| Home | Community | Message Board |
|
You are not signed in. Sign In New Account | Forum Index Search Posts Trusted Vendors Highlights Galleries FAQ User List Chat Store Random Growery » |

This site includes paid links. Please support our sponsors.
|
| |||||||
|
| |||||||
|
Master Exploder! Registered: 11/15/07 Posts: 1,237 Loc: Las Vegas Last seen: 1 year, 5 months |
| ||||||
|
I havent looked into everything yet but I did look at used equipment, even the GCMS was affordable. New equipment is another story. I have to wait for the money, then do the research. I dont see why this couldnt be done in a homemade lab under sterile conditions? I would be using professional grade lab equipment that would make a cave suitable. The thing Im most worried about is not being able to obtain the right chemicals if they are needed.
Im not sure if you understand me correctly but I want to use this machine to make a more potent cubensis. If I have to make an isolate of it I will. They can be stored almost indefinitely. I really dont want to have many people counting on me. Im not being selfish or mean but I just dont want the pressure,I want to enjoy what Im doing. I would be pressured for time and I dont like that idea.For the project, I will take notes and pictures. I will post them in the forum when I get setup. My guess is that all cubensis sub strains(except PE) are going to vary mushroom to mushroom,flush by flush and substrain is not gonna matter. I think it will come down to genetics on potency and genetics is a game of chance. -------------------- Cloneufc's easy Penis Envy spore tek! Cloneufc's easy Cloning tek Cloneufc's 360 degree LC Cloneufc's No foil jar tek! Cloneufc's Breaking up spores Edited by Cloneufc (03/02/10 03:45 AM)
| |||||||
|
Ascension Energy & Alien UFOs Registered: 05/12/07 Posts: 71,179 Loc: The Inexpressible... Last seen: 3 hours, 18 minutes |
| ||||||
|
All good points, you sure know your stuff....
But would you consider someone paying you a small fee to have you analyze their shrooms? I think a lot of people would be interested in it, as long as you provide some good results. And even a couple samples of PE with a couple samples of some Control cubenis (i.e. "B+") would help this project tremendously! As for chemicals, is that required? I thought you could just use the material straight-up, as long as its small enough....any GCMS experts here? I just hope you get the money, because I would pay up to $20 to have shrooms analyzed by your machine, as long as I saw some trials on your mushrooms first. There May be hope ~ LogicaL Chaos ~
| |||||||
|
Registered: 12/25/09 Posts: 1,875 Last seen: 6 years, 11 months |
| ||||||
|
I like the way you guys are going with this. Good work.
| |||||||
|
Thread Killer Registered: 08/05/05 Posts: 608 Loc: west of a white |
| ||||||
Quote: I've done a fair amount of GCMS, but I wouldn't call myself an expert. You don't need any chemicals other than the extraction solvent and the inert gasses. The solvent used depends on the column in the machine and the analyte; I usually used methylene chloride, but for the solvent to be appropriate the analyte must also be soluble in it. It's been a while since I looked up psilocybin solubilities. Water can't be injected into a GC without messing it up, it will clog the column. A GC will constantly have a stream of heated inert gas passing through it, so a tank of such a gas would be necessary (and easily found at your local welding/gas supplier). Tiny amounts of solvent are used, so I'm sure you could get any that you would need and still fall under the radar. When injecting into a GC, you use a microliter syringe. The amount of solvent injected is minuscule, the amount of analyte you inject into the machine is incredibly small. About sending Clone samples to analyze, I think that is a bad idea. It is impossible to receive a package without giving out an address, and that wouldn't be good for all of us to have his address especially if he is doing something that illegal. It would be cool, but an impractical aspiration. -------------------- "Now ether was substituted for chloroform, and the difference of their phenomena noted, and now some other exhilarant, in the form of an opiate or stimulant, was the instrument of my experiments, until I had run through the whole gamut of queer agents within my reach..." I can do everything!!
| |||||||
|
liberator Registered: 08/06/08 Posts: 4,204 Last seen: 4 years, 6 months |
| ||||||
|
Indeed, don't send out your address if you're undertaking such a project as this. Although, mass producing mushrooms is probably way more illegal than chemically analyzing mushrooms, and people send out their addresses all the time to give and receive prints. As long as it's from a trusted member who has been around for a fair amount of time and has good trade ratings, I think it should be safe.
For a solvent, methanol seems to be the most reliable.
| |||||||
|
Master Exploder! Registered: 11/15/07 Posts: 1,237 Loc: Las Vegas Last seen: 1 year, 5 months |
| ||||||
|
Thats the main reasons why I will not do anything like that through the mail. Posting results and testing the results will suffice. Even if I was caught and I had a GCMS, I would probably just get a fine. I mean how many people have these in theirs homes? With a good lawyer, I could claim medical research and get off with a fine for practicing without a schedule 1 license.
-------------------- Cloneufc's easy Penis Envy spore tek! Cloneufc's easy Cloning tek Cloneufc's 360 degree LC Cloneufc's No foil jar tek! Cloneufc's Breaking up spores
| |||||||
|
Ascension Energy & Alien UFOs Registered: 05/12/07 Posts: 71,179 Loc: The Inexpressible... Last seen: 3 hours, 18 minutes |
| ||||||
|
What a bummer, but you do have a good reason for not wanting to (ya know, legal concerns).
