STRZj2.3

~ybin in Finnish 277

for C)OH4604: 470.340), 452 (19) (M- (C02R); MS: 260.105 (2) (M-RCOzH) (calc. References ~. 424 (9) (452 - COl, 409 (lO) (424 - for ClsH1604: 260.105), 242 (6) (260 - HzO),

!lPJ' _68 (10),246 (18), 205 (58), 187 (51),81 85 (32) (C4H9CO+), 57 (100) (85 - COl; IH_ (1) King, R, M" Robinson, H, (1980) Phyto- C? ';5 (100); addition of diazomethane af- NMR (CDCt), /) ppm): 4.84 (1H, br, d, J = 6, logia 46, 446. ~d the corresponding methylester; IH_ 2-H), 5.73 (1H, br, s, 3-H), 2.61 (1H, br, t, J (2) Bohlmann, F., Zdero, c.. King, R, M., 'Il. (eDCl), Ò ppm): 0.92,0.95,0.98,1.02, = lO, 5-H), 4.69 (1H, dd, J = lO and 9, 6-H), Robinson , H, (1983) Liebigs Ann. Chem. ~ and 3.67 (3H each, s), 4.97 and 4.92 3.18 (1H, dddd, J = 9,4,3,3, 7-H), 5.51 (IH, 2045. ~ IH. br, s), 4.12 (2H, br, s), 2..:..88(lH, ddd, J = 8,8,4, 8-H), 2.91 (1H, ddd, J = 15, (3) Vichnewski , W., Murari, R., Herz, W. ~,J =011. 11,5),2..:..49 (1H, d~d,] - 17, lO, 9,1.5, 9a-H), 1.97 (1H, dd, J = 15, 9, 9~-H), (1979) Phytochemistry 18, 129. 6.35 (1H, d, J = 3.5, 13-H), 5.54 (lH, d,] = 3, (4) Bohlmann, F" Zdero, c., King, R. M" ~.-.'40 (IH. ddd , J - 17,7,4), 13'-H), 2.77 (1H, d, J = 5, 14-H), 2.71 (IH, Robinson, H, (1981) Phytochemistry 20, 589 578 546 436 nm dd, J = 5, 1.5, l4'-H), 2.00 (3H, br, s, 15-H), 1657, . ~ '" _ (CHCt); c = 0.64) 2.30 (1H, tq , J = 7,7, 2'-H), 0.83 (3H, t, J = (5) Bohlmann, F., Mahanta, p, K., Suwita,

The and contents were Psilocybin in Finnish analyzed with high performance liquid chro- matography (HPLC) as described by Chri- stiansen et al. (7) with minor modifications. Psilocybe semilanceata Each was analyzed separately. The dried were weighed and J. Jokiranta I, S. Mustola I, E. Ohenoja ' and M. M. Airaksinen 1.3 ground to powder which was extracted twice with a total of 5,00 mI of 10 % l N ammonium nitrate in methanol. The HPLC system: pump: Varian Model 5000, injector: Rheo- dvne Model 7125 (20 ~I external loop), \bstract: The use of a hallucinogenic mush- in Western Europe is an area of increas- stationary phase: Hibar® 250-4 (E, Merck, ~:\)m.Psilocybe semilanceata, has been occa- ing toxicological significance (2, 3). Six Oarmstadt) prepacked with 5 urn LiChro- sonally reported in Finland, where the spe- years ago the occurrence of a psilocybin- sorb", mobile phase: methanol-water-IN :!S iswidely distributed. We have determin- containing species, Psilocybe semilan- 01H,NO) (220: 70: IO) buffered to pH 9,6 :j, by HPLC. the content of psilocybin and ceata (Fr. ex Secr.) Kurnmer , was report- with 2 N amrnonia , flow rate l mI/min. detec- :'\ilocinin P. semilanceata samples collected tor: Varian Vari-Chrorn'", wavelength 267 ~ùmdifferent parts of Finland; the psilocybin ed (4) in Scandinavia. Chemical and bot- anical work (5) on Norwegian P. semil- mrn, quantitation: pcak areas monitored with ;~ntent was found to be high (0.62-2.37 %, a Shimadzu Chromatopac C-RI8 data proces- anceata showed it to be widcly distribu- ~,ean1.42 % of dry weight). some samples al- sar. The calibration graphs from pure psilocy- " COntained low concentrations (0.01- ted and a potent hallucinogenic mush- bin and psilocin standards (Sandoz. Basle ) % of .02 dry weight) psilocin. room. Since 1981, the use of psilocybin- showcd excellent linearity (r 0,9996 and containing mushrooms has been record- 0.9992, rcspectivcly). ed in some narcotic trials in Finland and Introduction at least one hospital case has becn reported (6). Therefore we have detcr- Results and Discussion The traditional use of psilocybin- rnined , using a high performance liquid :ontaining mushrooms in Centrai Ame- chromatographic method, the psilocybin The recorded distribution of p, semilan- rt\:awas thc focus of intense scientific in- and psilocin contents of P. semilanceata ceata in Finland (Fig. l) suggests that this t ;eres in the 1950s. The psychoactive ef- mushrooms picked from various locali- mushroom can be found in ali parts of :,~ctscf psilocvbin and its active metabo- ties in Finland. the country. No clear difference in fre- ~te, psilocin. ;re similar to those of LSD. quency seemcd to occur between the lisnow known that psilocybin is present coastal and inland regions. The analyzcd notonly in some tropical species, but al- Material and Methods samples were found among grass in pas- :tn some mushroom species in different tures, untrimmed and trimmed lawns, on ns of the world (1). and their ingestion Figure l shows the localities where P. semil- decaying straw etc. from 29th August to anceata has been found and where samples 15th Novernber , though the main season were collected for the present study. Imme- ~------in Finland seems to be in September. loe~art~ent of Phannacology and Toxico- diately after harvesting (August-October 0 The phenology of the studied collections S!:7'Olinlversity of Kuopio , p, O. Box 6, 1982) the samples were dried overnight at 50 111 21! Kuopio 21 Finland. C and then stored in airtight bags at -200 C varies from the 12th July to the \5th No- Ota'. ' O Ilnlcal Museum, University of Oulu, p, until analysed. The identities of the dried vember (in thc years 1867-\(83). The l Add ox 191, SF-90101 Oulu, Finland mushroom sarnples were confirmed by the mean dry weight (± S. D.) of a random ress far correspondencc professional rnycologist of our group (E, O,), sample (100 spccimcns) of Finnish P. se- 278 Pianta Medica 19&;

