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Fanaticus (Fanaticus)
Posted on Friday, October 05, 2001 - 06:14 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Check this out - there is some really pertinent info here - lets discuss this!

from the FORENSIC SCIENCE INTERNATIONAL journal.
113 (2000) 389-395

Hallucinogenic mushrooms on the German Market-
simple instructions for examination and identification.

it says that "The cultivation or possession of whole
Psilocybe mushrooms and its spores are
restricted by German law since 1998"

They analyzed four magic shrooms available over the counter
in Holland. Cubensis - Lib cap - Pan cyan and Tampanensis

they analyzed
18 specimens of Psilocybe Cubensis
9 specimens of Lib cap
6 specimens of pan cyan
4 specimens of tampanensis

------------------------------------------------
Psilocybe Cubensis

psilocybin (%) psilocin (%)
none ------- .14
none ---------- .05
none ------- .10
none (detected)--- ---- .10
none --- ---- .11
.01 --- ---- .05
.02 --- ---- .09
.17 --- ---- .09
.31 ------- .23
.50 ------- .12
.87 ------- .04
.98 ------- .03
1.07 ------- .01

------------------------------------
PSILOCYBE SEMILANCEATA

psilocybin psilocin
.01 ---- --- .48
.16 ---- --- .13
.25 ---- --- .08
.27 ---- --- .24
.30 ---- --- .03
.42 ---- --- .04
.51 ---- --- .12
.72 ---- --- .01
.91 ---- --- .90

--------------------------------
PAN CYAN

psilocybin psilocin

.02 ---------- .56
.44 ------- .14
.47 ------- .22
.51 ------- .64
.54 ------- .09
1.15 ------- .90

--------------------------------
PSILOCYBE TAMPANENSIS

psilocybin psilocin

none ------- .02
.01 ------- .03
.03 ------- .03
.19 ------- .01
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monkeyod (Monkeyod)
Posted on Friday, October 05, 2001 - 07:31 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Most of those numbers seem way low. Might be from the way they dried them? Nice to see your not awol PF...your site goes down for a day or two and some people start thinking bad things.
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Lichen (Lichen)
Posted on Friday, October 05, 2001 - 09:20 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Well, I have to agree...some of the cubies they tested showed no psilocybin. However, some showed strong. Why would that be?

Also, it's interesting to note that some of the cubies were as strong in psilocybin as almost any other shroom they tested. From this, it would seem cubensis isn't so weak as we were led to believe.
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greenthumb (Greenthumb)
Posted on Saturday, October 06, 2001 - 09:45 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

erowid.com has a list/chart of mushroom potency that when compared to this list seems to cast even more doubt in my mind about the knowledge of the potency of mushrooms. Are the inconsistencies a matter type, cultivation, processing, age, or all of the above? I think all of these things could affect any data gathered from such small test groups. The data gathering used to support the Erowid potency chart is unknown to me. I think that large test groups with good control would be tough because of the tricky / picky grow teks required to produce certain varieties. This said, I think one can only generalize when saying type A is more potent than typeB. TypeA grown on dung may be less potent than type A on a PF cake.
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jim brown (Shrhobbyist)
Posted on Sunday, October 07, 2001 - 09:57 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Did they happen to mention how these levels were found? Did they note any differences among the mushrooms of the same type (race, weight, age, place found, way they were grown...)?
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Lichen (Lichen)
Posted on Sunday, October 07, 2001 - 10:40 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

this is a very astute observation; fanaticus' opinion is that psilocybin shrooms grown on brown rice are 'an entheogen of the highest order'. But what if grown on some other substrate?

My friends who have shared my shrooms say they cannot tell the difference between grass- and rice- and dung-grown shrooms. Neither can I. But what would a bio-assay show?
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Sillycybin (Sillycybin)
Posted on Monday, October 08, 2001 - 04:07 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

I think I'll probably just be saying what has already been said, but I read the article and I don't think we can draw any meaningful conclusions about relative potency from this article. I think the purpose of the article is as its title anounces--how to examine and identify 'magic' mushrooms.

Some other pertinent information when drawing conclusions:

-They also said that when extracting the psychoactive ingredients for toxicological examination, the samples were 'ultrasonicated' for 120min with a max temp of 50C, which is 122F.

So the samples were heated, destroying some of the psychoactive ingredients. So we can't ge acutal quantities of psilocin/psilocybin contained in the samples.

Even so, if they were all treated in the same manner, one would suspect that one could draw conclusions about relative potency, since ratios might stay the same. However:

-They said that the samples were "confiscated by local authorities".