How about this: WE grow some mushrooms, take a spore print, send you the print and you fruit them. Hows that? No legal problems and I don't mind giving out my address to a few people as long as they are responsible with it and don't black mail (small pun intended) me later for growing mushrooms. Would you consider that Cloneufc? I'd still pay like a small fee to help with growing costs. I could even send some powdered brown rice flour and vermicilite with it to reduce costs on you. That would still be good for this project, but that means *you* will be doing most the work. With you being the only one with a GCMS machine, this project could get some reliable and interesting results... And since you will be recieving spores from all over the world, you can clone the most potent mushroom you grew and tested for yourself and start a business. Pretty good deal right? Would you consider my idea? ~ LogicaL Chaos ~ Edited by LogicaL Chaos (03/03/10 07:05 PM)
| |||||||
|
Mycologist in Trainning Registered: 12/21/09 Posts: 4,622 |
| ||||||
|
since each print has millions of different genetic's, i dont think that would do anything at all to actually tell how potent YOUR shrooms are...every batch will be different.
| |||||||
|
Ascension Energy & Alien UFOs Registered: 05/12/07 Posts: 71,179 Loc: The Inexpressible... Last seen: 3 hours, 18 minutes |
| ||||||
|
Well then he would have to do multi-batches, powder ALL the dried shrooms into one large collection, and then take a sample from that.
Again, maybe too much work for the Clone Man, but he's willing, I'm willing, ya know? ~ LogicaL Chaos ~
| |||||||
|
Thread Killer Registered: 08/05/05 Posts: 608 Loc: west of a white |
| ||||||
Quote: Growing mushrooms is often less offensive than chemically purifying them/or altering them by extraction. One person would be seen as a "cultivator" the other would be seen as "a drug cook with a clandestine lab." It depends on how bad they want to get you. I'm sure the DEA would love to take down somebody managing to have any success. Sometimes methanol is too polar for GC, it would have to be a column designed for its use. GC and IR have to be my favorite analytical methods, I would love to have those machines. I've been too afraid of trouble fixing old machinery, or the cost of upkeep. Then I'd want nmr, and hplc, fluorescence, pressure columns, sequencers... gawww
-------------------- "Now ether was substituted for chloroform, and the difference of their phenomena noted, and now some other exhilarant, in the form of an opiate or stimulant, was the instrument of my experiments, until I had run through the whole gamut of queer agents within my reach..." I can do everything!!
| |||||||
|
liberator Registered: 08/06/08 Posts: 4,204 Last seen: 4 years, 6 months |
| ||||||
Quote: I'm not sure that's true, and I've heard plenty of "cultivators" demonized as mad scientists who were operating a drug lab, even though nothing they were doing was remotely as technical as anything that goes on in a lab. Cultivating has way more risk of getting found out as well, because there is always a possibility that someone you give the mushies to will squeal, but obviously analyzing mushrooms has no customers or other parties involved that could lead to you getting busted except the members of this community. Like I said, as long as the people sending samples are well trusted. I also don't think we should rule out the use of plain alcohol or methanol extractions for analysis, so as to keep the sample population at least a bit larger than one person in their home with a machine. We should work on coming up with a protocol for how we can test an extraction method for it's reliability against other extraction methods so we can start getting lots of other members involved. I think if we have a set protocol that we know works, and lay it out like a tek, more people will be inclined to provide samples. For example, if the alcohol is pure, nothing other than psilocybin should be extracted into that alcohol, correct, at least in theory? If this is the case, we should grow out a clone or an isolate, so we know the genetics of the flush will be absolutely uniform, and do several extraction trials with several different samples from each flush. If the method is consistent and effective, every trial should come up with the same amount of extract, correct? Then the amount extracted can be weighed and then the original wet weight of the mushrooms divided by that number to figure out the percentage of psilocin/psilocybin content. Does this sound like a start?
| |||||||
|
Ascension Energy & Alien UFOs Registered: 05/12/07 Posts: 71,179 Loc: The Inexpressible... Last seen: 3 hours, 18 minutes |
| ||||||
|
On the subject of extraction Liber, alcohol and methanol extractions aren't exactly that clean and precise....
And I learned that from personal extractions I've done myself (see my journal) and from this: Here's what chemistry-expert FastFred (should he have the title "Trusted Chemist"?) Said from my own thread I made in the "Chemistry & Pharamcology" forum called Regarding Psilocybin Extraction: Is This Statement True? (Note: questions with " > " were original written by me, and answered by FF.) Quote: So, as you can see from his answers, alcohol and methanol extractions are very crude. The resulting extract from mushrooms might even have a higher ratio of non-actives than mycelium (just a logical theory I have, since mushrooms are more complex organically than mycelium). But, as long as its consistent between ALL mushrooms, meaning some sub-strains (like PE) won't have *more* of these non-active organic protiens and carbohydrates than others, the results should give us a rough idea of how much actives they have. The best part about is that its simple and pretty expensive, and you don't need a lot of tools/utentiles to do it (i.e. you don't need a fully-stocked lab), which means nearly anyone who can grow shrooms can do these simple extractions.... I think this is the best bet, unless someone here wants to volenteer to do a full-on, psilocybin-only extract and has a fully-equiped lab to do so, which I seriously doubt (but you never know). Keepin' it simple has its downsides.. ~ LogicaL Chaos ~ Edited by LogicaL Chaos (03/04/10 09:19 PM)
| |||||||
|
liberator Registered: 08/06/08 Posts: 4,204 Last seen: 4 years, 6 months |
| ||||||
|
Interesting, for some reason I thought A/B extractions were completely out for psilocybin extraction. I guess it does make sense though that it would work better than just one solvent. If I can get my hands on some of the chemicals involved, I would definitely be willing to try out an A/B extraction. I wonder how NOKHTA's extractions turned out...
| |||||||
|
liberator Registered: 08/06/08 Posts: 4,204 Last seen: 4 years, 6 months |
| ||||||
|
Oh yea, forgot to mention, I've posted this project across two of other mushroom cultivation forums.
