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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: joze]
#27770685 - 05/10/22 05:18 AM (1 year, 10 months ago) |
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Quote:
joze said: Hey nektar.
I've been doing some more reading on this topic, and I think you may actually have gone too acidic now, if you're using undiluted white vinegar to extract the mushroom tissue.
Thank you. Next time I'll try white vinegar:distilled water 1:1 as my extractor, and I'll do a pH test on it.
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61]
#27770698 - 05/10/22 05:42 AM (1 year, 10 months ago) |
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That image you cited from the 1975 article talking about marquise reacting to psilocybin doesn't seem to exist anymore and only seems to be referenced on the "cops", page. Actually it's a book, and it looks creepy haha. Some old times medicine shit. I think your wasting your time on that one.
This is kinda funny tho, elrich showing a positive result on a button mushroom because of tryptophan https://www.protestkit.eu/how-to-test-psilocybin-magic-mushrooms/
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Edited by QM33 (05/10/22 05:47 AM)
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x7x_x7x
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27774468 - 05/12/22 06:35 PM (1 year, 10 months ago) |
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Quote:
nektar61 said: Test 2, Marquis Reagent
Ground dry strong cube clone (x7x) to fine with new grinder. Zeroed scale with paper before adding powder.
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sonoramo
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I just noticed this thread while looking for something completely different. Following with interest.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 2
#27775768 - 05/13/22 03:45 PM (1 year, 10 months ago) |
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Quote:
joze said: I've been doing some more reading on this topic, and I think you may actually have gone too acidic now....
Test 4, using white vinegar 50/50 with distilled water as extractor. (all my tests archived on my Journal, here.)
First I tested the pH of my 5% acetic acid white vinegar. Looks like 3. Wow. Vinegar is fucking stronger acid than I thought. No wonder it hurt like satan when I got it on a paper cut recently:
pH of my vinager diluted 50/50 with distilled water. Looks almost the same, but it's a little different. I think it's pH 4:
Since it's an acid, and the reagent I'm using (Ehrlich's) is even stronger (it's about half concentrated hydrochloric acid), I made some eye wash solution just in case. This is 30 grams of baking soda into 300 ml of distilled water. Not all of it dissolved, but it's good to have on hand, and can save eyes or skin if you get acid on them. Also I can use it to neutralize my tests (outside) after the experiment is over and before I throw them in the trash:
This is the pH of the eye wash solution. Looks like an 8, mildly alkaline:
I used the 50/50 vinegar/water solution to extract from 5 grams of BBM dried shrooms. I ground them to a powder right before the tests. Mixed 25 ml distilled water with 25 ml white vinegar. Soaked in solution for 2 hours, this time kept the jar in a water bath at 115 f.
I strained it then filtered it. Lose some in filtering. Came out of bluish. Is likely shroom blue oxidation reaction. This is bluer than previous test extractions, I surmise that the warm water bath added this time increased either the amount of psilocin / psilocybin extracted, or the enzymes that make the bluing, or both.
This is the pH of the extract. Looks like a 4, or a 4.5. Same as or slightly less acidic than the extraction solution before the extraction. (I took the solution out to test with a pipette, I did not put the litmus paper in the solution.)
I added these these amounts of extract added to fresh vials, then brought them up to 3 ml with distilled water (all measured with syringe): Vial 1 got .5 ml solution. Vial 2 got 1 ml solution. Vial 3 got 2 ml solution. Vial 4 got 3 ml solution. Vial 5 got 3 ml solution (almost, I ran out).
I added 15 drops of Ehrlich's reagent to vials 1, 2, 3, and 4. Vial 5 is a control, I added no reagent to it.
This is 5 minutes after adding reagent:
This is 10 minutes after adding reagent:
This is 20 minutes after adding reagent:
This is 30 minutes after adding reagent:
This is 40 minutes after adding reagent:
This is 50 minutes after adding reagent:
When done, I took the vials outside, dumped them into a plastic bag, and poured some of the eye wash solution on them. When it stopped fizzing, I sealed it up, doubled bagged it, and threw it out.
I wore a good filtered mask, gloves, and eye protection for this and every step of the tests. I also had a window open, and a fan blowing out the window, and a ceiling fan on.
CONCLUSION: Not much. I think that diluting the acid was a step back. I think the last test, test 3, with straight 5% acetic acid white vinegar extractor had the strongest color reaction between vials of any of the 4 tests I've done.
I promise you, aqueous acid solution is the extractor we want to use. Must have water to dissolve the psilocybin, and must have acid to make a salt of the psilocin so it dissolves well in water.
