J~P HA Rt1'. s c I. I ~ I it"3 - l f.41 (l C) 11) Baeocystin in Psilocybe semilanceata DAVID B. REPKE *x and DALE THOMAS LESLIE t ---- --------------- ---,_._---- .\h'l r act o Ba-ocvstin and psilucvhin were found in extracts of yloxy<~-(2-methylaminoethyl)indole and its demethyl .' .•ilrJ!'lv 01'Psilocvbc scmilanceuiu Psilocybin (but not baeocystinl counterpart (baeocystin and norbaeocystin, respec- wa~ also detected in extracts of a related species, Psilocybe pellicuìosa. tively) have been reported only in P. baeocystis Singer Traces of psilocin were present in these two species. The structures -and Smith (13). These compounds were not found in P. ••r Ihp isolated cornpounds were corroborated using mass spectrornetry attrobrunnea (Lasch) Gillet, P. pelliculosa (Smith) al1(: ll\' spectroscopv. Singer and Smith, P caerulipes (Peck) Sa cc., or P. h",\phrases O Haeocvstin-c-isolated from extracts of Psilocvbe «trictipes Singer and Smith (14). " "lIlon(,f'Ola, identified hv rnass spectrometry and UV spectroscopy During investigations of the indole alkaloids of the O Psilocvbin=isolated from extracts of Psilocvbe semilonceota and J' pcllicu {oso, identified by rnass spectrometry and UV spectroscopy genus Psilocvbe. a compound with a mobility slightly O I'silocvbc semi/oneeota--extracts of carpophores. baeoeystin and slower than psilocybin was consistently detected on J"iloc"hin isolated. identified bv mass spectrometry and UV spec- thin-layer chromatograms of extracts of a Pacific t nt~('()JlY Northwest varietv of P scmilonceat a This compound was observed in ~ll collections of this species examined The genus Psilocybe contains many species with over several years. hallucinogenic properties (1). Their use for religious purposes in Mexico was first recorded 4 centuries ago EXPERIMENTAL' (:21.However, only in the last 35 years has serious study !wen devoted to this subject (3l. In 1959, two active Fn-ezr--dned carpophore-, of V srrnitaucrut a C',H24 nlg. were grpund principles were isolated from several members of this t.u a powder and extracu-d hv shakirn; with rnet hanol 81 roorn tern genus (4): the 4-hydroxy-3-(2-dimethylaminoethyl)- indole (psilocin) and its corresponding phosphate ester " 1 TLC wa-, carried out u-",inJ! 0.2;\- and 1.(1. rum 18.\'{"r~ or fOi1iCHJ!pl (; lo' on da~" I psilocybin). Since then psilocin and psilocybin have pla1t"!'. Thp solvr-nt svstr-m tI~-Ò wa- I-pr,.pantll ~.,. arnmonium hH1ruXlft.· l:l:~1 Il:~1. Ma sJw{'lril 'At'fI' 11lll~JilH'd wit h an Ali" .. CH--1 ~pt'CirltnU'll·~ .• " d!;~,\-' iWE'n detected in other mernbers of Psilocvbc and re- inìr-i. Ttu- pn.h! 1t-nqwraT~:I' "'l'', :J,d~'~.HlId H," l'''' ~1I\Jrn' pnlPnTl:t! "d" ';"1' t-v !diPd genera (!)), . A ( .cH\ modt, j l ~,~Hh'l n ,I) n, ", .me: t'T" V. ,j~lI,,~'(J i, o ckll'fmint· l'\ ...: )t·(·t ~.: - 'l'11f' ("\l!iI·ctl(\!)~ ar- IL tht· h-r hanun (,j lht E~(,lIda !\IH'lon:l: d« ~'It'III'ld~ Heirn (6) sugzested that the hallucinogenic mush- Bj,d,·,:':I("H~. I P ,. "tlUfP Il. t' .. "1t·):Il"t •. iillei -,, ,nlt' Ufi a 1:-.,I rm (h-I'(.