Studies On The Active Constituents Of Traditional Chinese Medicine-Trillium Tschonoskii Maxim

The rhizome of Trillium tschonoskii,as a Chinese herb,has been used for the treatment of irregular menstruation,traumatic haemorrhage,bruises and insomnia in clinic.In previous study,T.tschonoskii showed good pharmacological activity and contained several kinds of components such as steroidal saponins,flavonoid glycosides and phenylpropanoid glycosides.The analysis result of the extracts of T.tschonoskii by UPLC-Q-TOF/MS illustrated that a lot of constituents were not reported in literature.A series of separation and analysis methods and strategies were applied to obtain 48 compounds from T.tschonoskii.The separation and analysis methods included macroporous resin column chromatography,silica gel column chromatography,pHPLC and TLC.The separation and analysis strategies included deducing the proportion of organic phase in the mobile phase to improve the resolution,increasing the proportion of organic phase in the mobile phase to improve the resolution,changing the type of organic phase and used the seldom used acetone as the organic phase to improve the resolution,changing the type of the stationary phase and used the positive stationary phase to improve the resolution.Their structures contained 29 new compounds were elucidated by HR-ESI-MS,ORD,1D-NMR and 2D-NMR.The new compounds included 10 new furostanol saponins and 19 new spirostanol saponins,such as 26-O-?-D-glucopyranosyl-?25R?-16,22-dione-furostan-3?,5,6,26-butanol-3-O-?-L-rhamnopyranosyl-?1?4?-O-?-L-rhamn opyranosyl-?1?4?-[O-?-L-rhamnopyranosyl-?1?2?]-O-?-D-glucopyranosi de?1?,?25R?-3?-O-?-L-rhamnopyranosyl-?1?4?-[?-L-rhamnopyranosyl-?1?2?]-?-D-glucopyranosyl-furosta-?^{5?6?,16?17?,20?22?}-triene-26-O-?-D-glucop yranoside?4?,26-O-?-D-glucopyranosyl-?23R,25R?-16,23-cyclocholest-?^5?6?,17?20?-dien-22-one-3,16,26-triol-3-O-?-L-rhamnopyranosyl-?1?4?-O-?-L-rhamnopyranosyl-?1?4?-[O-?-L-rhamnopyranosyl-?1?2?]-O-?-D-glu copyranoside?6?,15-one-18-nor-?23S,24S,25S?-spirost-5,13-diene-1?,3?,23-triol-21,24-acetyl-1-O-?-D-apiofuranosyl-?1?3?-O-?-L-rhamnopyranosyl-?1?2?-[O-?-D-xylopyranosyl-?1?3?]-O-?-L-arabinopyranoside?16?,13-carboxaldehyde-15-one-?23S,24S,25S?-spirost-5-ene-1?,3?,21,23,24-pentan ol-1-O-?-D-apiofuranosyl-?1?3?-O-?-L-rhamnopyranosyl-?1?2?-[O-?-D-xylopyranosyl-?1?3?]-O-?-L-arabinopyranoside?20?and?23S,24S,25S?-spirost-5-ene-1?,3?,13,15,21,23,24-heptanol-1-O-?-D-apiofuranosyl-?1?3?-O-?-L-rhamnopyranosyl-?1?2?-[O-?-D-xylopyranosyl-?1?3?]-O-?-L-ara binopyranoside?27?.Simutaneously,cytotoxic activity of 32 isolated compounds were evaluated against the HepG2 cell lines,and compounds 46,47 and 48 showed significant cytotoxicity with IC₅₀ 4.60?M,1.69?M and1.35?M,respectively.In this study,a series of novel steroidal saponins with different structural features,such as steroidal saponins with carbonyl at C-16 and C-22 position and double bond at?^17?20?position,steroidal saponins with carbonyl at C-15 position,double bond at?^13?14?position and the sugar moiety at C-1 position,were isolated from the rhizomes of T.tschonoskii,which enriched the library of steroidal saponins.It also provided chemical entities for the pharmacological research of T.tschonoskii.Meanwhile,the research lay the foundation for the comprehensive development of T.tschonoskii and the resource replacement of Paridis Rhizoma.