June, 1943
ISOLATION AND STRCCTURES OF SOMENEWSTEROIDAL SAPOGENINS
[CONTRIBUTION FROM
THE
1199
SCHOOL OF CHEMISTRY AND PHYSICS OF THEPENNSYLVANIA STATE COLLEGE]
Sterols. CLVII. Sapogenins. LXM.' Isolation and Structures of Thirteen New Steroidal Sapogenins. New Sources for Known Sapogenins BY RUSSELLE. MARKER,R. B. WAGNER,PAUL R. ULSHAFER,EMERSONL. WITTBECKER, DALEP. J . GOLDSMITH A N D CLARENCE H. RUOF
Due to the pressure of war work, the publica- Frank C. Whitmore and Dr. Oliver Kamm for tion of our studies on the sapogenins of over three their encouragement in this project. We thank hundred plants from Mexico and United States Dr. H. M. Crooks and Dr. G. H. Fleming. We will necessarily be delayed. In the present paper thank Parke, Davis and Company for their aswe are reporting some of our results in preliminary sistance. form . New Steroidal Sapogenins Earlier work on the biogenesis of the steroidal 1. Hecogenin.-In the course of our plant hormones and related compounds has focused our studies, we first isolated from Hechtia texensis (S. attention on this problem in both animals2 and Wats.) a new steroidal sapogenin having the complants. In the present paper we report the isola- position C27H4204, m. p. 245', 253' and 268', tion and structures of thirteen new steroidal sapo- which we have named hecogenin (I). Anal. genins, thus more than doubling the number of Calcd. for C27H4204: C, 75.3; H, 9.8. Found: these substances known. We have also found C, i 5 . 5 ; H, 9.9. When refluxed with acetic annew sources for many of the previously known hydride, it forms a monoacetate having two forms, sapogenins. m. p. 243 and 252'. Anal. Calcd. for C29H4406: The extensive collections by the senior author C, 73.7; H, 9.4. Found: C, 73.9; H, 9.3. Alwere made possible only because of the kind and though hecogenin (I) forms a monosemicarbapainstaking cooperation of the Botany Depart- zone, it is unaffected by the conditions of a mild ments, Experimental Stations and Range Ecology Clemmensen reaction, indicating the inert posiDepartments of the following Institutions : Texas tion of the carbonyl group. Wolff-Kishner reducA. and M. College, University of Arizona, Uni- tion removes the carbonyl group giving tigogenin versity of California, New Mexico State College, (111, m. p. and mixed m.p., 206'. Anal. Calcd. North Carolina State College, Coker College, for C27H4403: C, 77.8; H, 10.6. Found: C, University of Mexico, Sol Russ College, Uni- 77.5; H, 10.4. Vigorous oxidation with chromic versity of Florida, Wac0 College, Purdy Botanical anhydride in acetic acid gives hecogenic acid Gardens (Ukiah, Calif.), Laredo Cactus Gardens (111),m. p. 268' dec. Anal. Calcd. for C27H40(Laredo, Texas), Missouri Botanical Gardens, 0 1 : C, 68.0; 8.4. Found: C, 68.3; H I 8.3. The and Huntington Botanical Gardens (San Marho, latter is not identical with chlorogenic acid (IX), Calif.). We thank Dr. Lester Mallory and Mr. digitogenic acid or digitoic acid. Furthermore, Douglas Crawford, United States Embassy, the dimethyl ester of 111, m. p. 18f0,does not corMexico City, Ing. Eduardo Morillo Safa, oficial respond to any of the dimethyl esters of the above Mayor Departamento de Agricultura y Fomento, acids. Mild oxidation of hecogenin (I) with Mexico City, and The Ministry of Public Health of chromic anhydride gives hecogenone (IV), m. p. Mexico for making the Mexican collections pos- 240'. Anal. Calcd. for C27H4004: C, 75.6; H , sible. We thank especially the following botanists 9.5. Found: C, 75.3; H, 9.2. The latter is not (listed alphabetically) for aiding in the collection identical with either chlorogenone or T-ketotigoand identification of the plants: Dr. G. Bene- genone prepared from diosgenin. We propose vides, Dr. Lyman Benson, Dr. A. B. Conner, Dr. structure I for hecogenin. and Mrs. V. L. Cory, Dr. Ladd Cuitak, Dr. C. J. It is noteworthy that no previously known steroEpling, Dr. A. B. Hershey, Dr. Wm. Hertrick, idal sapogenin having the spiro-ketal side-chain Dr. H. Lewis, Dr. H. B. Parks, Dr. L. N. Pultz, and established structure has a carbonyl group. Dr. B. E. Smith, Dr. 0. T. Sperry, and Dr. J. J. We have processed on an average of one hunThornber. We express our appreciation to Dean dred pounds of each species in the accompanying (1) Far previous papers in this series see Sterols. CLVI Sapolists in the manner described b e f ~ r e . ~The regenins. LXVIII, THISJOURNAL, 65, 1248 (1943). (2) Marker, ibid., 60,1725 (1938).
(3) Market, Wagner and Tlehafer. r b z d , 64, 1283 (1942)
1200
MARKER, WAGNER, ULSHAFER, WITTBECKER, GOLDSMITH A N D RUOF
c
Vo1. 65
c
TI. Tigogeriin
c I
111. Hecogenic Acid
sults and geographical locations of the plants =e given. The other sources of hecogenin, irnmediately following, make this substance readily available.