They can be found here: http://forums.mycotopia.net/fung and here: http://shroomotopia.net/index.ph If anyone knows of any other mushroom cultivation or drug chemistry forums, feel free to post them there as well. I'll probably post them at the dmt nexus once I get posting privileges there.
| |||||||
|
Stranger Registered: 07/23/09 Posts: 521 Last seen: 8 months, 28 days |
| ||||||
|
My extractions didn't turn out so well lol. I messed up a bunch of stuff. I'm waiting to get some ph calibration liquid for my new ph pen before I go at it again.
-------------------- Most recent contest posts: inoculation Ready for spawning Substrate done colonizing Pins Pins #2 First flush
| |||||||
|
liberator Registered: 08/06/08 Posts: 4,204 Last seen: 4 years, 6 months |
| ||||||
|
Damn, that stinks. Hopefully you'll get better results next time, and also hopefully others will try the methods out as well.
| |||||||
|
Master Exploder! Registered: 11/15/07 Posts: 1,237 Loc: Las Vegas Last seen: 1 year, 5 months |
| ||||||
|
I just found this article on how to do it. It took awhile because the study was done in Holland and the PDF file was in Dutch. I cant read dutch so I had to find a google translator to convert it to English. I haven't read it yet but Im excited to read it. Original file here: http://www.lycaeum.org/mv/TMC/02
Page 1 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 1 of 17 Southern Inspection Services Division Multiplex Signaling Products Sector: Signaling Lab Psilocybin and psilocin IN Shrooms LFJ van de Laak, JCA van der Wielen and PH in 't Veld Inspection Service of Goods PO Box 2280 5202 CG 's-Hertogenbosch Tel: 040-29 11 500 Fax: 040-29 11 600 E-mail: @ Leo.van.de.laak kvw.nl Date: July 2002 Project identification or report SAZD/01/30/21 Copyright 2002 1 1 Copyright 2002 Southern Food Inspection / Detection Division. Reproduction is authorized by written permission of the author (s) and source ________________________________ Page 2 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 2 of 17 CONTENTS TABLE OF CONTENTS 2 SUMMARY 3 1 INTRODUCTION 4 1.1 General 4 1.2 Shrooms 4 1.3 Research Techniques 5 2 MATERIALS AND METHODS 5 2.1 Methodology 5 2.1.1 Chemicals 5 2.1.2 Sample Preparation (sample preparation) 6 2.1.3 Standard and sample solutions 6 2.1.4 LC 6 2.1.5 Robustness Study 7 2.1.6 Validation Study 7 2.2 Monitoring Shrooms 7 3 RESULTS 8 3.1 Methodology 8 3.2 Monitoring Project 8 4 DISCUSSION 9 4.1 Methodology 9 4.2 Monitoring Project 9 5 CONCLUSION 10 6 THANKS 11 7 LITERATURE 11 8 ANNEXES 12 8.1 Chromatograms 1 and 2 12 8.2 Tables 1 and 2 13 8.3 Charts 1, 2 and 3 15 ________________________________ Page 3 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 3 of 17 SUMMARY For the project "Maintaining and monitoring the smartshop industry" was a method for the determination of the hallucinogenic substance psilocybin and psilocin in mushrooms developed. Because of the simple sample preparation and good separations were an LC method Muss Hoff [7] as a starting point for research. By conditioning problems of the LC system was finally chosen and psilocybin psilocin through separate isocratic analysis to determine. Both assessments were subject to a robust research which examined factors (vibration time and temperature in the ultrasonic bath and the percentage of acetonitrile in the mobile phase) within the margins were found to be not critical. The validation study showed that the analysis with Recovery and quantification limits of respectively 93.4% and 0.009% for psilocybin and of 95.8% and 0.005% for psilocin good results. Using this analysis were validated by the smart shops planned monitoring samples (mainly fresh and dried mushrooms) and a mushroom grower sampled fresh mushrooms investigated. Some Fresh Panaeolus cyanescensmonsters was a much higher total psilocin content found, than in the smart shops sampled dried variety (2.5% versus 1.5%). The form in which the psyco-active substance is present (psilocybin or psilocin) is this mushroom-type completely arbitrary. This applies to both fresh and dried the samples. The Psilocybe cubensis mexican, whether or not dried, had a total psilocin between 0.5 and 0.9 %, This value was for the fresh Psilocybe cubensis thai only marginally below 0.5 - 0.7%. In the Psilocybe Tampanensis (truffle), a total of about 0.3% psilocin measured. The measured rates of psilocybin and psilocin in dried mushrooms or not be correspond to the values in the literature are listed. Only the occasional but very high psilocybin and psilocin levels (and therefore the total psilocinegehalte) in freshly picked Panaeolus cyanescens are striking. The investigation also revealed that the psycho-active substances in fresh mushrooms, except in the Psilocybe Tampanensis, much less stable than in the dried form. The hallucinogenic effect is due to degradation of the two very different psychoactive substances. KEYWORDS: psilocybin, psilocin, smart shop, mushrooms, indole alkaloids ________________________________ Page 4 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 4 of 17 1. INTRODUCTION 1.1 General Shrooms (mushrooms with hallucinogenic properties) are among the oldest known strip means. Trade (through smart shop) and the use of hallucinogenic mushrooms since 1995 strongly challenged in the Netherlands. Lately, however, the use of mushrooms all over again its peak to be [7.15]. The active (mind-altering) substances psilocin and psilocybin mushrooms are. Both substances are tier 1 of the Opium Act and are therefore regarded as hard drugs. Growing psilocybin and psilocin-containing mushrooms with the intention to prepare, edit and sell (etc.) is a criminal act under Article 10a of the Opium and is punishable. Trade in fresh mushrooms and growing trays to grow magic mushrooms is not prohibited, these activities fall under the Commodities Act. In a study of the risks that these mushrooms to public health, came The Ministry of Health Assessment and Monitoring Coordination