According to: https://www.swgdrug.org/Monographs/PSILOCIN.pdf (use a VPN, that's a cop site, "Scientific Working Group for the Analysis of Seized Drugs" haha):
Psilocin is only slightly soluble in water. Psilocybin is soluble in water.
Same is confirmed elsewhere online with chemical library sites.
I think we want to try MORE acidic than vinegar, but I'm not going to do it. pH 3 is my limit. But if someone who knows how to use it safely wants to try a solution of an acid that's pH 2, I think you'll get the results we seek, then can make a color chart. Even stronger acetic acid might do it, but could also probably use hydrochloric, sulfuric, or phosphoric. But I'm not fucking with any of that.
If that works better, but isn't perfect, you might want to try again adding a small amount of ferric chloride, to get the effect of Keller's Reagent too. That reacts with the enzymes we're testing for.
So I've done 4 tests, I think I've found some useful info, and I'm passing the torch. -- EDIT: after looking again at the results, I think the tests at 20 minutes may have been useful. It's hard to tell because there is just the bluing with the dilution, but at 20 min it looks like the ones with more drugs in it looks more dark brown. Might be on to something here. What do you think?
If someone has some weak shrooms and some strong shrooms and wants to repeat this test with two extraction solutions, that might be good. I'm getting burnt out on breathing acid fumes.
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Edited by nektar61 (05/14/22 11:39 AM)
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nektar61
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Quote:
SpectreOfCommunism said: ...Hofmann reagent ...is made with DMAB, sulfuric acid, and ferric chloride.
Anyone know how much ferric chloride to how much of the other ingredients? Can't find it anywhere.
A lot of people sell Hofmann's, but no one seems to want to give up the recipe for the secret sauce. (not surprising)
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27787387 - 05/21/22 07:55 PM (1 year, 10 months ago) |
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TEST 5. BRING ON THE HEAVY METAL (Iron that is.... ferric chloride solution 40% - printed circuit board etchant) -- I think this one is showing promise. Just need to tweak the amounts. Added ferric chloride 40% to Ehrlich's reagent, sort of made an Ehrlich's / Hoffman cross.
I ground 2.4 grams of BBM clone shrooms (dry). Soaked in 40 ml white vinegar. (comes as 5% acid, 95% water).
Soaked for 80 minutes. Stirred vigorously every 20 min, plus at start and end.
Strained, then filtered. Got this:
This is a weaker solution than I made last time, which I think is good. Last time was so strong it had more bluing, which could interfere with reading color-based test results.
Here's what went into each vial:
Vial 1: 3 ml shroom extract. Vial 2: 3 ml shroom extract. Vial 3: 3 ml shroom extract. Vial 4: 3 ml shroom extract.
(This is pic before adding some different amounts of shroom extract to each of the vials after #4)
Added 10 ml of Ehrlich's reagent to each vial in Vials 1-4. Not much reaction.
Added tiny amounts of ferric chloride solution to Vials 1-4.
Vial 1: Added .2 ml ferric chloride solution. Vial 2: Added .4 ml ferric chloride solution. Vial 3: Added .75 ml ferric chloride solution. Vial 4: Added 1 ml ferric chloride solution.
Got INSTANT reaction of dark green on all, could not tell them apart:
Wanted to see if it was just a reaction to the vinegar. Poured shroom extract from Vial 5 into Vial 6 (which then had 6 ml extract).
Washed out Vial 5 with distilled water. Filled it with just 3 ml of white vinegar. Added ferric chloride solution, .25 ml. Did not turn green (shown in above photo). Turned straw / urine yellowish.
For Vials 6-9, adjusted to these amounts of shroom extract:
Vial 6: 6 ml shroom extract. Vial 7: .5 ml shroom extract, adjusted with distilled water up to 3 ml. Vial 8: 1 ml shroom extract, adjusted with distilled water up to 3 ml. Vial 9: 2 ml shroom extract, adjusted with distilled water up to 3 ml.
Then added the Iron: Vial 6: Added 3 drops ferric chloride solution. Vial 7: Added 1 drop ferric chloride solution. Vial 8: Added 5 drops ferric chloride solution. Vial 9: Added 2 drops ferric chloride solution.
I was trying different things to try to get SOME visible variation.
It's very hard to see, but I think that the ones with less had slightly lighter reaction. Hard to tell but under bright light, could see it. Couldn't photograph well though. Below pic is, left to right, Vials 6-9.