~il ;!1 tt« r"',m described in a lfith centurv English re pori was h(-rllarlllrn 01 tht ':IlI\t·r"ll\ 101 Mh'ni;:ar: Ann Arbor . ~11('h C .••rVllpÌA . .-,-.. (,j l' ~."iilfJljc('nto \lt,"f'rr ,-,,:Jt--cl{·n In pa:-ltJrt'~ al t t l(' lollowlI1r h)(·.·tl-,1)IJ~ v-ru v- J'sl!ocybe semilanceaia (Fr.I Kumrner (Stropharia- Harbor L'ount v. \'a,h, J!C:.' J!!~:, ILE:-;UE J:l;>1 ì alsoat MWHI, JNi~ 'ai", ~'eae considered t.he tvpe species of the genus (7, 8\. MI !\,lH'HI,:.'~:.'I;t81,.'Al M!(,H, and:.'h,CI:,Jeff~NmC",m" \'a.,h .. l!!::: I!C, J. 11.1':.-:1.1F l N:: ",I.,,, A: \11<'H " :.'~'''' ,al", Al M !C'H l, a nd :."~·IL Ha nd;,·. \, iI,l. , I rJ\ p,t igator~ ha\'(' verifir-d Ihf' prp"éf)(T or psilorvhin 1"7' !1.!-->~l.1F~·-:-\II!';'!'(~:--:!Xt..·..()rt l~j~·:.II.J-:"I.IF_",~! \·,.;.,II"':t~l't,l i" . .",. """:. w e-rr j.~:;:IC l!. i,.,.···..!...,.n "",,toIod cit,!,q .. ;11H~"'.'''''::!- ,l'i ,L', ~ h.:!I.,'· 1';,1:,11(1\ psilocin : in ,.;n't'ra! ElIropean ('01It".'t ion- 01 l' (,.,\,,;\ \\;j"';'l: l~'-' ;:.L~:.lt" ì-· .' And \LI\~':\"L \I.:I"! 1""" :l.t-: .....:.lr- , ,iunccala (~L 1:21. :..-.:•,..•.'!'rl~ 1.1\lll. .:il"-;~-:d;,·i~ li,.r· d"! .,..,..: lo rr: . l . P:II1"·;'''·c df'~'; T"IP, Il '11"" p; l ltt Ll:"'lw~in!' •., ",;:';'/( ,'(./(; 11\;: ....~'nlt \l,,·d.·, t e tu.nn- 1"1'1' T1Jf presence of the psilocyhin analogs 4-phusphur- n':lllll~' 1111"~pt-\'dH pi:tct'Il.' Il: winc l. t urtlnr ...\l. Hl\ fllii~ \ ia nt . l" fid,.', '''r :!ii hr l'h, rr..xt ur« WH' filtered, and th« !.nhd~ were RITEHENCES ,,,,,l'''d Wl' h me-t !.ltnol Th- rorn hme-d filtrate and washings were "onu'lil ra t ••d '" l'!lI'Uf, Analvt ical TLC of the extract (tw« develop ( l r R Herrr. and R G. Wassun, "Les i 'hilmpignon; Ha llurino rru-ntx) ,h"w~d psilo'ybm (RI 0.27) and the slower cornpound (RI g-f.nesdu Mr-xique." Museurn National D'Histoire Naturelh , l'ano, O.LL) Both spol!. r-xhibited the same dark-blue coloration when France.1959. slHliV~d with 2'lo. 4·dlmethylaminobem..aldehyde in ethanol and ex- (21 B. de Sahagùn, "Fiorentine Codex, Generai History of New pose-d ti, hydrogen chloride vapors. Trace amounts of psilocin were Spain," Books III and XI, A. J. O. Anderson and C. E. Dibble, also detected. Translators, University of Utah Presso Salt Lake City, Utah, 1970. The concentrated residue was preparatively chromatographed on (3) R. G. Wasson, Bot. Mus. Leatt. Haro. Univ., 20,25(1963). a lumm layer of silica gel in the described solvent system. A band (4) A. Hofmann, R. Heim, A. Brsck. H. Kobel, A. Frey, H. ou. rontaining the two componenti! W8& removed from the plate, and the T. Petrzilka, and F. Troxler, Helv Chim. Acta, 42,1557(1959). cornpounds were eluted with methanol containing 5% aqueous am- (5) R. E. Schultes and A. Hofrnann, "The Botany and Chemistry rnonia. The eluate was concentrated under reduced pressure, and the of Hallucinogens," Charles C Thomas, Springfield, Ill., 1973, pp. residue was rechromatographed on a 