There is no control on this experiment. The samples were not grown by those performing the experiment, with the only difference being the species. It is not known if the samples were all grown in the same manner. It is not known if the samples were all preserved in the same manner. Their purpose was to identify the different samples as psilocin/psilocybin containing and to identify the species.

-There were only 37 samples--the majority being from one species (as PF pointed out)

Personally, I don't think that 37 samples is a large enough sample group to draw any conclusions about potency among species. Especially if there is more data on one species than the others.


Just my two cents
Silly
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Sillycybin (Sillycybin)
Posted on Monday, October 08, 2001 - 05:37 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Copied without permission from:

Forensic Science International

Volume 113, Issues 1-3
SummaryPlus
11 September 2000

Article
Pages 389-395

PII: S0379-0738(00)00211-5
Copyright © 2000 Elsevier Science Ireland Ltd. All rights reserved.

Hallucinogenic mushrooms on the German market ¯¯ simple instructions for examination and identification

F. Musshoff, , a, B. Madeaa and J. Beikeb

a Institute of Legal Medicine, Rheinische Friedrich-Wilhelms-University, Stiftsplatz 12, 53111 Bonn, Germany
b Institute of Legal Medicine, Westfaelische-Wilhelms-University, von-Esmarch Strasse 62, 48149 Muenster, Germany

Available online 6 September 2000.


Abstract

`Magic mushrooms' is the name most commonly given to psychoactive fungi containing the hallucinogenic components psilocybin and psilocin. Material confiscated by local authorities was
examined using morphologic, microscopic, microchemical, and toxicological methods. Psilocybe cubensis was the most popular mushroom in the sample collective, followed by Psilocybe
semilanceata, Panaeolus cyanescens and Psilocybe tampanensis. The alkaloid content was determined with <0.003¯1.15% of psilocybin and 0.01¯0.90% psilocin. Panaeolus cyanescens
was the mushroom with highest levels of psilocybin and psilocin.

Author Keywords: Hallucinogenic mushrooms; Psilocybe; Psilocybin; Psilocin; Morphology; Analytical toxicology


Article Outline

1. Introduction
2. Material and methods
2.1. Chemicals
2.2. Macroscopic examination
2.3. Microscopic examination
2.4. Microchemical examination
2.5. Toxicological analysis
3. Results
4. Discussion
References


1. Introduction

`Magic mushrooms' is the name most commonly given to psychoactive fungi [1]. From hallucinogenic mushrooms Hofmann et al. [2] isolated two hallucinogenic components of the tyramine type,
psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine), the main psychotropic compound, and psilocin (4-hydroxy-N,N-dimethyltryptamine). The metabolism and pharmacokinetic properties of
psilocybin have been investigated [3]. The rapid and extensive cleavage of the phosphoric ester group of psilocybin by alkaline phosphatase and unspecific esterases indicates that psilocybin acts as
a prodrug and that its hydroxy metabolite psilocin represents the true pharmacologically active agent. 4-Hydroxyindole-3-acetic acid and 4-hydroxy-tryptophole were identified as further
metabolites. Today these indole alkaloids have become biochemically important drugs in psychotherapy and psychodiagnostics. Moreover, ancient cults using magic mushrooms are reborn as a
ritual in the drug scene [4, 5 and 6]. Fresh or dried Psilocybe mushrooms and also kits consisting of spores, fertiliser and instructions on how to grow cultures are offered worldwide. German
consumers get their supplies through the Internet or they buy material in Dutch `Smart-Shops'. Some of the material has been confiscated by local authorities. With increasing frequency we are
asked to analyse these materials. We report about established methods and common findings in morphologic, microscopic, microchemical, and toxicological examination of hallucinogenic
mushrooms.

2. Material and methods

2.1. Chemicals

Psilocin and psilocybin were purchased from Alltech (PA, USA). The other chemicals were obtained from Merck (Darmstadt, Germany). Ehrlich's reagent: 5 g 4-dimethyl-aminobenzaldehyde in
50 ml of 90% ethanol to which 50 ml of 38% hydrochloric acid is added. Melzer's reagent: 1.5 g iodine in 100 ml of an aqueous mixture containing 5 g of potassium iodide and 100 g of chloral
hydrate.

2.2. Macroscopic examination

Our material consisted of 37 confiscated samples of dried material. Diagnosis of genus and species of dried hallucinogenic mushrooms is extremely difficult, because the samples often consist
either of broken parts of caps and stems or of tissue ground to granules or powders. For the latter, microscopic examination revealed further results. We did not find fresh material, which might
have been used for a spore print or the bluing reaction for identification purposes.