IT'.
.c-c,
Hecogenone
tures and the analyses never varied more than three-tenths from the calculated carbon and hydrogen values. 2. Manogenin.-We have isolated from X a n Hecogenin, freda maculosa (Hook.) a new keto-steroidal sapoR. per kg Plant T.,ocation (dry) genin, having the composition C27H4205, m. p. San Antonio, ' r e x . Agasi americana (L.) 0.4 '711-243', which we have named rnanogenin. A . americano war. Tucson, Ariz. 3 .0 A . asperrima (Jacobi) Monterey, Mexico 0.1 Anal. Calcd. for G~H4205: C, 72.6; H, 9.5. Devils Canyon, Ark. A . cvysantha (Peebles) 1.2 Found: C, T2.3; H , 9.4. Boiling acetic anhyA . deserti (Engelm.) Palm Springs, Calif. 0 G A . deserti var. Goldroad, Ariz. 3 .3 dride formed a diacetate, rn. p. 255'. Anal. Orizaba, Mexico A . endlichiana (Trel.) 0 .6 Cdcd. for C41H4607: C, 70.2; H, 5.8. Found: C, Tucson, Ariz. A . expansa 2.0 A . fourcroydes (Baker) Victoria, Mexico 0.1 70.2; H, 8.6. The function of four of the five El Capitan Mt., Tex. A . gracilipes (Trel.) 7 0 oxygens is shown by its conversion by the WolffA . huachucensis (Baker) Fort Huachuca, Ariz. (1.3 A . murpheyi (Gibson) Globe, Ariz. I .0 Kishner method to gitogenin (VI), m. p. and 0 2 Tucson, Ariz. A . palmeri (Engelm.) mixed ni. p., 2BK-2(iS0. A n d . Calcd. for CZ7Hd4Nogales, Ariz. A . Purviflora (Torr.) 3 0 5 Lower California, Mexico A . shawii (Engelm.) 01: C, i t . 9 ; H, 10.2. Found: C , 73.2; H, 10.1. Superior, Ariz. A . foumeyana (Trel.) 13 0 Xanogenin (V)upon mild oxidation with chromic Hechfia texcnsis ( S . Wats.) Big Bend, Tex. 0 1 Hesperaloe funifera (Koch) Huasteca Canyon, Mexico 1.3 mhydride in acetic acid gave hecogenic acid Maguey el ojifal E1 Ojital, Mexico iil . 1 (III), 111. p. and mixed in. p., 268' dec. Anal. !I 5 Zitacuaro, Mexico Maguey espadin Maguey teguilla manso 11 1 Tanaquila, Mexico Calcd. For C?;HrliOi. C , 68.0; H, 8.4. Found. :i / I San Antonio. Te.: Mnnfvedu maculosa (Hook C. 68.3; H, 8.3. The identity was further esThe identity of the hecogenin was established tablished by the direct comparison of the diin each case by analysis of the genin and its ace- inethyl esters, m. p. and mixed m. p., 186-187'. tate along with mixed melting point determina- The carboiiyl group in manogenin like that in tions on both. Hecogenin has three polymorphic hecogenin cannot be removed under the condiforms melting a t 245, 253 and 268', and its ace- tions of a mild Clenirnensen reaction. TVe protate, resembling sarsasapogenin acetate and pose structure V for manogenin. smilagenin acetate, has two forms, ni. p. 243 and Although rnanogenin occurs abuiidantly in 252'. In all cases the melting points and mixed J1anfP.edu maculosu (Hook.), the latter itself is melting points of the genin and acetate were not scarce. We have found twenty-four additional more than three degrees below the above tempera- sources, listed in the following table.