Committee established new drugs (CAM) concluded that this must be assessed as low. Reactions Acute toxicity limited to panic and anxiety attacks. In chronic toxicity can most of the action called flashbacks. There is also no physical or mental dependence [7.15]. In the section of Signaling KvW South in the context of the project "Preservation and monitoring in smartshop industry "developed an analytical method, which limits psilocin and psilocybin in magic mushrooms can be determined. In 2001, a study of mushrooms in the smart shop branch out. In two periods of 3 months (March to May and September to November), there are 20 samples twice mushrooms for a monitoring sampled. In these samples were the newly developed method of psilocybin and psilocin levels determined. The purpose of this monitoring was to a clear picture of the current situation in the world smartshop occurs (eg fresh / dried mushrooms, psilocybin and psilocin levels occurring in mushrooms, types mushrooms, etc.). The results of this market survey in the future as a basis at any higher levels of psilocin and psilocybin in magic mushrooms, obtained by Applying changed farming methods, to identify. 1.2. Shrooms In recent years, the mushrooms as "natural" eco-drugs, partly due to the rise of the house culture is very popular. In the Netherlands the following mushrooms are most used: a) Psilocybe cubensis Stropharia cubensis or ('Mexican' or 'Thai' magic mushrooms). b) Psilocybe semilanceata (indigenous pointy bald head). c) Panaeolus cyanescens cyanescens or Copelandia ("Hawaiian" or "Balinese" mushrooms). d) Psilocybe Tampanensis (truffles), root-like mass of hardened mycelium with a dry matter content of about 30 percent. The active substance in various mushroom species varies from almost nil to 15 mg / g dry weight. The fresh mushrooms, this is a strength factor of 10 lower, as a fresh mushroom (excluding truffles) for approximately 90% water. The mushrooms may be either fresh or dried, often in combination with other foods, consumed. As usual dose for a mushroom trip is a quantity of 8 to 10 mg of psilocybin / psilocin. Average mushroom contains approximately 90% water, 3% protein and additionally small nitrogen-containing compounds, carbohydrates, fats, salts, minerals and vitamins. Especially the proteins and nitrogen compounds play a major role in mushroom poisoning. Many of the fungi occurring toxins ________________________________ Page 5 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 5 of 17 amino acids (building blocks of proteins) or from the amino acids formed. This includes the indole alkaloids psilocybin and psilocin, which are formed from the amino acid tryptophan. Psilocybin and psilocin are organic compounds, the heterocyclic built nitrogen atom is responsible for basic properties. These compounds are mainly in plants and have specific effects on the nervous system. The structure formulas of psilocybin and psilocin are very similar. Psilocybin (4-fosforyloxy- N, N-dimethyltryptamine) can be regarded as the phosphoric ester of psilocin (4-hydroxy-N, N- dimethyltryptamine). When psilocybin is ingested by the enzyme alkaline phosphatase rapidly and completely converted into psilocin. This is the psychoactive effect completely due to psilocin. + H 3 PO 4 ============> psilocybin enzyme + H 2 O psilocin The working volume of 'toxin' in a mushroom should better be displayed in a total psilocin content. A better comparison between the various mushroom species for the hallucinogenic effects is also possible. 1.3. Research Techniques Various techniques have been applied to psilocybin and psilocin in magic mushrooms to identify. Besides the paper chromatography [7.1] and thin layer chromatography [7.2 and 7.3] were later instrumental techniques developed to quantitatively analyze indole alkaloids such as: conventional column chromatography with UV-spectrometry and colorimetry, gas chromatography (GC) and mass spectrometry (GC-MS). These instrumental techniques are however, quite time consuming. In GC and GC-MS techniques [7.4], both compounds also be derivatized, and degradation of the components can not be determined can be ruled out. Currently, the LC method (reversed phase) combined with UV, fluorescence or electrochemical detection [7.2, 7.5, 7.6 and 7.9] more frequently used for the quantitative determination of psilocybin and psilocin in magic mushrooms. The advantage of the LC technique is very simple extraction of both substances from the crushed dried sample material methanol [7.5 and 7.7]. An additional advantage is that the derivatized extracts are not required to and readily lend themselves to research (analysis). Because of the simple sample preparation, separations and good results was a LC method (reversed phase, UV detection at 266 nm) [7.7] as the basis for research. 2 MATERIALS AND METHODS 2.1 Methodology 2.1.1 Chemicals • Standard psilocybin A: Sandoz Switzerland, received from Dr. WG van der Sluis of Faculty of Pharmacy, Department of Pharmacognosy, University of Utrecht (purity 99%). • Psilocin standard: Prepare the standard using a psilocybin by Dr. WG van der Sluis been handed procedure involving psilocybin quickly and completely using the enzyme alkaline phophatase can be converted into psilocin .. ________________________________ Page 6 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 6 of 17 • Standard psilocybin B: Alltech / Applied Science USA. (Ampoules containing 100 micrograms / ml in methanol). • Enzyme phosphatase, alkaline: Sigma. • Other chemicals: analytical. Psilocybin and psilocin are the Opium Act. For possession of these substances, but also for the acts to be performed in order to process and mushrooms to analysis, an opium for these substances (site specific) to be present. For orders of standards psilocybin and psilocin abroad is also a Import license (KvW South is owned by an opium for both substances). 