As you can see if you look closely, Vial 7 is slightly lighter green than the others. It had the least shroom extract and the least ferric chloride:
CONCLUSION: the VERY fast VERY abrupt reaction is going in the right direction. I think next step is to make Ehrlich's reagent with much smaller amounts of ferric chloride 40% mixed in. Maybe start with 1 drop per 3 ml and see, then go down or up from there to get see if we can get a quantitative reaction.
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Edited by nektar61 (05/21/22 08:03 PM)
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7Suns
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27792925 - 05/25/22 08:40 PM (1 year, 9 months ago) |
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Not sure if this is anything useful but have you guys also tried fluorescing the different concentrations under black light ? I’ve seen something about using black light to check tartaric powder extracts posted by achuma I believe
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: 7Suns]
#27793022 - 05/25/22 09:50 PM (1 year, 9 months ago) |
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Quote:
7Suns said: Not sure if this is anything useful but have you guys also tried fluorescing the different concentrations under black light ? I’ve seen something about using black light to check tartaric powder extracts posted by achuma I believe
I tried both long UV (black light) and short (mineral light, the kind that requires goggles) on the last batch I did, no florescence, but I did it after I added the reagents. Will try before on next.
A lot of things can respond to UV though.
When you say "tartaric powder extracts" do you mean LSD tartrate, or ergotamine tartrate? Or something else.
I've heard of UV light being used to separate out levo and dextro LSD in a chromo column. Not sure if that applies here.
One thing I do plan to do if I, or someone else here, gets something that works for quantitative visual reagent tests: test it on non-psychedelic shrooms to make sure we're not getting a hit on something other than the psilo alkeloids.
Thank you.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: 7Suns]
#27793044 - 05/25/22 10:11 PM (1 year, 9 months ago) |
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Quote:
7Suns said: I’ve seen something about using black light to check tartaric powder extracts posted by achuma I believe
OK, you mean this, right?: https://www.shroomery.org/forums/showflat.php/Number/22960162
I don't think that's florescence. That said, keep them coming. I'm interested in ANYTHING that will help figure this out.
Real florescence is almost shocking, and gets your attention even at low levels. I think what they were seeing is reflection of the visible component of their UV light. They also don't even say what type of UV it is. So probably black light, not what they probably want for this. I tried that and a "real" UV light, and didn't see anything. As I said, was after tests, and will repeat again before tests. (but then not test that tube, as UV can degrade chemicals.)
I don't think achuma got pure psilo alkaloids. They say it's 1/10 the weight of dried shrooms. Should be more in the magnitude of 1/100th.
They say it's active, but probably also contains a lot of sugars, proteins, and other non-actives that will mess with the results. I'm still going though the thread, but I think I agree with this: https://www.shroomery.org/forums/showflat.php/Number/22964630#22964630
Quote:
Regarding your UV "fluorescence" I believe what your observing is mere reflection of your blacklight. Psilocybin and psilocin lacks the conjugated double bond found in lysergic acid derivatives that emit fluorescence (opposed to just reflecting it).
This is what milk looks like under blacklight, it's not glowing, just reflecting.
Also see this thread for more fluorescence of compounds that contain conjugated double bonds (again more lysergic acid derivatives)
Edit: you have demonstrated that light can be reflected, you still lack proof of actual fluorescence
and this; https://www.shroomery.org/forums/showflat.php/Number/22965121#22965121
Quote:
What is being stated in that article is if you excite psilocybin with a 267nM light it will fluoresce at 335nM (which cannot be detected by the human eye) and if psilocin is excited with a 260nM light it will fluoresce at 312nM (which cannot be detected by the human eye). The human eye can only detect wavelengths from 390 to 700 nM
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7Suns
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27793308 - 05/26/22 06:34 AM (1 year, 9 months ago) |
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Yes that’s the link, yea I wasn’t sure if it was useful or not
-------------------- Following the lost souls as they transcend the seven dimensions, through playful dancing, like wandering shadows in the heat of a mirage 7suns grain sac hack 7suns straw log hack 7suns official OTC antibiotic agar recipe 7suns DIY filter bags 7suns cheap AF flowhood 7suns cold weather fruit chamber
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: 7Suns] 1
#27797129 - 05/28/22 11:48 PM (1 year, 9 months ago) |
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TEST 6 modified Hofmann's Reagent, plus UV tests
I ground fine 2.5 grams of dried BBM clone. Added 40 ml 5% acid white vinegar. Shook at start and every 15 min and at end for one hour. Strained, then filtered twice with coffee filter. Result:
Here's the Ehrlich's Reagent I'm using:
Here's a fresh 3 ml bottle after adding 1/10 of 1 ml of ferric chloride, showing the slight color change (just dilution). I'm making a very modified variation (almost a distant cousin) of Hofmann's reagent:
Here's the solution I added, and a syringe showing trying to measure just .1 ml. Was hard, air bubbles, and end of syringe neck, but I think I got it after this to just 1/10 ml and added to the fresh bottle above.