ISOLATION AND STRUCTURES OF SOMENEWSTEROIDAL SAPOGENINS
June, 1943
e
1201
ing the course of our investigation of the Yucca for steroidal sapogenins, we have isolated from Y. schottii (Engelm.), Y . data (Engelm.) and Y . 4 i:accida (Haw.) a new steroidal sapogenin havHO/ ing the composition V. Manogenin VI. Gitogenin C27H4204, m. p. 246' and JCr03 ( 2 5 T ) 232', which we have 111. Hecogenic Acid named yuccagenin (VII). ManoAnal. Calcd. for C27H4204: d, T3.3; H , 9.8. genin, g. per kg Found: C, 75.3; H, 9.8. Boiling acetic anhydride Location Plants (dry) formed a diacetate, m. p. 1%'. Anal. Calcd. for Tlaxcala, Mexico 0.1 Agave atrovirens (Otto) 1.7 Huasteca Canyon, Mexico A . bracteosa (Wats.) C31H4606:C, 72.3; H, 9.0. Found: C, 72.4; H, 0 7 Big Bend, Tex. A . chisoensis 9.0. Catalytic hydrogenation (Xdams catalyst) of 0.9 Bueno Vista, Mexico A . crassispina (Trel.) Portezuelo, Mexico 1.7 A , f e r o x (Koch) yuccagenin (VII) in ether containing several drops El Capitan Mt., l'ex. 0.6 A . gracilipes (Engelm.) of acetic acid gave gitogenin (VI), m. p. and mixed 0 2 Davis Mts., T e x . A . havardiana (Trel.) Fort Huachucu. hriz. 5.3 A . huachucensis (Baker) m. p., 264-266'. Anal. Calcd. for C2iH~04:C, Portezuelo, hIexico 0 8 A . lehmanii 74.9; H, 10.2. Found: C, 7 3 . 2 ; H, 10.0. TreatMonterey Mexico 0.5 A . lophanlha 0.3 Tacuro, Mexico A . mirabalis (Trel.) ment of yuccagenin (VII) with hydrogen peroxide Tehuacdn, Mexico 0.6 A . initraeformis (Trel.) in acetic acid gave a tetrol (VIII), m. p. 350'. Parras, Mexico 0.8 A . 9arassana (Trel.) 0.3 Saltillo, Mexico A . puiotefera (Trel.) Anal. Calcd. for C~H4406: C, 69.8; H, 9.5. 1.1 Portequelo, Mexico A . salmiana (Otto) Found: C, 69.4; H, 9.6. The latter upon mild Iron Mt., Tex. 0.2 A . scabra 1 .o Ixmilquilpan, Mexico A . striata (Zucc.) oxidation with chromic anhydride in acetic acid 1.3 St. George, Utah A . ufahensis (Engelm.) followed by dehydration and zinc-acetic acid re0.5 Orizaba, Mexico Maguey cacaya 0.8 Jacala, Mexico M. canaslo duction gave chlorogenic acid (IX), m. p. and 1.2 Zitacuaro, Mexico M. ceniso mixed m. p. 233-234'. Anal. Calcd. for (2271.0 Zitacuaro, Mexico M. cimmarron M . cuchacamba Tanaquilla, Mexico 0.5 Hd007: C, -68.0; H, 8.4. Found: C, 68.1; H , ik'anfreda tigrina (Engelm.) Bonneau, S. Car. 0.3 8.5. The identity was further established by the The identity of the manogenin was established direct comparison of the dimethyl esters, m. p. in each case by analysis C C I /c-c, I c-c of the genin and its acec-c( - ic-c c-c( >c-c tate along with mixed C I I O C melting point determinaA I A / O tions on both. Manoo-c genin, resembling hecogenin, has three polymorphic forms melting a t 243, 254 and 264', and its aceOH tate, resembling sarsa- y11, Yuccageniii VI. Gitogenin XXXI. Chlorogeniri C sapogenin acetate and I C c-c smilagenin acetate, has three forms melting a t 215, 242 and 255'. In all cases the analyses never varied more than three-tenths from the --t dehydration and reduction calculated carbon and ' O H hydrogen values. OH 3. Yuccagenin-Dur~ 1 1 1 . Tcatrol IX. Chlorogeiiic Acid
c-c
C
I
c-c /c-c \c-c \'-c/
pyy-: ChA
4
MARKER, WAGXER, ULSHAFER, WITTBECKER, GOLDSMITH AND RUOF
I202
aiid mixed m. p., 163-1(2". U'e propose structure l r I I for yuccageniii. 4. Kammogenin.-We have described above the isolation of yuccagenin (VII) from Yucca sc-hottii (Engelm.) which accounted for the major portion of the total steroidal sapogenin fraction. From the mother liquors of the crystalline yuccageiiin diacetate we obtaiiied d new steroidal sapogenin having the cornposition C27H.1006, 111. p. 242". Annl. Calcd. for C2iH4,,OS. C, 78.9; H , !).I. Found: C. i'2.(\. H, 9.1. We have found other sources of this new substance, namely, Samuela carnerosann (Trel.), Yucca brevifolia (Engelm.) and Yucca harrimanii (Trel.). Boiling acetic anhydride formed a diacetate having two forms, 111. 1'. 2.13' atid "li0'. tlnul. Calcd. for C,jlH4407: C, 70.4; H ,
MARKER, WAGNER,UISHAFER,WITTBECKER, GOLDSMITH AND RUOF
1204
c
C
Vol. 6.5