2.1.2 Sample Preparation (sample preparation) The dried mushrooms were sampled using an analytical mill ground and in air-tight preserved specimen jars. Fresh mushrooms were sampled immediately after receiving cut into small pieces and a Petri dish for at least 5 days to air dry. Simultaneously, the moisture loss (in%) of these mushrooms determined. In Table 2 (8.2) is the percentage of dry residue (Dry residue =% 100% - moisture%) of the various dried mushroom samples (specimens) displayed. Then the dried mushrooms with an analysis mill ground. Some samples were thus not homogeneous samples; these samples were then ground in a ball mill. The specimen was sealed preserved specimen jars. In the above prepared sample was determined by dry matter content of approximately 100 mg This specimen 150 minutes to dry in an oven with no fan at 102-105 ° C. With this level, the percentage of psilocybin and psilocin are calculated on the dry and to the levels of both substances in fresh and dried mushrooms are compared be (8.2: 1 and Table 2). The mushrooms were not further characterized, the species name was marked on the label used. 2.1.3 Standard and sample solutions Standard psilocybin: Weighing just prior to analysis 1.0 -1.5 mg psilocybin at 0.01mg accurately in a 10 ml flask. Dissolve and make up to 10 ml. Prepared from this solution other desired concentrations for the selection of the calibration line (uitverdunnen). Standard psilocin: Prepared shortly before the analysis using the standard psilocybin the enzyme alkaline phosphatase psilocin the standard. Prepared from this solution other desired concentrations for the selection of the calibration line (uitverdunnen). Sample Solutions: Weigh 250 mg of dried sample in a flask of 25 ml. Add 20 ml of methanol to and place the flask during 120 minutes in an ultrasonic bath. Make sure the temperature in the ultrasonic bath is not above 40 ° C rises. Check the temperature in the bath and add ice increases as the temperature is higher than (>) 40 ° C border. Dilute with methanol to 25 ml mix. Centrifuge (about 16000x g) a portion of the extract and transfer the clear extract two vials (see 2.1.4). 2.1.4. LC The use of reversed phase chromatography is very suitable for biological materials separate and analyze. The LC separation was performed on a Merck RP-60 Lichrospher select B (250 x 4 mm, 5 μm) column with a 4-4 LiChroCART Merck, LiChrospher 60 RP-select B (5 μm) guard column. The components were detected at 266 nm with a UV detector. ________________________________ Page 7 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 7 of 17 Initially it was the intention of both components by gradient elution In an analysis run to determine. The above LC conditions were used: Solvent A: 20 mM KH 2 PO 4 and solvent B: acetonitrile, flow rate constant size1 ml / min., gradient: 5% B for 8 minutes, then rising to 20% B in 20 minutes and then for 10 minutes 20% B (injection volume 5 column oven temperature sterile glycerol 30 ° C). Both substances were well separated. Conditioning and stabilizing the column after the gradient run, however, lasted very long. As a result, the retention times of psilocybin and psilocin after 5 to 10 injections were more or less constant. Based on this experience we decided to separate the two substances by means of isocratic analysis to determine. The psilocybin was using the mobile phase 4% acetonitrile in 20 mM potassium phosphate solution (V / v) and psilocin using the mobile phase 12% acetonitrile in 20 mM potassium phosphate solution (V / v) was determined. The other parameters such as wavelength, injection volume and column temperature remained the same as the gradient. The analysis in this report using these custom analysis obtained. 2.1.5 Robustness Study Both assessments were on a robust examination. As a possible critical factors were considered: • The vibration time of sample extracts in an ultrasonic bath (120 minutes). For both psilocybin and psilocin analysis for the "+" level of vibration time 135 minutes and the '-' chosen level 115 minutes. • The temperature in the ultrasonic bath (up to 40 ° C). For the "+" level was up to 45 ° C and the '-' level up to 35 ° C is selected. When the experiment was checked whether these temperatures during the extraction in ultrasonic bath were achieved, but also has ensured that indicated limits were not exceeded. This factor was critical for both analysis investigated. • The content of acetonitrile in the eluent (psilocybin analysis: 4% acetonitrile in 20 mM buffer solution (v / v); psilocin analysis: 12% acetonitrile in a 20 mM buffer solution (v / v). For the "+" level was both analyzes the content of acetonitrile in the buffer 1% higher and "-" level 1% lower than the declared value chosen. 2.1.6 Validation Study The methods for psilocybin and psilocin were subjected to such validatestudie described in SOP-KAM05 WV102, version 2 of the KvW "Validation of (physico-) chemical and (Physico-) mechanical methods ". The specificity and selectivity of the method were discussed in some detail. Also, the detection limits (3 * Noise), the detection limits (6 * Noise), the correlation coefficients, the recovery and the standard deviations of the two analysis determined. Because the standards were difficult to obtain, an alternative way to apply recovery and standard deviation of each analysis to determine: Extracts of sample survey of mushrooms containing high levels of psilocybin were collected, and mushroom extracts containing high levels of psilocin. These solutions were psilocybin or psilocin content of the quantitatively determined in duplicate. To blank samples were 3 different amounts of these solutions added in duplicate and analyzed. The added amounts of psilocybin / psilocin (known concentration) and analyzed the levels of psilocybin / psilocin could recovery programs at the levels 3 computing (possibly after correction for the blank). 