2.3. Microscopic examination

A 10% ammoniacal solution (v/v) acts as a mild clearing agent and possesses a contrasting refractive index. It also acts as a swelling agent and allows the dried tissues to return to their previous
state, when the agaric was fresh. An extensive microscopic examination is described by Watling [7] who studied three parts of a mushroom, the gill, the scalp of the cap and a longitudinal section
through the gill. Spores free of tissue can be obtained from the sample simply by washing the gills in the 10% ammonia.

2.4. Microchemical examination

Melzer's reagent when placed on tissues or spores may turn them blue or bluish-black, purple-brown, or golden to yellowish. The colour change is characteristic for the fungi under study. If they
become blue, bluish-grey or blue-black they are amyloid, and if they are unchanged or only become slightly yellowish they are termed inamyloid or non-amyloid. Mushrooms of the genus
Psilocybe are always inamyloid.

Treatment of dried mushroom samples with Ehrlich's reagent for 30 min may results in a purple or greyish-purple colour reaction in the presence of primary aromatic amines, i.e. the hallucinogenic
alkaloids of the Psilocybe genus.

2.5. Toxicological analysis

For the extraction of psilocybin and psilocin from mushrooms 100 mg of dried and pulverized plant material are added to 9 ml methanol. This mixture is ultrasonicated for 120 min (maximum
temperature of the mixture 50°C). Then the volume is brought to 10.0 ml with methanol; the mixture is centrifuged and the supernatant is used for the HPLC determination. A Hewlett-Packard
Series II 1090 liquid chromatograph equipped with a Hewlett-Packard Series 1050 variable wavelength detector (266 nm) was used. Chromatographic separation was performed with a
LiChrospher 60 RP-select B (250?4 mm, 5 m) analytical column (Merck) and gradient elution was carried out at a constant flow-rate of 1 ml/min. Solvent A consisted of 20 mM KH2PO4 and
solvent B of acetonitrile. Initial conditions were 5% B for 2.5 min increasing to 25% B at 15 min. For the calibration aqueous standard solutions of psilocybin and psilocin in methanol in the
concentration range 0, 10, 50 and 100 g/ml were used.

3. Results

Using Melzer's reagent all samples tested were inamyloid. Further microchemical investigation of our material resulted in most cases in a positive reaction using Ehrlich's reagent. By toxicologic
analysis it was confirmed that all positive samples contained psilocybin or psilocin.

Photographs, descriptions of morphologic characteristics, and toxicologic analysis are summarized in Fig. 1, Fig. 2 and Fig. 3 and Table 1. Microscopic examination revealed in most cases fungal
tissues consisting of parallel hyphae in which adhering cells were connected to clamp connections. The spores we recognized measured 3¯10 m by 12¯20 m and were elliptic or lemon-shaped,
brown to black in color with markedly thick walls, smooth surfaces and an easily visible germ-pore (Fig. 4).


fig1 (31K)

Fig. 1. Panaeolus cyanescens.


fig2 (21K)

Fig. 2. Psilocybe semilanceata.


fig3 (31K)

Fig. 3. Psilocybe cubensis.


tab1
Table 1. Morphologic characteristics of hallucinogenic mushrooms
(37K)


fig4 (43K)

Fig. 4. Spores of Psilocybe cubensis (magnification?1250).


tab2
fig5
The results of the toxicological analysis are summarised in Table 2. The calibration curves are linear in the range 0¯100 g psilocybin and psilocin/ml (limit of quantification, 0.01%; limit of
detection, 0.003%). The simple and rapid extraction procedure results in chromatograms without interfering peaks (Fig. 5).


Table 2. Results of toxicological analysis
(12K)


(4K)

Fig. 5. Chromatogram of an extract from a Psilocybe cubensis mushroom, containing 0.31% psilocybin and 0.23% psilocin.


4. Discussion

Macroscopic examination of dried hallucinogenic mushrooms is extremely difficult, because the samples often consist of broken parts or powders. In extreme instances microscopic examination
can reveal further results. The microscopic evaluation of tissues consisting of parallel hyphae in which adhering cells are connected to clamp connections are a definite proof that the material is of
fungal origin. In addition spores are of interest for identification purposes. All spores of Psilocybe mushrooms are of similar shape, brownish, and smooth. The validity of microchemical reactions
is limited. The reaction with Melzer's reagent may serve only as an exclusion diagnosis. A positive reaction with Ehrlich's reagent depends on the alkaloid content, which can be decreased in stored
material.

