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Mycotopia Web Archive ´┐Ż Archive ´┐Ż Drug Dosage & Dosing: Recipies & Extracts. ´┐Ż Potency variations in cubensis ´┐Ż Variations of Psilocybin and Psilocin Levels ´┐Ż Previous Next ´┐Ż

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Nan (Nanook)
Posted on Tuesday, October 23, 2001 - 07:20 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Here is the original Bigwood and Beug paper on potency variations.

I only omited the three photographs which just show shrooms in a jar and freezed dried shrooms in a plastic baggie, neith of which photograph was good enough to reporduce here.

mjshroomer
Variations of Psilocybin and Psilocin Levels with Repeated Flushes (Harvests) of Mature Sporocarps of Psilocybe Cubensis (Earle) Singer.

By
Jeremy Bigwood and Michael W. Beug
(Evergreen State College, Olympia, Washington.

Summary

Analysis of Psilocybe cubensis (Earle) Singer) grown in controlled culture showed that the level of psilocin was generally zero in the first (or sometimes even the second) fruiting of the mushroom from a given culture and that the level reached a maximum by the fourth flush. The level of psilocybin, which was nearly always at least twice the level of psilocin, showed no upward or downward trend as fruiting progressed, but was variable over a factor of four. Samples obtained from outside sources had psilocybin levels varying by over a factor of ten form one collection to the next.

Introduction

When undertaking quantitative analysis of psilocybin and psilocin levels in the Pacific Northwest species, we generally found large variations from one collection to another even within one species and even when all collections were made from a single location (Beug and Bigwood, 1982). In investigating biosynthetic pathways in the formation of psilocin and psilocybin in Psilocybe cubensis (Earle) Singer, we also observed variations in psilocybin and psilocin levels from one fruiting to the next (Chilton, 1979). WE therefore stet out to grow a selected Amazonian strain of Psilocybe cubensis (Earle) Singer in carefully controlled cultures and study the variations of psilocybin and psilocin levels with time. We also report here on the observed variation of psilocybin and psilocin levels with repeatted flushes from a single culture and the variation observed in other strains.

Experimental

The strain of Psilocybe cubensis cultivated in this study originated from a spore print taken in the Amazon basin near Pucalpa, Peru (Repke et al., 1977). Mycelium obtained from the spore print was kept as a stock culture on various agars. Since only one flush (fruiting) could be obtained from agar plates, we used a rye-grain medium, described initially in San Antonio (1971), refined by Oss and Oeric (1976), and adapted to "miniculture" by us. A wide-mouth half-pint jar (~250 ml) was charge with 10 g of rye grain and 15 ml of water and autoclaved. It was then inoculated under sterile conditions with a mycelium culture on agar. Every four days for a period of 28 days, weight per miniculture. Each flush was harvested as soon as the sporocarps were mature. The mushrooms ere the jars were shaken to distribute the growing mycelium evenly on the grain. In 28 days, the mycelium had covered the grain and the jars were then opened and the grain was cased (covered with a layer about 2 cm deep) with 2 parts peat : 1 part calcium carbonate : 2 parts perlite and/or vermiculite. The mushrooms were "watered" once every two days with 1 ml of sterile water via syringe. The first flyush (fruiting) occurred four to five weeks after inoculation (about two weeks after casing). The minicultures continued to produce mushrooms for at least 20 weeks provided they remained uncontaminated. They yielded an average of 2.7 g dry weight per miniculture. Each flush was harvested as soon as the sprocarps were mature. The mushrooms were immediately freeze-dried, sealed in plastic and stored at´┐Ż5 degrees Celsius until analysis. Voucher specimens were prepared for deposit in the University of Washington Herbarium (WTU).

The extraction procedure and analysis was described in the previous paper. The reversed-phase high performance liquid chromatograms were quantified wioth a Hewlett-Packard 3380! Reporting integrator-plotter and calibrated against standards from the National Institute on Drug Abuse. We found a linear relationship (plus/minus 10% repeatability) between concentration and peak area from 0.2 to 3 micrograms total psilocybine or psilocin. The detection limit was about 0.01 micrograms psilocybin or psilocin. The HPLC results were qualitatively confirmed by TLC using butanol-acetic acid-water (12:3:5).

Results

We found that the levels of psilocybin varied somewhat unpredictably from one flush to the next, but generally were much the same on the last flush as they were on the first flush (Table 1). Psilocin, on the other hand, generally was absent in the first one or two flushes, each maximixed by the fourth flush, and then appeared to start to decline (Table 1). Unfortunately, we could generally not follow the decline appreciably since five flushes is normally the maximum we can get before the mycelium stops fruiting. (With miniculture 1, we obtained a sixth flush but the fifth flush was totally consumed in another experiment and is not reported here.)