2.2 Monitoring Shrooms In 2001 the project "Maintaining and monitoring the smartshop branch" out. A part of this project was a smart shop in the surveillance industry and in the context of this activity was the sampling and analysis of 40 psilocybin and psilocin or ________________________________ Page 8 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 8 of 17 samples (mushrooms) are planned. The products were two periods of three months (1 e period: March-April-May and 2 e September-October-November) by specially trained inspectors sampled (20 samples per planning period). The levels psilocybin and psilocin in these samples were determined with the developed method. An inspection by an inspector at a mushroom farm showed that the mushrooms and mushrooms flights (growth stages) can be picked. On this farm were 4 types of mushrooms (Types) mushrooms grown, namely the Psilocybe cubensis mexican, the Psilocybe cubensis thai, the Panaeolus cyanescens and Psilocybe Tampanensis. The first three grades were 3 Flights picked, while Psilocybe Tampanensis was harvested at once. With the willing cooperation of the grower could harvest samples immediately after each taken from these types of mushrooms. Because these samples immediately after entering the laboratory to work, could such a reliable picture of the levels and psilocybin psilocin in the fresh mushrooms in various stages of growth are obtained. 3 RESULTS 3.1 Methodology - LC separations Psilocybin and psilocin were determined by separate LC analysis. Both provisions separations were good with the other components in the observed matrix (see chromatograms 1 and 2, see Annex 8.1). Retention times of psilocybin / psilocin were under these circumstances, consistently good. - Robustness Study The investigated factors (vibration time, temperature and% acetonitrile in the mobile phase) were both determination of psilocybin and psilocin in determining (both with 2 different Sample extracts performed) within the margins are not critical. - Selectivity and Specificity The methods for psilocybin and psilocin are only performed in mushrooms and directly related products (eg mushroom tea). Matrix of issues (eg caused by fats and carbohydrates) is thus no question. Including interference effects, which could potentially arise with related components the extract, can be excluded. The only indole alkaloids in solution both to investigate materials and given the difference in polarity of the two compounds, no interference effects to be expected. - Detection and the detection limits The detection limit (3 * Noise) and the limit (6 * Noise) for determining psilocybin in magic mushrooms, respectively 0.005% and 0.009% (w / w). For psilocinebepaling mushrooms in these parameters 0.002% and 0.003% (w / w). - Correlation coefficients The psilocybin and psilocin were over the monitoring analysis and robustness and valdatieonderzoek respectively 7 and 5 once. The calculated calibration lines were linear. The correlation coefficients of psilocybin calibration lines were between 0.9997 and 1.0000. In the calibration lines of psilocin determination were 0.9994 and 0.9999 between the values found. The range of both psilocybin and psilocin in the analysis ranged 0.001 - 0.1 mg per ml of psychoactive (5 points). - Recovery and relative standard deviations The recovery of the provision for the working psilocybin (j w ) Is 93.4% with a relative standard deviation (RSD w ) Of 0.7%. When psilocin provision for recovery area (j w ) 95.8% and relative standard deviation (RSD w ) 1.5%. 3.2 Monitoring Project In total for the monitoring project in 11 smartshops 19 ________________________________ Page 9 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 9 of 17 dried mushroom samples (whether or not ground) and 3 mushrooms containing products sampled. The vast majority of these (19 dried mushrooms and mushroom-containing product 1) was the 1 e period sampled. Besides the sampling of dried mushrooms, fresh mushrooms were also sampled. In the 2 e sampling period, a total of 7 samples taken from 4 fresh mushrooms several smart shops. It was remarkable, all the smart shops sampled fresh mushrooms also the mushroom grower came. This magic mushroom grower were also sent 22 samples of fresh mushrooms taken. All results of the dried mushrooms are shown in Table 1, the results of the fresh mushrooms in Table 2 (8.2). Samples 37085286 and 37099821 are in the table twice. The reason is that Both samples consisted of 2 cups, whose mushroom size, color and consistency differed much from that reprocessing as separate sample was desired. In tables and charts (8.2 and 8.3) are listed codes, which the research laboratory have been applied. These codes correspond to ISI numbers. 4. DISCUSSION 4.1 Methodology Extracting the hallucinogenic substances from the crushed sample with methanol an ultrasonic bath is simple to perform and reproducible. Using both isocratic analysis psilocybin and psilocin were well separated from the other matrix components in the mushroom. Found with recovery programs (determined using the alternative validation procedure § 2.1.6) of 93.4% to 95.8% psilocybin and psilocin is for concluded that both substances properly extracted from the matrix can be mushrooms. The limit of psilocybin is 0.009% w / w and psilocin 0.005% w / w. Muss Hoff [7.7] found a similar method with a limit of quantification of 0.003% 0.01% psilocybin and psilocin on. Both components were in his research, however, an analytical run by applying gradient elution determined. 