10.2. Found: C, 74.9; H, 10.3. The identity "-c< >c-c of this degradation C A !, o--c product (XV) was further established by its conversion to samoPiaOEt genin diacetate, m. p. and mixed m. p., 195HO' 198'. Anal. Calcd. Xi'I. Epd-samogenixi XV. Samogenin for C31H4806: C, 72.1; ' CrOI (25"i H, 9.4. Found: C, C I c-c 72.0; H, 9.4. We sugc-c< >c-c gest structure XIX for c , I 0-c mexogenin. 10. YamogenhWe have obtained from Dioscoreu testdinaria I I HOOC"/d HO0Cv/J in good yields diosgenin, H H isolated as the acetate, XVIII. Gitogenic acid XVII. Samogenic acid m. p. and mixed m. p., Anal. Calcd. for-CZ9H4404: C, 7613; accounted for the major portion of the total ster- 199-202'. oidal sapogenin fraction. From the mother liquor H, 9.7. Found: C, 76.1; H , 9.5, Hydrolysis of the crystalline samogenin was obtained a new of the acetate gave diosgenin, m. p. and mixed Anal. Calcd. for C Z ~ H ~ Z O ~ : steroidal sapogeiiin, isomeric with manogenin (V), m. p., 206-209'. melting 246'. Anal. Calcd. for C Z ~ H ~ OC,~ : C, 78.2; H, 10.2. Found: C, '77.8; H, 9.9. 72.6; H, 9.5. Found: C,72.9; H, 9.6. It also From the mother liquors of the crystalline diosoccurs in the steroidal fraction from Yucca schottii genin acetate was obtained a substance, isomeric Anal. (Engelm.). Boiling acetic anhydride formed a di- with diosgenin acetate, melting 180-182'. acetate, isomeric with manogenin diacetate, melt- Calcd. for CzlH@04: C, 76.3; Jff, 9.7. Found: ing 208'. Anal. Calcd. for C31H460,: C, 70.2; C, 76.3; H, 9.8. A mixture with diosgenin aceHydrolysis of this maH, 8.8. Found: C, 70.3; H, 9.0. We propose tate melted 164-167'. terial gave the free genin, isomeric with diosgenin, the name mexogenin for the new substance. Although mexogenin (XIX) forms a monosemi- melting 2w201'. Anal. Calcd. for C27H4203: carbazone, i t is unaffected by the conditions of a C, 78.2; H, 10.2. Found: C, 78.1; H, 10.3. mild Clemmensen reaction, indicating the inert We have named this new steroidal sapogenin, position of the carbonyl group. In this respect, yamogenin. Treatment of yamogenin (XX) under the usual it is like the other 12-keto-steroidal sapogenins, namely, hecogenin (I), manogenin (V), furcogenin conditions employed for the isomerization of the (XIII) and kammogenin (X). WoH-Kishner sapogenin side-chain4 gave diosgenin (XXI), reduction removes the carbonyl group, giving m. p. and mixed m. p., 206'. Catalytic hydrosamogenin (XV), m. p. and mixed m. p., 210- genation (Adams catalyst) of yamogenin acetate 212'. AnaE. Calcd. for C27H4404: C, 74.9; H, in ether containing several drops of acetic acid gave neotigogenin acetate (XXII), m. p. and mixed m. p., 179-180'. Anal. Calcd. for C)z9H4604: C, 75.9; H, 10.4. Found: C, 76.0; H, 10.3. A mixture with the starting material melted 171173'. The identity of XXII was further established by a direct comparison of the hydrolyzed Ho\~~/"$/----' Wolff-Kishner Samogenin product, m. p. and mixed m. p. with neotigogenin, 202-203'. XV Anal. Calcd. for C27H4403: C,77.8; HO/ H, 10.6. Found: C, 77.8; H, 10.6. We propose structure XX for yamogenin. XIX. Mexogenin C-C
I
1
_____f
ClJ
ISOLATION AND STRUCTURES OF SOME NEWSTEROIDAL SAPOGENINS
June, 1943
1205
acid a t the boiling point gave samogenin (XV), m. p. and mixed m. p., 2100 0 212'. Anal. Calcd. for C27Hu04: C, 74.9; H , HCl-EtOH 10.2. Found: C, 74.8; H I 10.1. Thus, it differs 1 day from samogenin (XV) only in the configuration of XX. Yamogenin XXI. Diosgenin the side-chain. In a like manner, sarsasapogenin (XXX) differs from smilagenin (XIV) ; the former can also be converted to the latter by ethanolic hydrochloric acid.j We proC l I I pose structure XXIII for texogenin. 12. Nologenin.-Recently,6a we reported the XXII. Neotigogenin isolation of nologenin, a We have found twelve new sources for yamo- new steroidal sapogenin, from Beth root, having the composition C~H4406,m. p. 265'. Four of genin, listed below under diosgenin. 11. Texogenh-We have processed 1364 kg. c HO of Yucca schottii (Engelm.). We have found that yuccagenin (VII) accounted for 59% of the total crystalline sapogenin fraction. From its mother liquors was obtained smilagenin (XIV) (13% of the sapogenins), kammogenin (X) (13y0 of the HCl-EtOH sapogenins), samogenin (XV) (S% of the sapo1 hr. genins), gitogenin (VI) (2% of the sapogenins), HO/ mexogenin (XIX) (1y0 of the sapogenins) and a XXIV. Nologenin C new steroidal sapogenin, isomeric with samogenin I c-c (XV), melting 171-172', which we name texoHoc-c( )c-c genin. Anal. Calcd. for C27H4404: C, 74.9; o-c H, 10.2. Found: C, 74.7; H, 10.1. The yield I of the latter was 4s/o of the total sapogenin fraction. Boiling acetic anhydride formed a diaceHCl-EtOH + tate, isomeric with samogenin diacetate, melting 2 hr 170-172'. Anal. Calcd. for CilHtBOa: C, 72.1; H d XXV. Pennogenin HI 9.4. Found: C, 71.8; H I 9.3. Treatment of c o texogenin (XXIII) with ethanolic hydrochloric C
C
C-C
r
,&Lp XXIII.
Texogeniii
H O / e V ' XXVI.
Kryptogenin
(51 Marker and Rohrmann, THISJOURNAL, 61, 896 1939). (6) (a) Marker and co-workers, i b i d . , 66, 1248 (1943); (b) 66, 739 (1943).