0.25-mm layer of silica gel to 36-52. resolve t he two cornponents. Two developmentB in the propanol- - (6) R. Heim, Natur. Can., 98,415 (971). amrnonia system followed by elution of the bands afforded psilocybin (7) R. Singer, "The Agaricales in Modern Taxonomy," 2nd ed., Cl rng. (Ufi<Jul, mp 175-180° [lit. !l5) mp 185-195°1 and its slower- J. Cramer, Weinheim, Germany, 1962, p. 542. movirig counterpart (J mg. 0.12%), mp 245-248° [Iit. (14) mp 254- (8) P. Konrad and A. Mauhlanc, "Rèvision des Hyménomycètes 2;)1'°1 Carpophores of P pelliculosa were extrarted in a similar de France," Paul LeChevalier. Paris, Franee, 19~7, p. 84. manm-r lo giv« ()'()8'\,psilocvhin Traces of psilocin were also detect- - (9) A. Hofmann, R. Heim, and H. Tscherter, C. R. Acad. Sci., 257, f'd 10(1963). :10) R. G. Benedict, V. E. Tyler, and R. Watling, Lloydia, 30, RESUL TS AND DISCUSSION 150(967). (11) P. G. Mantle and E. S. Waight, Trans. Br. Mycol. Soc .. 53, The UV spectra of the two compounds were superimposable, with 302(1969). À",", (methanol) 290 (. 4000), 280 (sh)(5000), 268 (6300), and 222 (12) M. Semerdzieva and F. Nerud, Ceska Myko/., 27,42(]973). (40,000) nm, typical ofpsilocybin (4). The mass spectra were consis- (13) A. Y. Leung and A. G. Paul, J. Pharm. Sci., 57,1667(1968) tent with published data for psilocybin (11) and baeocystin (13). (l4) lbid, 56,146(1967). Psilocybin showed peaks at m/e 204 (relative abundance, 19%), 160 (15) A. Hofmann, R. Heim, A. Brack, and H. Kobel, EIperientia, (4).159 (3), 146 (6),130 (3),117 (2), and 58 (100). Baeocystin showed U, 107Cl958). peaks at 190 (8),160 (4),159 (4),147 (16), 146 (14), 130 (4),117 (5), (6) H. Budzikiewicz, C. Djerassi, and D. H. Williams, "Structural and 44 (JOO). Elucidation of Natural Products by Mass Spectrometry," vol. l, Each compound exhibited typical fragrnentations of 3-indo- Holden-Day, San Francisco, Calif., 1964, pp. 42-45. lylethylamines (16), e.g., a base peak arising from tl-bond fission of (17) V. E. Tyler, Lloydia, 24, 7W96l). the ethylamine side chain (psilocybin, m/e 58; and baeocystin, m/e (18) R. G. Benedict, in "Microbial Toxins," S. Kadis, A. Ciegler, 44 l. While neither compound showed a parent peak, both showed and S. J. Ajl. Eds., Academic, New York, N.Y., 1972, chap. Il. sianals for the dephosphorylated species (psilocybin, m/e 204; and (191 S. I. Stein, G. L. Closs, and N. W. Gabel, Mvcupathol. Myco/ baeocystin, mie 190). The concentration of psilocybin in P. semi- App., Il, 20;,(1959). lanccato was consistent with isolated yields reported from European collect ions (9,11). ACKNOWLEDGMENTS AND ADDRESSES In agreernent with Leung and Paul (14), baeocystin W8& not de- tected in P. pel/icu/osa. Psilocybin (but not psilocin) has been re- Received December 29, 1975, from • 148-B West Dona Street , ported previously in this species (17). Mountain View, CA 94041. and IlU4 Whitney A"enue, Los Gatos, Preliminary investigations suggest the presence of baeocystin in CA 95030 ot her species and genera.