4.2 Monitoring Project The 2 e sampling period, in contrast to the 1 e period, a joint action by the KvW with the police, all dried samples (Opium Act violation) were the Courts seized. This has the 2 e found significantly less samples (only fresh mushrooms and two tea samples) for research received. The overview of the dried mushrooms in the 1 e sampling period shows that mushrooms Panaeolus cyanescens type of the highest rates of hallucinogenic substances. For total psilocin in dry matter (the best measure for the hallucinogenic effect of mushrooms to be measured) values were between 1.0 and 1.6% were found. Striking was the changing contribution of psilocybin (0.2-0.9%) and psilocin (0.4-1.3%) to total fatty psilocin. Other researchers [7.2 and 7.7] found similar results for psilocybin and psilocin in the mushroom species. Most dried mushrooms (7.9) were called Psilocybe cubensis mexican type, the levels found ranged between 0.7 and 1.2% psilocybin and psilocin in between 0 and 0.3% (dry matter). These values were at the same level as the results by Muss Hoff [7.7] for this type of mushroom found. Total psilocin (On dry matter) of this type of mushroom is between 0.5 and 0.9%. For the sample Psilocybe semilanceata (bald head) were found identical values as for Psilocybe cubensis mexican. In the sample and a azurens Psilocybe cubensis Thai Psilocybe of values were between the 0.5 and 0.9% for total psilocin (on dry weight) measured. The participation in these psilocin samples is considerably higher than the Psilocybe cubensis mexican. The remaining samples, including the mushroom tea samples contain only minor quantities of hallucinogenic substance (<0.05%). ________________________________ Page 10 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 10 of 17 The psilocybin and psilocin levels in fresh samples of Panaeolus cyanescens mushroom grower showed a varied picture. Some samples were very high psilocybin rates (up to 3% dry matter) combined with relatively low rates to psilocin (up 0.5%) were measured. In contrast, other samples were high percentages psilocin (up 1.8% on dry substance) were found and the very low rates of psilocybin (<0.1%). The form in which hallucinogenic substances are present, is random, at one time the hallucinogenic substance psilocybin largely in the form of existing, sometimes almost entirely in the form of psilocin. One explanation for these large variations could not be given. The total psilocin (on dry matter) varies widely, from 0.8% to 2.5%. This rates of hallucinogenic substances, in whatever form, are some freshly picked Panaeolus cyanescensmonsters is significantly higher than other investigators [7.2 and 7.8] published in the past (and also higher than in the investigated variant of dried This type of mushroom). The applied drying and harvest time are the possible causes lower rates of psychoactive substances in the dried form. When comparing the Panaeolus cyanescensmonsters, originating from different flights (growth stages, and the mushrooms picked) were derived, no association between the levels psilocybin / psilocin and the flight from which the samples were obtained (Chart 3). The psilocybin, psilocin, and the total psilocinegehalte (0.5-0.9%) in fresh Psilocybe mexican cubensis samples were at the same level as in the dried samples from smartshops. The Psilocybe cubensis thai appears not only in appearance but also in the analysis concerns, much like the mexican cubensis Psilocybe. The detected levels of psychoactive substances In this type of mushrooms were only marginally lower (total psilocin: 0.5 -0.7%). The levels of psychoactive substance in the fresh Psilocybe cubensis thai, taken in a smartshop (code 41 and 43), however, from the same mushroom grower, were particularly low. This indicates that psilocybin (psilocin that breaks through) and psilocin in the fresh mushrooms much less stable than in the dried mushrooms. This is also consistent with experience gained with demo samples. Two new demo samples (1 Psilocybe cubensis Thai Psilocybe cubensis mexican and 1), both sampled the mushroom grower (For an instruction session), were dried and analyzed after 4 weeks. In both samples could be demonstrated only traces hallucinogenic substance. There were also the Psilocybe cubensis mexican and thai no relationship was observed between both the percentages psilocybin, psilocin and total psilocin and the flight, which the samples were taken (8.3: Chart 3). Total psilocin (the solids) in Psilocybe Tampanensis samples was approximately 0.3%. Hallucinogenic substances (and psilocybin) mushrooms in this species much more stable than the other investigated fresh mushrooms. The smart shops in the sampled Psilocybe Tampanensis samples contain fact, despite the dry residue by wastage (during storage in smartshop) was increased to 60-65%, almost as many psychoactive substance and fresh harvested samples. A demo-sample, which, like the above samples even after 4 weeks were dried and analyzed, had a total of nearly 0.3% psilocin. 5. CONCLUSION The indole alkaloids psilocybin and psilocin can by using methanol extraction with the reversed phase LC technique quickly and accurately determined mushrooms. The levels of psychoactive substance same type layers in the dried mushrooms and in the fresh form roughly the same level and agreed to in the literature values. Exception is the occasional but very high levels of psilocybin and psilocin in the fresh Panaeolus cyanescens. There were no relationships observed between the psilocybin, psilocin and total psilocinegehalte and the flight from which the mushrooms came. The investigation also revealed that the psychoactive substances psilocybin and psilocin fresh mushrooms far less stable than in the dried form. ________________________________ Page 11 SAZD/01/30/21 - psilocybin and psilocin in magic mushrooms SAZD/01/30/21 Page 11 of 17 6. THANKS The authors of this report to Dr. WG van der Sluis of the Faculty of Pharmacy, Department Pharmacognosy, University of Utrecht to thank for his advice, free availability of standard equipment and the procedure for psilocybin psilocin from this standard can be prepared. 7. -------------------- Cloneufc's easy Penis Envy spore tek! Cloneufc's easy Cloning tek Cloneufc's 360 degree LC Cloneufc's No foil jar tek! Cloneufc's Breaking up spores
| |||||||
|
Ascension Energy & Alien UFOs Registered: 05/12/07 Posts: 71,179 Loc: The Inexpressible... Last seen: 3 hours, 18 minutes |
| ||||||
|
Wow Cloneufc, that is an amazing article find. Good job!
