~ybin in Finnish Psilocybe semilanceata

for C)OH4604: 470.340), 452 (19) (M-~. 424 (9) (452 - COl, 409 (lO) (424 -!lPJ' _68 (10),246 (18), 205 (58), 187 (51),81C? ';5 (100); addition of diazomethane af-~d the corresponding methylester; IH_

'Il. (eDCl), Ò ppm): 0.92,0.95,0.98,1.02,~ and 3.67 (3H each, s), 4.97 and 4.92~ IH. br, s), 4.12 (2H, br, s), 2..:..88(lH,~,J =011. 11,5),2..:..49 (1H, d~d,] - 17, lO,

'40 (IH. ddd , J - 17,7,4),~.-.589 578 546 436 nm

. ~ '" _ (CHCt); c = 0.64)<!tr. +10+11+14+18

S{J./2_Methylbutyryloxy]-preeupatundin-,t((l,l4.epoxide (2): Colourless oil; IR Vc;;,~~13,jlI.l: 3600 (OH), 1765 (y-Iactone), 1730

STRZj2.3277

(C02R); MS: 260.105 (2) (M-RCOzH) (calc.for ClsH1604: 260.105), 242 (6) (260 - HzO),85 (32) (C4H9CO+), 57 (100) (85 - COl; IH_NMR (CDCt), /) ppm): 4.84 (1H, br, d, J = 6,2-H), 5.73 (1H, br, s, 3-H), 2.61 (1H, br, t, J= lO, 5-H), 4.69 (1H, dd, J = lO and 9, 6-H),3.18 (1H, dddd, J = 9,4,3,3, 7-H), 5.51 (IH,ddd, J = 8,8,4, 8-H), 2.91 (1H, ddd, J = 15,9,1.5, 9a-H), 1.97 (1H, dd, J = 15, 9, 9~-H),6.35 (1H, d, J = 3.5, 13-H), 5.54 (lH, d,] = 3,13'-H), 2.77 (1H, d, J = 5, 14-H), 2.71 (IH,dd, J = 5, 1.5, l4'-H), 2.00 (3H, br, s, 15-H),2.30 (1H, tq , J = 7,7, 2'-H), 0.83 (3H, t, J =7, 4'-H), 1.04 (3H, d, J = 7, 5'-H),

589 578 546 436 nm[an40 = (CHCl); c = 0.2)

-32 -32 -36 -60

Psilocybin in FinnishPsilocybe semilanceataJ. Jokiranta I, S. Mustola I, E. Ohenoja ' and M. M. Airaksinen 1.3

\bstract: The use of a hallucinogenic mush-~:\)m.Psilocybe semilanceata, has been occa-sonally reported in Finland, where the spe-:!S iswidely distributed. We have determin-:j, by HPLC. the content of psilocybin and:'\ilocinin P. semilanceata samples collected~ùmdifferent parts of Finland; the psilocybin;~ntent was found to be high (0.62-2.37 %,~,ean1.42 % of dry weight). some samples al-" COntained low concentrations (0.01-.02 % dry weight) of psilocin.

Introduction

The traditional use of psilocybin-:ontaining mushrooms in Centrai Ame-rt\:awas thc focus of intense scientific in-;erest in the 1950s. The psychoactive ef-:,~ctscf psilocvbin and its active metabo-~te, psilocin. ;re similar to those of LSD.lisnow known that psilocybin is presentnotonly in some tropical species, but al-:tn some mushroom species in different

ns of the world (1). and their ingestion

~----------------loe~art~ent of Phannacology and Toxico-S!:7'Olinlversity of Kuopio , p, O. Box 6,

111 21! Kuopio 21 Finland.Ota'. '

O Ilnlcal Museum, University of Oulu, p,l Add ox 191, SF-90101 Oulu, Finland

ress far correspondencc

in Western Europe is an area of increas-ing toxicological significance (2, 3). Sixyears ago the occurrence of a psilocybin-containing species, Psilocybe semilan-ceata (Fr. ex Secr.) Kurnmer , was report-ed (4) in Scandinavia. Chemical and bot-anical work (5) on Norwegian P. semil-anceata showed it to be widcly distribu-ted and a potent hallucinogenic mush-room. Since 1981, the use of psilocybin-containing mushrooms has been record-ed in some narcotic trials in Finland andat least one hospital case has becnreported (6). Therefore we have detcr-rnined , using a high performance liquidchromatographic method, the psilocybinand psilocin contents of P. semilanceatamushrooms picked from various locali-ties in Finland.

Material and Methods

Figure l shows the localities where P. semil-anceata has been found and where sampleswere collected for the present study. Imme-diately after harvesting (August-October1982) the samples were dried overnight at 500

C and then stored in airtight bags at -200 Cuntil analysed. The identities of the driedmushroom sarnples were confirmed by theprofessional rnycologist of our group (E, O,),

References(1) King, R, M" Robinson, H, (1980) Phyto-

logia 46, 446.(2) Bohlmann, F., Zdero, c.. King, R, M.,

Robinson , H, (1983) Liebigs Ann. Chem.2045.

(3) Vichnewski , W., Murari, R., Herz, W.(1979) Phytochemistry 18, 129.