MARKER, WAGNER, ULSHAFER, WITTBECKER, GOLDSMITH AND RUOF
1206
c
HO/
Vol. 65
c
'u k"
XXVII. Kappogenin
I
l
Hydrolyii,
I
c
I
I
"2-EtOII 9U niiti.
I
HO/\/V
:I. Diosgenin
LXIX
the five oxygens are present as hydroxyl groups, two of which are acetylated with boiling acetic anhydride forming a diacetate (C31H4807),rn. p. 200'. Mild treatment of nologenin (XXIV) with ethanolic hydrochloric acid gives pennogenin (XXV)@"isolated as the monoacetate, m. p. and mixed rn. p., 198-199'. Anal. Calcd. for (229Hd&&' C, 73.7; H, 9.4. Found: C, 73,s; H, 9.4. Prolonged treatment of this product (XXV) with dianolic hydrochloric acid gives kryptogenin (XXI-I).Rh wc propose structure XXIV for iiologenin. 13. Kappogenin.--We have reported the isolation and structures of two new steroidal sapogenins from Beth root (Trillium erectum), namely, nologenin"' and kryptogeninSfi"These are peculiar in having an open side-chain instead of the usual spiro-ketal side-chain possessed by the other sapogenins. Furthermore, their close structural relationship is shown by the conversion of the former to the latter by prolonged ethanolic hydrochloric acid treatment. We h a w isolated from the mother liquor of nologenin (XX IV ) another sapogenin having an open side-chain, which we have named kappogenin, rn. p. 230'. Anal. Calcd. for C27H4401: C, i4.9; H, 10.3. Found: C, 75.0; H, 10.4. T w o of the three hydroxyl groups react with boiling acetic anhydride forming a diacetate, ni. p. A n d . Calcd. for C31H4806:C, 72.0; H, !I L Izorind: C, 72.2; H, !?.~4. Treatment o f !,appogrtiiri iSS\'TI
I
foi
iiiiic.t\
niinri(t~\
ivitli
.i,ltj-pregiiadien-3(B)-ol-20-one
boiling 2 N ethanolic hydrochloric acid gives diosgenin (XXI) isolated as the monoacetate, rn. p. and mixed ni. p., 198-200'. Anal. Calcd. for C29H4404: C, 76.3; H, 9.7. Found: C, 76.3; H, 9.6. This reaction is analogous to the conversion of nologenin (XXIV) to pennogenin (XXV). Mild oxidation of XXVII followed by hydrolysis of the intermediate ester (XXVIII) gives 5,lG pregnadien-X(/3)-01-20-one(XXIX), isolated as its acetate, m. p. and mixed rn. p., 1'76'. Anul. Calcd. for CS8H32O3:C, 77.4; H, 9.1. Found: C , 77.1; H, 9.0. We propose structure XXVII for kappogenin.
New Sources for Known Sterols Sarsasapogenin.-We have described above the isolations and structures of nine new steroidal sapogenins, isolated from either Yuccus or Agmes. We have found that these plants are particularly good sources for other steroidal sapogenins. Thus, we have found twenty-six new sources for sarsasapogenin (XXX). Previous to this work only three sources were known. 1.
June, 194.3
ISOLATION AND STRUCTURES OF SOMEN E W STEROIDAL SAPOGENINS Yield . per kg. i h y )
Plant
Location
Agave allentua (Baker) Agave roealiana (Baker) L,echupuilZa espadilla ntoiado Maguey d e la pena or estvella Smilax lancelata (L) Smilax rotuizdifolia YUCCG angustissima Y . arieonica (McKei.) Y . baccata (Torr.) Y . baleyi (Woot-Standl.) Y . conpnis (McKel.) Y , decipiens (Trel.) Y . endlichiana (Trel.1 Y . eleta (Engelm.) Y . d a t a cultivated young Y . glauca (Nutt.) Y . harrimanii (Trel.) Y . jalicensis (Tre1.j Y . rigida Y . schidigera (Roezl.) Y . scholtii (Engelm.) fruit Y . lhoinbeii (McKel.) Y . lorveyi (Shafer) Y . tveculeana succ. Y . tveculeana can. (Hook.1 Y . walida
St. Louis, Mo. Puebla, Mexico Tehuacan, Mexico Zitacuaro, Mexico South Carolina South Carolina S. W. Utah Nogales, Ariz. LOSCruces, N. Mex. Meteor Crater, Ariz. Douglas, Ariz. Zitacuaro, Mexico Parras, Mexico Tucson, Ariz. Tucson, Ariz. Northern Texas Zion Park, Utah La Primanera, Mexico Western Mexico Palm Springs, Calif. Sonoita, Ariz. Tucson, Ariz. Big Bend, Texas Sonora, Texas , rxico Tamazuchane,M Western Mexico
0.2 0.3 0.6 0.4 5.0 2.8 1.5 0.1 0.8 0.9 1.0 0.1 1.1 10.0 0.6 5.0 1.7 0.4 1.0 2.0 5.0 0.7 2.0 2.7 2 0 3 .0
The identity of the sarsasapogenin was established by analysis of the genin and its acetate along with mixed melting point determinations on both. In all cases the melting points, mixed melting points and analyses of the genin and its acetate were within the following ranges: genin, m. p. and mixed m. p., 199-202'. Anal. Calcd. for C2iH4103: C, 77.8; H , 10.6. Found: C, 77.8 * 0.3; H, 10.6 * 0.3; acetate, m. p. and mixed m. p. 138-142' or 1%-129'. Anal. Calcd. for C2YH4604: C, 73.9; H, 10.1. Found: C , '76.1 * 0.3; H, 10.2 * 0.3. 2. Smilagenin.-Previ~usly,~ there has been reported only one source for smilagenin (XIV), namely, Smilax ornata (Hooker). We have found twenty-eight new sources, listed in the table. Plant