![]() Those Dutch always get the cool illegal experiments. I'm going to read the whole thing later, but the second graph on the bottom, comparing the active concentrations of 3 types of psychoactive species, is incredible! Its exactly what I want to see done for *just* for Ps. Cubenis compared to Penis Envy...that would be the ultimate experiment result. Also, how's that Gas/Liquid Spectrometry machine doing? Are you still considering buying one? Excellent On-Topic Article! ~ LogicaL Chaos ~
| |||||||
|
Master Exploder! Registered: 11/15/07 Posts: 1,237 Loc: Las Vegas Last seen: 1 year, 5 months |
| ||||||
|
I have the money to buy the machine now. Im holding off buying it for two reasons. First I need to buy new stuff,house, cars, toys ect. Second I want to make sure I buy the correct equipment, I dont want to waste money buying the wrong machine. Before I buy it I need to make sure 100% that it will do what I need it to do.
Wow the Pan Cyan hawaii sample was way stronger than the Azure sample. That surprises me. The P. tampanensis was weak as hell. Am I reading this correctly? Is it this easy to prepare the sample? Why do you need to transfer the sample to two vial instead of one? Weigh 250 mg of dried sample in a flask of 25 ml. Add 20 ml of methanol to and place the flask during 120 minutes in an ultrasonic bath. Make sure the temperature in the ultrasonic bath is not above 104 ° F rises. Check the temperature in the bath and add ice increases as the temperature is higher than (>) 104 ° F border. Dilute with methanol to 25 ml mix. Centrifuge (about 16000x g) a portion of the extract and transfer the clear extract two vials. -------------------- Cloneufc's easy Penis Envy spore tek! Cloneufc's easy Cloning tek Cloneufc's 360 degree LC Cloneufc's No foil jar tek! Cloneufc's Breaking up spores Edited by Cloneufc (04/08/10 11:48 AM)
| |||||||
|
liberator Registered: 08/06/08 Posts: 4,204 Last seen: 4 years, 6 months |
| ||||||
|
So who's game for a revival of this? Shea25 posted this recently, looks pretty legit:
http://www.thenook.org/nooki/ind It's an ab extraction, and the end result gets crystals. The writer of the tek says that it is 70% efficient, which is pretty damn efficient if you ask me. I intend to try this on the OI and OR I just made lc's of once I have fruits from them, and then later down the line try it on PE whenever that grow materializes, which probably won't be for at least a few months. Anyone have any fruit on hand they'd like to try this on? Everything is available at your local grocery store or hardware store. So whadya think, are we back in business? Anyone wanna try this out?
| |||||||
| |||||||
|
| Similar Threads | Poster | Views | Replies | Last post | ||
![]() |
Melatonin Super Charge Potency Project. | 855 | 0 | 09/02/05 10:29 PM by Ogla | ||
![]() |
Anyone ever successfully extracted psilocin/psilocybin? | 2,185 | 9 | 01/16/06 02:19 AM by mogur | ||
![]() |
Notes from a failed psilocin extraction (update: 2nd attempt) ( |
8,668 | 21 | 06/30/05 08:43 AM by triptamine | ||
![]() |
Will low heat affect potency? Answer=No ( |
25,131 | 34 | 08/21/18 02:51 PM by bodhisatta | ||
![]() |
VERY low potency from hpoo shrooms why?(NOT B/C OF DEHYDRATOR) ( |
9,480 | 45 | 02/26/06 06:29 PM by HUBSonDUBS | ||
![]() |
Potency Differences Between Caps and Stems... ( |
104,981 | 35 | 03/10/20 01:28 AM by bodhisatta | ||
![]() |
Re: psilocybin/psilocin extraction tek ( |
9,683 | 36 | 06/28/00 03:17 PM by Anonymous | ||
![]() |
potency and temperature | 5,670 | 15 | 04/02/03 06:33 PM by Smiley_Riley |
| Extra information | ||
| You cannot start new topics / You cannot reply to topics HTML is disabled / BBCode is enabled Moderator: Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, LogicaL Chaos, 13shrooms, hamloaf, cronicr, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a 29,628 topic views. 14 members, 92 guests and 28 web crawlers are browsing this forum. [ Show Images Only | Sort by Score | Print Topic ] | ||