.20 .60 the date of collection or the geogra h' , ! location, and the correlation Wit~IcaJ mushroom size was weaker than ~ 77 described from Norwegian P. Serni~ClI]' ceat.a (5). The absol.ute amount of PSij~ cybin, however, mcreased with b mushroom 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 ±OA2 21-40 0.38 ± 0.07 1.45 ± 0.33 41-60 0.73 ± 0.20 1.43 ± O.Y> over 60 1.20 ± 0.43 1.23 ± 0.30

n Mean49.6±36.9 0.64±OAO 1.42±0.36

The psilocybin concentration in the 11 Finnish P. semilanceata varied from 0.62 % to 2.37 % (mean 1.42 %) and va- lues of two larger pooled unselected samples were 1.68 and 1.53 % of d~ weight (i.e. more than l mg/g of fresh rnushroom). These values are among the highest described in any mushrooms.

Acknowledgemenl

Supported in part by the Alcohol Researcr Foundation, Finland,

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

(1) Pollock, S. H. (1975) J. Psyched. Drur . IO AO 60 73. Fig.1. Distribution of Psilocybe semilanceata (Fr. ex Secr.) Kumrn. (2) Francis, r., Murray, W. S. G. (1983) Hu, according to the collections made by the authors and the material per- man ToxicoL 2,349. served in the Finnish botanical museums (black dots). The ringed dots (3) Peden, N. R., Pringle, S. D., Crook~.J indicate the places of samples analyzed iri the present study. (1982) Hurnan Toxicol. 1,417. (4) Schumacher, T. (1976) Va re Nytte1el· ster 71, 110. milanceata was 34.30 ± 18.64 mg. For mi), and another (usually very small) psi- (5) Christiansen, A. L., Rasmussen. K.!' the analyses, one mushroom of medium locin (LV. 4.7 mi). The third one (r.v. 5.7 H0iland, K. (1981) Pianta Med. ~2.2-~ size, one of the smallest and one of the mi) remained unidentified but is possibly (6) Haapanen, E., Karjalainen, K., Pelli":': T., Vilska, l. (1983) Suom. Uiiik. \. .'~ largest were selected from each location. baeocystein (8). The average weight loss during the dry- Psilocin concentrations did not exceed 2068. E.. (7) Christiansen, A. L., Rasmussen. K.'I·' ing procedure was 90 %. 0.02 % of dry weight and were detecr- Tennesen, F. (1981) J. Chromatogr. - l. The Iiquid chromatograms showed ty- able only in some samples (detection li- 163. K E pically three peaks, one corresponding mit about 0.005 %). Psilocybin concen- (8) Christiansen, A. L., Rasmussel1. . to psilocybin (rentention volume 6.8 trations did not seem to correlate with (1982) J. Chromatogr. 244, 357.