~ I A D HJAIS, L - -1.K. R.ITHORE*and 5.C. J I I S D e p a r t m e n t qf Botany, L-nizersity of Rqjiisthan, Jilipur, Indzo t h e characteristic colors developed 118). Five spots were labelled (-1 t o E ) in order of decreasing R f value. I n a parallel chromatogram spots C, and E in the flowers and roots corresponded t o sitosterol, cho1es;erol and diosgenin respectiwly, but in t h e aerial p a r t s spots D and E coincided with sitosterol and cholesterol. Bands C , D , and E were separated by preparative tlc on air-dried silica gel G layers (wet thickness 500 mp'i by using two-fold development in hexane-ethyl acetate t3:11 and spraying with anisaldehyde reagent. Bands coinciding m-ith sitosterol, cholesterol, and diosgenin were marked, scraped from unsprayed plates, eluted with chloroform, tested by tlc for their purity, eluted, and later crystallized (19). Spots d and B in t h e flow-ers and roots, and spots -1,B, and C in t h e aerial p a r t s could not be identified due t o insrifficient quantit?-. F o r the isolated dioseenin we determined: mp 203-201", mnip undepressed, acetate nip 191-195', :a:25D-129', and t h e i r spectrum (characteristic peaks a t 984, 922, 901 and 806 em-'). Sitosterol and cholesterol were identified on t h e basis of t h e LieherniannBurchard t e s t , tlc in various solvents, color reactions, mp, and i r spectral studies. Diosgenin was estimated quantitative1)- in t h e isolates of flowers and roots.
The occurrence of dioagenin has been reported in various Solaiiiim species (1-14). but it has not been reported from s. jasminoities Past. The presence of solasonine. solamargine, and the hydrolytic product solasodine ha.; been investigated in different plant parts of this species (15IG). In continuation of earlier findings. n-e report here the isolation and identification of steroids from flower roots: and aerial parts of S. jasnii-
n,
iioides.
EXPERINESThL PL.LST~IITERI.~L.--.S.jiisniinoides P a s t . plants were collected during October 1 9 i i a t the Botanical Garden of Maharaja's College, Jaipur: India. -1voucher specimen of this collection has been deposited with t h e herbarium of t h e Botany Department, Cniversit)- of Rajasthan, Jaipur, India.
ESTR-ICTIOSOF STEROIDS.-Dried and powdered flom-ers, roots, and aerial p a r t s without flowers 110 g each) of S j m n i i n o i d e s were separately defatted in succession with 100 ml of petroleum e t h e r (24 h r ) and benzene 116 h r ) . T h e defatted plant samples, obtained after filtration, were air-dried, repon-dered and separate1)- refluxed for 4 h r with 20 ml g of 5s HC1 in 7 0 7 ethanol. Each hydrolyzate solution (approximately 200 m l ) obtained after filtration was repeatedly estracted with e t h y l acetate i l i ) for complete extraction of sapogenins. 1-arioils fractions were pooled, dried in i'ilcuo and taken up in chloroform for further analyses.
ISOLITIOS .LSD IDESTIFIC.ITIOS
COLORIMETRIC ESTIXITIOS OF DIOSGESIS .Diosgenin was quantitatively estimated by t h e method of Sanchez et ill. (20) after tlc on silica gel G with n-hesane-ethyl acetate (3:l). -1bsorbances measured with a spectrophotometer (Carl Zeiss, ,Jena D D R , T-SU-2 P) set a t 405 nni against a blank m-ere referred t o a calibration curve which followed t h e Beer's law. T h e concentrations of t h e two extracts (flowers and roots) were determined by taking t h e mean values of 15 replicates.
OF S T E R O I D S .
RESULTS Diosgenin and tn-o sterols (sitosterol and cholesterol) not knon-n to occur in the plant species under investigation have been isolated and identified both from flon-ers and roots. but the other aerial parte n-ere found t o lack diosgenin. Higher diosgenin content \\-as pre.sent in roots (0.33% dry weight ) as compared to flon-ere (0.1SX ). Since the fruit
-Each crude steroid extract was examined b>- two-dimensional tlc on air-dried silica gel G iwet thickness 250 nip) using dichloromethane-methanol-formamide(93:G:l'i i n t h e first direction and cyclohesane-ethyl acetate-water (Ci00:400:1) in t h e second direction 118). T h e steroidal compounds were located by spraying with 5 0 5 sulfuric acid and, also separate1)- TT-ith anisaldehyde reagent 10.5ml of anisaldehyde in 50 ml of glacial acetic acid and 1 nil of concentrated sulfuric acid) and then heating at 100" until *Ilepartnient of Horticnltiire, Univer.sit>of Udaipiir, Udaipiir. India.
421
422
JOCRNAL OF SATURAL PRODUCTS
setting is very 10~1-in the plant under cultivation, the occurrence of diosgenin in the flowers is noteworthy. ACKNOWLEDGMESTS We wish t o thank the University Grants Commission, New Delhi, India, for financial assistance granted to one of u s (SCJ) and t o thank Dr. (hliss) Pushpa Khanna for valuable suggestions. Recekled 98 4 u g z i s t 1978. 1.
2. 3.
4. 5.
6.
LITERATURE CITED Y. SATOand H. G. LATHAM, JR., J . d n i e r . Chenz. Soc., 73, 6067 (1953). 0 . S. hl.AD.AEr.4 and L . F. STEP.ASOVA, X e d . P r o m . SSSR., 19, 49 (1965). 6 . ~ ~ O O K H E R J Eand A R . hIATHETT-, Jour. I n d i a n Ciiem. Soc., 46, 749 (1969). I . P. 1-ARSHSEY and S . K. DCBE,Jour. I n d i a n Ciiem. Soc., 47, 717 (1970). E. GUERREIRO, 0. S. GIORDASO,J . KAVKA and 8.T. DJL4RCISGERLO,A n . QuZn7., 67, 789 (1971). A. U R Z U Aand B. K . CASSELS, Piiytochemistry, 11, 3548 (1972).
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17. 18. 19. 20.
[VOL.
42, so. 4
0 . S. GIORD.-, J . K.4vK.1, J . C. GIASELLOand A . T. D’ARCASGERLO, An. Asoc. Quim. A r g e n t . , 61, 47 (1973). A. -4. SALEH,P l a n t a M e d . , 26, 40 (1974). W. DOEPKE,U. HESSand G. PADROS, P h a r m a z i e , 31, 133 (19i6). W . DOEPKE,I. L. MOLAand U. HESS, P h a r m a e i e , 31, 666 (1976). 1%‘. DOEPKE,v. JIMENEZ and u. HESS, P h a r m a e i e , 31, 488 (1976). P. KHISSAand A. K . RATHORE, Indian J. Exptl. Biol., 13, 808 (1977). R . CARLE,A. W. ALFERMASSand E. REISHARD, P l a n f a M e d . , 32, 195 (1977). R.SEGAL,I. hkLO-GOLDzxEIG and D . T. ZAITSCHEK,Lloydia, 40, 604 (1977). 6. C. JAISand G . L. S H m x i , P l a n f a M e d . , 31, 212 (1977). S. C. J a m and G. L. SH.ARM.~, Pianta X e d . , 32, 233 (19i7). Y.TOMITA, A. UOMORIand H. MIXATO, Phytochemistry, 9, 11 (1970). R . D. BENXETT and E. H E F T M A X S , J. Chromntogr., 9, 353 (1962). S. C. JAIS, L l o y d i a , 39, 244 (1976). G . L . SANCHEZ, J . C. MEDISAACEX-ADO and R . R . SOTO, A n a l y s t , 97, 973 (1972).