Agave funkiana (Koch-Bouche) A.funkiana var. (Koch-Bouche) A . heterocantha (Zucc.) A , lechuguilla (Torr.) 4.lophantha Dvacena australis Yucca aloifolia (L.) Y . aloifolia var. (Naudin) Y . arkansana (Trel.) Y . australis (Engelm.) Y . brevifolia (Engelm.) New Yucca Y . elephantipes (Regel) Y . lilifera (Chaband) Y . fEaccida (Haw.) Y . gloriosa (L.) Y . jalicensis (Trel.) 1'. louisianensis (Trel.) 1 . vecuvvifolia (Salish ) 1'. veverchoni (Trel
Yield, g. per kg. (dry) Victoria, Mexico 0.8 0.9 Pachucd, Mexico Laredo, Texas 1.3 Big Bend, Texas 5.0 San Antonio, Texas 7.5 3.0 Laredo, Texas 3.0 Galveston, Texas Beaumont, Texas 0.8 5.6 San Antonio, Texas Monterey, Mexico 0.2 0.1 San Bernardino, Calif. 0.1 Big Bend, Texas 0.6 Cordoba, Mexico 0.5 Tehuacan, Mexico 8.0 Tryon, N. C. Gainesville, Florida 1.7 Western Mexico 1 .0 I 11 Heaumont, Texas I 1 I Heaiimont , Texas n ii Sonora, Texas Location
( 7 ) Askew, Farmer and Koa, .I C h r i v \ o r , 1399 (193hj
Y . rastrato (Engelm.) Big Bend, Texas Y . rupicola (Scheele) S. Central Texas Y . scholtii (Engelm.) flowers Sonoita, Ariz. Y . fenuistyla (Trel.) Beaumont, Texas Y . lveleaseuna (MacBride) Beaumont, Texas Samuela carnerosona (Trel.) Big Bend, Texas Samucla faxaniana (Trel.) Big Bend, Texas Zygadenus glaberrimus (Michx.) Myrtle Beach, S. C. Zygadenus nutallii (Gray) Central Texas
1207 1.6 1.1 1.2 2 0 0.5 0.5 0.7 2.5 2.0
Smilagenin (XIV), which may be prepared by the action of ethanolic hydrochloric acid on sarsasapogenin (XXX), differs in structure from the latter only in the ~ide-chain.~The identity of the smilagenin was established by analysis of the genin and its acetate along with mixed melting point determinations on both. Smilagenin acetate resembles sarsasapogenin acetate in showing polymorphic forms. These melt a t 110, 130 and 152'. In all cases the melting points, mixed melting points and analyses of the genin and its acetate were within the following range : genin, m. p. and mixed m. p., 183-185'. Anal. Calcd. for CZ7Hl4O3:C, 7 T . S ; H, 10.6. Found: C, 77.8 * 0.3; H, 10.6 * 0.3. ; acetate, m. p. and mixed rn. p., 10ci-llOo, 126-130' and 149-152'. Anal. Calcd. for C2$H4604: C, 93.9; H, 10.1. Found: C, 75.9 * 0.3; H, 10.1 * 0.3. 3. Gitogenin-We have found sixteen new sources of gitogenin (VI), ten new sources of tigogenin (11) and two new sources for chlorogenin (XXXI). Previous to this work only three sources were known for each of these sapogenins. The gitogenin plants are : Plant Agave gracilipss (Trel.) A . huachucensis (Baker) A . lechuguilla (Torr.) A . mescal (Koch) A . schotlii (Engelm.) A . stricta glauca A . slvicta nana A . slricto rosea Manfreda giganlea gay. Manfreda uirginica (L.) Yucca jilamenlosa (L.) Y . schottii (Engelm.) Y . whipplei coespiloso (Torr.) Y . whipplei intermedia Y . whipplei percursa
Yield, g. per kg. Location (dry) El Capitan Mt., Texas 0.3 Fort Huachuca, Ariz. 0.3 Big Bend, Texas 0.6 Mexico City, Mexico 2.0 Tucson, Ariz. 11.0 Parras, Mexico 1.5 Saltillo, Mexico 1.1 Laredo, Texas 1.1 Laredo, Texas 2.1 South Carolina 10.0 South Carolina 11.0 Santa Catalina Mts., Ariz. 1.0 Palmdale, Calif. 3.0 Santa Monica Mt., Calif. 0.3 Santa Barbara, Calif. 2.0
The identity of the gitogenin was established by analysis of the genin and its acetate along with mixed melting point determinations on both. In each case the melting point of gitogenin was within the range 264-268' and the analyses never varied more than three-tenths from the calculated carbon and hydrogen values. The meltiiig point of gitogenin &acetate was within the raiigc 238-242" and the analvses nevrr Lrarietl i i i o r c tliaii
120s
X\RKEK, ~VACNEK,CISHAFER,WITTBECKEK, GOLDSMITH AND Kuor
two-tenths from the calculated carbon and hydrogen values. 4. Tigogenin. Plant Agave lophanlha A . slricta purpurea A . scholfii (Engelm.) Allium lvicocum Hesperaloe paruijora (Torr.) Maguey Canon del Abra Solanum dulcamara (L.) Yucca whigplei infermedia (Torr.) Y . whipplei parishii Y . whipplei typica
Yield, g. per kg. Location (dry) 1.5 Monterey, Mexico Laredo, Tex. 2.5 Tucson, Ariz. 1.4 Bradford, Penna. 0.1 San Antonio, Tex. I .0 Victoria, Mexico 0 4 Commercial Product 0,l Santa Monica M t . , Calif. 1 , 0 Canyon Pass, Calif. 4.4 San Bernardino, Calif. 8.8