(4) Bohlmann, F" Zdero, c., King, R. M"Robinson, H, (1981) Phytochemistry 20,1657,

(5) Bohlmann, F., Mahanta, p, K., Suwita,A., Suwita , AnI., Natu, A. A" Zdero ,c., Dorner, W" Ehlers, D" Grenz , M.(1978) Phytochemistry 16, 1973.

(6) Seaman, F. C. (1982) The Botanical Re-vie w 48, 121.

The psilocybin and psilocin contents wereanalyzed with high performance liquid chro-matography (HPLC) as described by Chri-stiansen et al. (7) with minor modifications.Each mushroom was analyzed separately.The dried mushrooms were weighed andground to powder which was extracted twicewith a total of 5,00 mI of 10 % l N ammoniumnitrate in methanol. The HPLC system:pump: Varian Model 5000, injector: Rheo-dvne Model 7125 (20 ~I external loop),stationary phase: Hibar® 250-4 (E, Merck,Oarmstadt) prepacked with 5 urn LiChro-sorb", mobile phase: methanol-water-IN01H,NO) (220: 70: IO) buffered to pH 9,6with 2 N amrnonia , flow rate l mI/min. detec-tor: Varian Vari-Chrorn'", wavelength 267mrn, quantitation: pcak areas monitored witha Shimadzu Chromatopac C-RI8 data proces-sar. The calibration graphs from pure psilocy-bin and psilocin standards (Sandoz. Basle )showcd excellent linearity (r 0,9996 and0.9992, rcspectivcly).

Results and Discussion

The recorded distribution of p, semilan-ceata in Finland (Fig. l) suggests that thismushroom can be found in ali parts ofthe country. No clear difference in fre-quency seemcd to occur between thecoastal and inland regions. The analyzcdsamples were found among grass in pas-tures, untrimmed and trimmed lawns, ondecaying straw etc. from 29th August to15th Novernber , though the main seasonin Finland seems to be in September.The phenology of the studied collectionsvaries from the 12th July to the \5th No-vember (in thc years 1867-\(83). Themean dry weight (± S. D.) of a randomsample (100 spccimcns) of Finnish P. se-

278

.20 .60

!77

n

11

KM~", '~'...L.'.~

f"L"41;q-hr-7r'~~~S'-~'------,~-,--i-----'h~~'-j.-·~'----- --'" ,

IO 60AO

Fig.1. Distribution of Psilocybe semilanceata (Fr. ex Secr.) Kumrn.according to the collections made by the authors and the material per-served in the Finnish botanical museums (black dots). The ringed dotsindicate the places of samples analyzed iri the present study.

milanceata was 34.30 ± 18.64 mg. Forthe analyses, one mushroom of mediumsize, one of the smallest and one of thelargest were selected from each location.The average weight loss during the dry-ing procedure was 90 %.

The Iiquid chromatograms showed ty-pically three peaks, one correspondingto psilocybin (rentention volume 6.8

mi), and another (usually very small) psi-locin (LV. 4.7 mi). The third one (r.v. 5.7mi) remained unidentified but is possiblybaeocystein (8).

Psilocin concentrations did not exceed0.02 % of dry weight and were detecr-able only in some samples (detection li-mit about 0.005 %). Psilocybin concen-trations did not seem to correlate with

Pianta Medica 19&;

the date of collection or the geogra h' ,location, and the correlation Wit~ IcaJmushroom size was weaker than ~described from Norwegian P. Serni~ClI]'ceat.a (5). The absol.ute amount of PSij~cybin, however, mcreased with bmushroom size (Table I). l e

'l:'able lo Psilocybin ~ontents and Concentre-

tions In different weight groups of Fin . ,Psilocybe semilanceata (mean ± S.O.) n~_-Dryweight Contenr Concen.

mg mg Iration ~~of dryweigh:

up lo 20 0.27±0.10 1.63 ±OA221-40 0.38 ± 0.07 1.45 ± 0.3341-60 0.73 ± 0.20 1.43 ± O.Y>

over 60 1.20 ± 0.43 1.23 ± 0.30

Mean49.6±36.9 0.64±OAO 1.42±0.36

The psilocybin concentration in theFinnish P. semilanceata varied from0.62 % to 2.37 % (mean 1.42 %) and va-lues of two larger pooled unselectedsamples were 1.68 and 1.53 % of d~weight (i.e. more than l mg/g of freshrnushroom). These values are among thehighest described in any mushrooms.

Acknowledgemenl

Supported in part by the Alcohol ResearcrFoundation, Finland,

References

(1) Pollock, S. H. (1975) J. Psyched. Drur .73.

(2) Francis, r., Murray, W. S. G. (1983) Hu,man ToxicoL 2,349.

(3) Peden, N. R., Pringle, S. D., Crook~.J(1982) Hurnan Toxicol. 1,417.

(4) Schumacher, T. (1976) Va re Nytte1el·ster 71, 110.

(5) Christiansen, A. L., Rasmussen. K.!'H0iland, K. (1981) Pianta Med. ~2.2-~

(6) Haapanen, E., Karjalainen, K., Pelli":':T., Vilska, l. (1983) Suom. Uiiik. \. .'~

2068. E..(7) Christiansen, A. L., Rasmussen. K.'I·'

Tennesen, F. (1981) J. Chromatogr. - l.

163. K E(8) Christiansen, A. L., Rasmussel1. .

(1982) J. Chromatogr. 244, 357.