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InvisibleBaba Yaga
Psychedelic Minion

Registered: 09/13/20
Posts: 4,496
Loc: Try 'n' Error Boot Camp
Trusted Cultivator
Re: HPLC results [Re: clampconnected] * 1
    #28134973 - 01/11/23 07:52 AM (2 years, 6 days ago)

Great work. Thanks.

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OfflineYoshiTrainer
Onion tied to belt
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Registered: 04/30/22
Posts: 1,783
Loc: Castles made of sand
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Re: HPLC results [Re: Baba Yaga] * 1
    #28135033 - 01/11/23 08:53 AM (2 years, 6 days ago)

Awesome job!

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OfflineAdas
Lonely Dreamer
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Registered: 12/22/16
Posts: 5,384
Loc: Central EU
Last seen: 6 hours, 28 minutes
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Re: HPLC results [Re: YoshiTrainer] * 1
    #28135101 - 01/11/23 09:56 AM (2 years, 6 days ago)

Cool! Any additional info? Such as the time from colonization to pinning, and from pinning to harvest?

It still seems like on paper they should be less potent than Pans.

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Invisiblejoze
Male


Registered: 11/10/20
Posts: 1,056
Loc: PNW
Re: HPLC results [Re: clampconnected]
    #28135203 - 01/11/23 11:06 AM (2 years, 6 days ago)

Quote:

clampconnected said:
I am still working on this one, but figure the info that’s present now may be of interest. Psilocybe subtropicalis, mostly from Veracruz, and one from the cultivated strain ‘hoogshagenii var convexa’ or ‘semperviva’. Background images of Psilocybe subtropicalis by Alan.









Hello and thank you for sharing. I'm a fellow lab rat in Portland.

Are you working independently or with a research facility?

Could you please share more on your method?

What detector are you using?

What is your extraction method?

Have you run into any unique challenges?


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Offlineclampconnected
Researcher
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Registered: 04/07/20
Posts: 59
Loc: Portland Oregon
Last seen: 1 month, 30 days
Re: HPLC results [Re: Bismillah]
    #28136410 - 01/12/23 09:26 AM (2 years, 5 days ago)

Degradation studies are of interest. I do think it’s highly dependent on the container used, temperature, dry method, and species. Mushrooms are sponges, and if not stored properly will absorb water, which can really accelerate degradation. I personally think glass is the best container material to preserve for long periods. I’ll have to do some testing with that.

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Offlineclampconnected
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Registered: 04/07/20
Posts: 59
Loc: Portland Oregon
Last seen: 1 month, 30 days
Re: HPLC results [Re: Adas] * 1
    #28136415 - 01/12/23 09:29 AM (2 years, 5 days ago)

Most of those subtropicalis tests were ran with wild collected specimens. The one on the right was cultivated from Yoshi Amano. I’m not sure of exact times, they were mature fruits.
I think pans can be much more potent than this. I am taking pan submissions if anybody is interested in sending some material in for analysis.

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Offlineclampconnected
Researcher
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Registered: 04/07/20
Posts: 59
Loc: Portland Oregon
Last seen: 1 month, 30 days
Re: HPLC results [Re: joze] * 2
    #28136433 - 01/12/23 09:54 AM (2 years, 5 days ago)

Awesome to have some local rats!
I’m working as Tryp Labs, my independent company.
Mushrooms are desiccated prior to analysis.
Sample size is 50mg.
I extract in 1mL methanol.
Extraction is done with one hour in an ultrasonic bath around 30c, then an 18hr shake. Extracts are then filtered and a dilution is made. I’ll run the dilution for psilocybin and the undiluted extract for the minors.
I use a reverse phase system with 0.1% formic acid in water and 0.1% formic acid in acetonitrile, on a aqueous stable c18 column.
The detector is a DAD. Peak identity is by retention time and confirmed with uv spectra.
I calibrate with standards from an ISO 17034:2016 chemical supplier using a 6 point calibration curve for each analyte.
Right now challenges are coelutions that can make the results for the minor tryptamines biased high. The spectra do not always come out looking like indole, and it hinders my ability to report them in some cases. Aeruginascin is also tricky to fully separate. I need to continue working on method development. I may try going back to HILIC, or try other columns, gradient elutions, different buffer systems, it will take some
Time to resolve these issues and have a perfect method! Methanol is not the most efficient extraction solvent but it represents the mushrooms true profile the best, according to some studies. I’m not running internal standards, I need to incorporate that to begin to understand reproducibility  in my method over time.

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Invisiblejoze
Male


Registered: 11/10/20
Posts: 1,056
Loc: PNW
Re: HPLC results [Re: clampconnected]
    #28136530 - 01/12/23 11:18 AM (2 years, 5 days ago)

Quote:

clampconnected said:
Awesome to have some local rats!
I’m working as Tryp Labs, my independent company.
Mushrooms are desiccated prior to analysis.
Sample size is 50mg.
I extract in 1mL methanol.
Extraction is done with one hour in an ultrasonic bath around 30c, then an 18hr shake. Extracts are then filtered and a dilution is made. I’ll run the dilution for psilocybin and the undiluted extract for the minors.
I use a reverse phase system with 0.1% formic acid in water and 0.1% formic acid in acetonitrile, on a aqueous stable c18 column.
The detector is a DAD. Peak identity is by retention time and confirmed with uv spectra.
I calibrate with standards from an ISO 17034:2016 chemical supplier using a 6 point calibration curve for each analyte.
Right now challenges are coelutions that can make the results for the minor tryptamines biased high. The spectra do not always come out looking like indole, and it hinders my ability to report them in some cases. Aeruginascin is also tricky to fully separate. I need to continue working on method development. I may try going back to HILIC, or try other columns, gradient elutions, different buffer systems, it will take some
Time to resolve these issues and have a perfect method! Methanol is not the most efficient extraction solvent but it represents the mushrooms true profile the best, according to some studies. I’m not running internal standards, I need to incorporate that to begin to understand reproducibility  in my method over time.



Thank you for taking the time to share this information. I'm looking forward to seeing more of your results, and seeing how your method and laboratory grow over time. It will also be nice to have another scientist on the forum who can help answer chemistry questions.


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Offlineclampconnected
Researcher
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Registered: 04/07/20
Posts: 59
Loc: Portland Oregon
Last seen: 1 month, 30 days
Re: HPLC results [Re: joze] * 4
    #28676843 - 02/26/24 11:43 AM (10 months, 15 days ago)



Low resolution screen shot, but still nice none the less, of some cultivated Psilocybe caerulescens from a Georgia USA population. Nice potency on these indoor ones.
When I visited GA in 22 I tried some fresh and was taken by surprise with their strong potency. I’ve also heard they can be mild, especially from trip reports in Mexico. Environmental factors such as rain, sun, or herbivory can likely affect potency, and wild mushrooms will inherently be more variable than cultivated. But if you are considering trying your hand at cultivating this species, here’s some data to inform you it is likely well worth your efforts. I believe workman just listed some on his site not too long ago, as well as yoshi, and maybe if you’re lucky you can get a wild print from some our fellow GA or MX shroomerites.

Feel free to reply with caerulescens trip reports or link
To them.

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Invisiblejrc5
Stranger
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Registered: 02/16/23
Posts: 52
Re: HPLC results [Re: clampconnected]
    #28676854 - 02/26/24 11:56 AM (10 months, 15 days ago)

very cool..  thank you for sharing!!


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