D. dugessi D.galeotliana D. yi.andifdolia I?. hirsuta D. hirsuticairlis D . jaliscana D.l o b d o D. macroslachya I ) . mcxicaiia D.inililaris D . minima D. mulliizervis D . plolycalpata D. plumifera D . firinglei 17. remotiRora D . subtomentosa D . lestudinaria D . ulinei D. urceolafa Nolina erumpens Nolina greciii Svnilacena slellalu 'Z'dlium slylosu MI T . sessile califovni
Guadalajara, Mexico Oaxaca, Mexico Cuernavaca, Mexico Iguala, Mexico Cuernavaca, Mexico Guadalajara, Mexico Cuernavaca, Mexico Vera Cruz, Mexico Central Mexico Uruapan, Mexico Patzcuaro, Mexico Patzcuaro, Mexico Iguala, Mexico Jalapa, Mexico Guadalajara, Mexico Guadalajara, Mexico Cuernavaca, Mexico Guadalupe, Mexico Cuernavaca, Mexico Cuernavaca, Mexico Alpine, Texas N. New Mexico Westerly, R. I. Greenville, S. C .
lrOl.
GT,
2 0
0 2
u . .1
4.0 1.3
0.2
0.L
0.2
0.1
0.2 0.4 0.2 0.2
0.1
0.7
0.6
3.0 1.5 3.1 5.0 2.9 3 9 3.3 2.7 2.7 3.0 4.1
0.1 0.1 0.1
3.5 3.7 4.4 5.0 4.0 4.6 0.5 4 . (1 L O 0.1
0.3
0.3 0.2 In each case the melting point of tigogenin was 0.1 0.2 within the range 205-208O and the analyses of the genins and their acetates were within the above limits. The melting point of tigogenin acetate was within the range 200-204'. Ukiah, Calif 0.1 It is noteworthy that the subspecies of YLLCLU C U whipplei (Torr.) aiid Agave strictu contain different The identities of the above were established by steroidal sapogenins. Thus, Yucca whipplei the analyses of the genins and their acetates along (Torr.) subsp. caespifosa and subsp. percursa with mixed melting point determinations on both. contain gitogenin, subsp. typica and subsp. In each case the melting point of diosgenin, purishii contain tigogenin ; aiid subsp. inter- yaniogenin and kryptogenin was within the ranges media contains gitogenin and tigogenin. Fur- 200-202', 199-201' and 187-189', respectively, thermore, three of the four subspecies of Agazie and the analyses never varied more than threestrictu contain gitogenin, the other contains tigo- tenths from the calculated carbon and hydrogen genin. values. The melting points of diosgenin acetate, 5. Chlorogenin. yamogenin acetate and kryptogenin diacetate Yield were within the ranges 200-202°, 180-182' and g per kg 1W--1Zio, respectively, and the analyses never Plant Location (dry) varied more than three-tenths from the calculated Algutle utahenszs (Engelni St George, Utah 1 ;7 Jfaguey cacuya OriLaba, hlexico 0 1 carbon and hydrogen values. 7. Sitosterol.-In the course of our investigaIn each case the melting point of chlorogenin tion of plants for steroidal sapogenins, we have was within the range 276-278", and the analyses found that the following plants, listed with their of the genins and their acetates were within the geographical location, contained sitosterol as above limits. The melting point of chlorogenin their principal steroidal constituent. No steroidal diacetate was within the range 153-155'. sapogenin could be isolated, except in the case of 6. Diosgenin.-We reported previously the Yucca arizonica and Trillium sessile from which isolation and structures of yamogenin (XX) and we obtained sarsasapogenin and diosgenin, re kryptogenin (XXVI) from Dioscorea testudinuria spectively . and Beth root,Bb respectively. Both substances 11%
are structurally related to diosgenin, and i t is of interest that all three occur in many of the Dioscoreus. With the present and the fourteen previously reported sources, diosgenin is the commonest steroidal sapogenin, having been isolated from forty-three plants. Plant Dsoscorea bzclbtferera D. caprllarrs D composifa I ) cvohorou hi:
Yield, g. per kg. (dry) KryPLocation Diosg. Yamog. tog. Puebla, Mexico 4 5 0 2 0 3 Oasaea, Mexico 2 2 Oaxaca, Mexico 3 0 l ~ i i a l n('.ifion, M r \ i r ~ ) 2 0 I1 I I) 1
Yield, Flant
Aguve itzellzjlua (Trel.) A palmarts (Trel ) A maryllzs Neciattn rosea Hechtia srar70su Lily of the Valley Maguey mescal neul -If mescal bermgo df mescal chuto m e s c d muma largci 1I
/Pq/(111/ / i l l ( / J
Location
P perkg
(dry)
La Union, Mexico Amatitan, Mexico Apoka, Florida Tehuacan, Mexico Hot Springs, Texas Commercial Product Arenal, Mexico Amatitan, Mexico Amatitan, Mexico brenal, Mexico
0 1
I'a~iaqiiilla,hfeiico
0 1
0 3
0 1 0 2 0 1
0 1 0 2 0 2 0 2 0 4
June, 1943
WATER-SOLUBLE DERIVATIVES OF MENADIONE Yield, g. per kg.