In two other strains grown by other sources, we also observed nearly complete absence of psilocin in the first flush . In These, we analyzed the caps and stems separately and found that the caps generally contained twice as much psilocybin as the stems, but that the small amount of psilocin present was entirely in the stems (Table 2). In contrast, our Amazon strai hasd a trace of psiloin in the cap but not in the stem. The cap and stem contained equal amounts of psilocybin.

Finally, we analyzed five street samples of Psilocybe cubensis for which we did not know the flush number or the precise growing conditions (Table 3). We found highly variable levels of psilocybin and low levels of psilocin.

TABLE 1
The dry weight variation of psilocybin and psilocin levels in Psilocybe cubensis as a function of flush number (quantified by HPLC.
Miniculture No. 1
Psilocybein´┐ŻPsilocin
(mg/g) (mg/g)
1 8.3---------------0.5
2 6.5---------------1.5
3 13.3---------------1.0
4 4.8---------------2.6
5 --/--------------/--
6 6.8---------------0.5

Miniculture No. 2
Psilocybin´┐ŻPsilocin
(mg/g) (mg/g)
1 5.1---------------0
2 7.3---------------0
3 4.7---------------1.7
4 3.7---------------2.9
5 5.2---------------2.2
6 --/--------------/--

Miniculture No. 3
Psilocybin´┐ŻPsilocin
(mg/g) (mg/g)
1 7.6---------------0
2 6.2---------------0
3 5.3---------------0.9
4 3.2---------------1.8
5 6.7---------------1.7
6 --/--------------/--

TABLE 2
Distribution of psilocybin and psilocin in the cap versus the stem in three strains of Psilocybe cubensis cultivated on rye-grain substrate
M. R. strain-
First flush
Psilocybin Psilocin
(mg/g) (mg/g)

Caps 9.7---------0
Stems 4.2---------0.35


Equadorian Strain
First flush
Psilocybin Psilocin
(mg/g) (mg/g)

Caps 7.6--------0
Stems 4.7--------0.4


Amazon Strain
First flush
Psilocybin Psilocin
(mg/g) (mg/g)

Caps 5.7--------0.1
Stems 5.7--------0


TABLE 3
Psilocybin and psilocin levels in dried psilocybe cubensis "street samples" (all samlples were from material cultivated on a rye-grain substrate)

Sample Psilocybin Psilocin
No. (mg/g) (mg/g)
1---------------5.6----------------0
2---------------6.2----------------0
3---------------0.7----------------0.3
4---------------0.7----------------0.3
5---------------1.3----------------0.3

Conclusions


We found that the level of psilocybin and psilocin varies over a factor of four among various cultures of Psilocybe cubensis grown under rigidly controlled conditions, while specimens from outside sources varied tenfold. IT is clear that entheogenic (Ruck et al., 1979) and recreational users of this species have no way of predicting the amount of psilocybin and psilocin that they are ingesting with a given dry weight of the mushrooms. It thus seems likely that variations in the subjective experience will not only come from the effects of set and setting but will also stem in very real measure rom large dosage differences.

References

Beug, Michael W. and Jeremy Bigwood. 1982. Psilocybin and psilocin levels in twenty species from seven genera of wild mushrooms in the Pacific Northwest, U.S.A. Journal of Ethnopharmacology vol. 5:271-285.

Chilton, Scott., Bigwood, Jeremy and R. E. Jensen. 1979. Psilocin, Bufotonine and serotonin : Historical and Biosynthetic Observations. Journal of Psychedelic Drugs Vol. 11:61-69.

Oss, O. T. and Oeric, O. N. 1976. Psilocybin Magic Mushroom Growers Guide. And/Or Press, Berkeley, California.

Repke, Carl A. P., Leslie, Dale T., and Gastón Guzmán. 1977. Psilocybe, Conocybe and Panaeolus. Lloydia Vol. 40:566-578.

Ruck,Carl A. P., Bigwood, Jeremy., Staples, Danny., Ott, Jonathan and R. Gordon Wasson. 1979. Entheogens. Journal of Psychedelic Drugs Vol. 11:145-147.

San Antonio, J. P. 1971. A laboratory method to obtain fruit from cased grain spawn of the cultivated mushroom : Agaricus bisporus. Mycologia vol. 63:16-21

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