Below, Vial #7, shows 3 ml shroom extract plus 1 drop ferric chloride solution only, no Ehrlich's Reagent added. Has immediate dark green reaction like test 5 with ferric / Ehrlich's. I believe now that the tests last time are just showing a reaction with something in the shrooms plus iron only. No reagent needed, so test is not conclusive.
It may even be measuring proteins or sugars. Did not show last time with just ferric and vinegar, is something in shrooms.
NOTE: To anyone who does these tests, if you get something you think is conclusive, then also test on non-psychedelic mushrooms to rule that out.
The other vials here are 2 ml shroom / vinegar extract with nothing added:
Other tests I did as long as I had everything out.
2 ml shroom / vinegar extract. 3 drops modified Hofmann's:
2 ml shroom / vinegar extract. 5 drops modified Hofmann's:
2 ml shroom / vinegar extract. 2 drops modified Hofmann's:
.5 ml shroom / vinegar extract. 1.5 ml distilled water. 2 drops modified Hofmann's:
I'm pretty sure these are just showing dilution of the reaction of vinegar and ferric. So this probably isn't detecting active alkaloids.
UV LIGHT: I had a little leftover vinegar / shroom solution so I tried UV light.
These are the lights I used. Cheap long wave black light flashlight on the left, and a better shortwave UV light for rock collecting on the right. If you do these tests, the shortwave requires eye protection.
Here's the unused shroom / vinegar extract illuminated in the dark with cheap long wave black light;
Here's the solution illuminated in the dark with better short wave UV light;
Neither of these are fluorescing. There is a little reaction with the glass on the long wave one. Other than that the liquid is just reflecting a little bit of the viable component of these UV lights
CONCLUSION: No conclusion, but science includes showing what doesn't work. I did show some things that might not need to be tried again, or someone can repeat them to show they're repeatable. But they seemed pretty conclusively non-reactive to me.
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27798661 - 05/30/22 06:07 AM (1 year, 9 months ago) |
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“I have not failed. I've just found 10,000 ways that won't work.”
Edison
-------------------- OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum... There are known knowns, there are known unknowns, there are also unknown unknowns. With great privilege comes great responsibility. Quantom Qups PROOF AND Soft Drops Turn your Swab to a Syringe and Syringe to Multiple Syringes! No Pours (QuantomStyal)Magic Fruit Leather DMT for IandI
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nerdintheshell] 1
#27861451 - 07/13/22 04:56 PM (1 year, 8 months ago) |
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Old thread, but if anyone is still interested in TLC there was a post recently describing relatively simple process for comparison testing that might be worth checking out.https://www.shroomery.org/forums/showflat.php/Number/27830910#27830910
Quote:
Notes on sample preparation and procedure for the above shown TLC]
100mg of carefully weighed, blended/homogenized mushroom powder from each sample was added to a small beaker, 5ml of methanol was added over top while also washing down the sides of the beaker. The beakers were covered with plastic wrap and secured with rubber bands then transferred to an ultrasonic bath in hot tap water, sonicated for 8 minutes with heat, 5 minute rest, followed by another 8 minute sonication with heat. Samples were allowed to settle for 10 minutes but no filtration, decanting, or centrifuging was done. A calibrated pipette was used to transfer 10µg of each sample to the chromatography sheet. Sheet was developed in a sealed glass chamber saturated with eluent consisting of 100 parts methanol: 1.5 parts ammonia. After development the sheet was allowed to dry while undergoing ultraviolet examination, then freshly mixed Ehrlich's reagent was applied by spray bottle until the sheet was saturated, followed by drying with a heat gun. Sheet was periodically photographed every 5 minutes until colors reached full development (maximum brightness).