Plant
Location
Nolina bigelovi (Torr.) N . macrocarpo (Wats.) N . paraoiflora (H. B. K.) N . texana (Wats.’, Trillium sessile californicum Yucca arizonica (McKel.) Y . thompsoniana
(dry)
Goldroad, Ark. Central Arizona Tehuacan, Mexico Central Texas Ukiah, Calif. Sonoita, Ark. Big Bend, Texas
0.4 0.1 0.1 0.2 0.1 1.4 0.1
The identification of sitosterol was established by the analyses of the sterol and its acetate along with mixed melting point determinations on both. Sitosterol melted 133-136’ and its acetate melted 123-126’. In all cases the analyses never varied more than three-tenths from the calculated carbon and hydrogen values. Of particular interest are those Magueys used for the manufacture of an intoxicating liquor
[CONTRIBUTION FROM
THE
1209
known as tequila. These include M . mescal azul, M . mescal bermejo, M . mescal chato and M . mescal mano hrga. In the course of time the natives have singled out these species as the best sources for its manufacture. Although many Magueys contain steroidal sapogenins, all of these above contain sitosterol.
Summary 1. Some three hundred additional plants have been investigated for steroidal sapogenins. 2. From sixty-six plants thirteen new sapogenins have been isolated. Structures are suggested. 3. From one hundred and sixty-two plants, eight previously known sapogenins and sitosterol have been isolated. STATECOLLEGE, PENNA.
RECEIVED APRIL28, 1943
CHEMICAL LABORATORY, AMERICAN MEDICAL ASSOCIATION]
Water-Soluble Derivatives of Menadione’ BY AMELR. ME NOT TI"^) The antihemorrhagic activity of sulfonated derivatives of 2-methylnaphthalene has been the subject of publications by Moore2 and Baker, Davies, McElroy and Carlson.a The results of the present investigation provide further proof that (a) the primary product formed from the interaction of sodium bisulfite with menadione (2-methyl- 1,4-naphthoquinone) is a monobisulfite addition product, and (b) that this addition product rearranges under the influence of heat to form the sulfonated derivative, 2-methyl-1,4naphthohydroquinone-3-sulfonate. These two products have been characterized, and methods are herein described for their identification in pharmaceutical preparations. Menadione, when shaken with a saturated aqueous sodium bisulfite solution at room or slightly elevated temperature, yielded a clear, yellow solution which, on cooling t o Oo, deposited a white, crystalline, trihydrated salt having the (1) The Council on Pharmacy and Chemistry of the American Medical Association has accepted the name “Menadione” a s a non-proprietary designation for Z-methpl-l,4-naphthoquinone” (J. A . M . A . , 116, 1054 (1941)),( l a ) Present address, Cheplin Biological Laboratories, Syracuse, N. Y., and the name “Menadione Bisulfite as a non-proprietary designation for the water soluble monosodium bisnlfite addition product of menadione” described in this paper ( J . A . M .A , , 131, 839 (1943)). (2) M. B. Moore, THISJOURNAL, 68, 2049 (1941). (3) B. R. Baker, T. H. Davies. L. M u E l r o ~Bad G. W. Corlmn, ibid , 64, 1096 (1942).
characteristics of a bisulfite addition product. If the solutions were heated a t 90-100’, the quantity of bisulfite addition product obtained on cooling t o 0’ decreased with increase in heating time. After heating for twenty to thirty hours, no crystalline organic salt precipitated on cooling or on the addition of organic solvents. However, if saturated potassium chloride solution was added, the cooled solution deposited crystals of a potassium salt which exhibited properties of an organic sulfonate. The optical properties of the sulfonate and its behavior toward dilute alkalies, o-phenanthroline-ferrous complex solution and ethyl cyanoacetate, differed radically from the menadione bisulfite addition product. The sulfonate could be detected in the mother liquid, by precipitation with o-phenanthroline-ferrous complex, when the bisulfite addition product was made from warmed solutions. The greater solubility of the sodium salt of the sulfonate in contrast with that of the addition product made possible a complete separation on cooling the reaction mixture. The crystallized, purified menadione-sodium bisulfite addition product was converted to 2-methyl-l,4-naphthohydroquinone3-sulfonate in 20-30y0 yield by solution in water and heating a t 100’ for twenty-four hours in a closed vessel. No bisulfite addition product