Morphologic examination revealed Psilocybe cubensis as the most popular mushroom in our sample collective, followed by Psilocybe semilanceata, Panaeolus cyanescens, and Psilocybe
tampanensis. The alkaloid content was determined with <0.003¯1.15% of psilocybin and 0.01¯0.90% psilocin. Panaeolus cyanescens was the mushroom with highest levels of psilocybin and
psilocin.

Psychoactive mushrooms are either eaten fresh, which supposedly produces the most powerful and intense visual experience, or dried and consumed at a later date [8]. Some users freeze
mushrooms for later use, while others put them in capsules for resale. Mixing the psychoactive fungi in milk shakes or tea or cooking the mushrooms in a soup, stew or omelette has also been
reported. Boiling the mushrooms in water to remove the active, hot water-soluble ingredients and use of the water to prepare foods such as rice or others is another ingestion form. Chocolate and
honey are also employed by recreational users for use with mushrooms and is a form for transport and export. Mushrooms in honey preserve their potency much better than those that are dried or
frozen, especially if placed in honey immediately after having been picked.

Symptoms produced by eating psychoactive mushrooms begin to occur within 20¯40 min after ingestion or from 5 to 10 min when prepared in the form of soup or tea. Symptoms persist for up to
4¯8 h after ingestion. The hallucinogenic effects are similar to those observed after LSD intake. However, the dose of psilocybin/psilocin required for the same effect is approximately 200 times
higher. The usual dose of psilocin required to induce psychedelic effects ranges from 8 to 10 mg [9].

In Germany psilocybin and psilocin are classified substances (§1, Abs.1, Anl.1 BtmG). The cultivation or possession of whole Psilocybe mushrooms and its spores are restricted by German law
since 1998 (10. BtmÄndV).


References

1. R.G. Wasson, Seeking the magic mushroom. Life 42 (1957), pp. 100¯120.

2. A. Hofmann, A. Frey, H. Ott, T. Petrzilka and F. Troxler, Konstitutionsaufklärung und Synthese von Psilocybin. Experienta 15 (1958), pp. 397¯399.

3. F. Hasler, D. Bourquin, R. Brenneisen, T. Bär and F.X. Vollenweider, Determination of psilocin and 4-hydroxyindole-3-acetic acid in plasma by HPLC¯ECD and pharmacokinetic profiles of
oral and intravenous psilocybin in man. Pharm. Acta Helv. 72 (1997), pp. 175¯184. Abstract | Journal Format-PDF (756 K)

4. A. Gottlieb. Psilocybin Production, Ronin, Berkeley, CA (1997).

5. G.P. Menser. Hallucinogenic and Poisonous Mushroom ¯¯ Field Guide, Ronin, Berkeley, CA (1997).

6. P. Stamets. Psilocybin Mushrooms of the World, Ten Speed Press, Berkeley, CA (1996).

7. R. Watling, Hallucinogenic mushrooms. J. Forensic Sci. Soc. 23 (1983), pp. 53¯66. EMBASE

8. J.W. Allen, M.D. Merlin and K.L.R. Jansen, An ethnomycological review of psychoactive agarics in Australia and New Zealand. J. Psychoactive Drugs 23 (1991), pp. 39¯69. MEDLINE
EMBASE

9. T. Geschwinde. Rauschdrogen: Marktformen und Wirkungsweisen (3. Aufl), Springer Verlag, Heidelberg, Berlin, New York (1996).
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Black Star (Mr_Bug)
Posted on Monday, October 08, 2001 - 05:47 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Would someone be so kind as to decode the above posting?
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Sillycybin (Sillycybin)
Posted on Monday, October 08, 2001 - 06:39 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

If we want to know the potency of shrooms, can't we theoretically do our own experiments without fancy mass spectrometers and the like?

If we weigh our sample beforehand, then do an extraction technique with some solvent, allow the solvent to evaporate...wouldn't you eventually get crystals of psilocin/psilocybin if you evaporate all the solvent? Then weigh that, and you have a % by weight...then find the % by weight for several species, and you can do a comparison? If you're looking for potency, wouldn't that be the way to compare?

Or does psilocin/psilocybin crystallize? I don't really know enough about the subject matter...

Sillycybin
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Sillycybin (Sillycybin)
Posted on Monday, October 08, 2001 - 06:39 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Don't know why it double posted--sorry

Sillycybin