leaving a wash of the original solvent with some other solvent that the actives aren't soluble in. If such creature exists, I sure as hell don't know what it is.

The easiest way to control for solvent being left in the final material would probably be to remove it. Any solvent evaporated that leaves a residue should probably not be used when possible. If it can not be avoided, it will have to be thoroughly washed out or have a predictable mass so that it can be accounted for.

Quote:

-> Assuming extraction technique is explicitly explained and standardized: Use same statistic analysis as above: weigh all extracted samples of each strain and arranged in order of least to greatest amount. Then, take median of each data set for its corrisponding strain (PE and Control) and use that as our "average".




Um, what? I'm really getting lost as to what you are accomplishing or trying to accomplish.

Quote:

First, the testee's must cannot know what the experiment is about, which is potency of strains.




That is a morally questionable statement, depending on the context given to the experimenters. I don't see telling them the nature of the experiment as being detrimental to the results.

Quote:

Second, they cannot know which strain they are getting (or even that PE is a possible strain they could recieve).




Morally speaking, they should be told which strains (including a placebo) they have the possibility of receiving and the dosages of such. I see no benefit to withholding that information.


Quote:

Thrid, they cannot know how much they are recieving (to be ethical, no heroic doses should be given!) or what it looks like (also a factor and the parcipitants giving the doses must NOT know which strain they got.




They should not know what the size of that particular does is. There is no reason for them to not know the dosage range.

Quote:

Better yet, they shouldn't even know they are getting dosed with mushrooms!




That is a horribly despicable and morally bankrupt statement. Do not under any circumstances give any person a substance without their express permission. I don't mean to sound like an asshole, but dosing someone unknowingly or with an unknown substance is not only irresponsible and reckless, but downright evil. Seriously.

Quote:

That should really control for some subjective variables, but it wouldn't reflect "naturalistic settings" of the causal shroomer.




If I had to take a stab at it, I would guess that there are as many 'naturalistic settings' as there are shroomers. Such psychedelic drug experimentation is done in a lab environment to remove the variable of setting. Sure, it probably isn't the best to go shroomin in a white walled room with someone staring at you through one way glass, but no one twisted your arm to volunteer either.

Quote:

Disagree, I think people should instead be given an amount they choose as normal for them- it varies from person to person. (I'm a lightweight, 2g is plenty for me.)




Ideally, this experiment would go over multiple dosages and multiple strains. Each person should be given a placebo at least once, a big dose, a small dose, and a few moderate (although varying) doses.


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Re: Potency Project [Re: fundamentalchair]
    #11798631 - 01/09/10 08:53 PM (15 years, 10 days ago)

I don't understand the need for extraction.

Look, which apples are sweeter- Red Delicious, or Granny Smith?
Sure, you can get a Granny Smith that isn't as tart. And some Red Delicious apples just aren't delicious.
But I can eat the two and tell you which is sweeter.

I don't need to extract the sugar.
I don't need to know whether it's all fructose.
I don't need to weigh the amounts of sugar in each apple on every tree in the orchard.

You guys are making this harder than it needs to be.


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