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MariaTruthSeeker
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt]
#27976801 - 10/01/22 03:14 PM (1 year, 5 months ago) |
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Quote:
Icyurmt said: Old thread, but if anyone is still interested in TLC there was a post recently describing relatively simple process for comparison testing that might be worth checking out.https://www.shroomery.org/forums/showflat.php/Number/27830910#27830910
Quote:
Notes on sample preparation and procedure for the above shown TLC]
100mg of carefully weighed, blended/homogenized mushroom powder from each sample was added to a small beaker, 5ml of methanol was added over top while also washing down the sides of the beaker. The beakers were covered with plastic wrap and secured with rubber bands then transferred to an ultrasonic bath in hot tap water, sonicated for 8 minutes with heat, 5 minute rest, followed by another 8 minute sonication with heat. Samples were allowed to settle for 10 minutes but no filtration, decanting, or centrifuging was done. A calibrated pipette was used to transfer 10µg of each sample to the chromatography sheet. Sheet was developed in a sealed glass chamber saturated with eluent consisting of 100 parts methanol: 1.5 parts ammonia. After development the sheet was allowed to dry while undergoing ultraviolet examination, then freshly mixed Ehrlich's reagent was applied by spray bottle until the sheet was saturated, followed by drying with a heat gun. Sheet was periodically photographed every 5 minutes until colors reached full development (maximum brightness).
Good reference. I have been wondering which would be better, using TLC or a Miraculux like solution as described in this thread. I think the TLC solution described above could be simplified a little more, a single TLC sheet could be cut into several small sheets and developed in a standard 16oz mason jar. As you can see from the picture in that above thread, visualization is quite good with Ehrlichs and you can easily see both psilocybin and psilocin separately which is a big advantage over the Miraculux/reagent solution. Psilocin is significantly more potent by weight and the Miraculux type solutions may not be accurate for high psilocin species (like Pan Cyans). If someone came up with an exact procedure others could replicate, then you could measure the area of the spots using Gimp or other photo editing software and build out a table/graph of spot sizes to actual percentage for each alkaloid. I'm not sure how doable that would really be though, as spot size alone may not be adequate, maybe the darkness/color matters too? The other problem is everyone's extraction would have to be the same, while sonication is probably the best known extraction method, everyone would have to do it the same way with the same wattage, etc, and not many would own a sonicator so it wouldn't be cheap/easy for many people. The standard solution would probably have to use a free extraction method (maybe just soak for 24 hours in solvent with periodic shaking?). Likewise, to be reproducible, everyone would need a proper calibrated pipette, they aren't that expensive but still this is another tool people would have to buy and another barrier to accessibility.
I suppose just doing "comparisons" like was done between the Penis Envy and Pan Cyan to get a good sense of the potency difference is probably adequate for many people. You could use this to compare old stored samples vs. freshly grown/dried samples to see how much potency has been lost, or to compare one species to another, etc.
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Suckatshrooms
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#28009921 - 10/21/22 08:15 PM (1 year, 5 months ago) |
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HCL is nasty. There’s almost always a better solution. Sulfuric acid is great. It’s cheap, easy to find locally and stays in solution so it’s much safer. HCL is a gas so respirators are needed, much like it’s basic brother, ammonium hydroxide. HCL outside a fume hood will rust everything in your lab as well.
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hjalmar
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Re: DIY Psilocybin Content Percentage Detection [Re: Suckatshrooms]
#28036538 - 11/06/22 01:33 PM (1 year, 4 months ago) |
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Quote:
Suckatshrooms said: Sulfuric acid is great. It’s cheap, easy to find locally and stays in solution so it’s much safer.
Depends where you live. In the EU since a few year ago the sale of sulfuric acid over 15% to private individuals has been banned. They're selling 10% H2SO4 now as drain cleaner, I doubt if it will even do that, only licensed professionals are allowed to handle to concentrated stuff now (i.e. you pay 150 euros instead of 15).
Ayway shouldn't be a problem to handle muriatic acid (the 25% HCl the hardware store stocks) as long as you take the necessary precautions.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: hjalmar]
#28039162 - 11/07/22 08:13 PM (1 year, 4 months ago) |
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Quote:
hjalmar said:
Quote:
Suckatshrooms said: Sulfuric acid is great. It’s cheap, easy to find locally and stays in solution so it’s much safer.
Depends where you live. In the EU since a few year ago the sale of sulfuric acid over 15% to private individuals has been banned. They're selling 10% H2SO4 now as drain cleaner, I doubt if it will even do that, only licensed professionals are allowed to handle to concentrated stuff now (i.e. you pay 150 euros instead of 15).
Ayway shouldn't be a problem to handle muriatic acid (the 25% HCl the hardware store stocks) as long as you take the necessary precautions.
5% acetic acid vinegar, can you get that there? It's pretty low Ph, and I was trying to use it for my extractions here.
I think it might work.
-------------------- -NEW? Start here.
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