Study On The Chemical Constituents Of Smilax Scobinicaulis And Myrica Rubra And Their Antimicrobial Activity

Smilax scobinicaulis C.H. Wright belongs to family of Smilacaceae. Its rootstalk has been used in folk medicine as antirheumatic disease, move blood stasis and clear toxins, anticonvulsant, curing enteritis and cancer etc. Myrica rubra Sieb et Zucc belongs to family of Myricaceae. Its root can be used as hemostatic and to cure cholecystitis, pericementoclasia and colic etc. However, the chemical constituents and biological activities of the root from Smilax scobinicaulis and Myrica rubra have not been reported. To further exploit and utilize these plant resources and provide a theoretical basis on taxonomy of chemical and clarify the role of pharmacological for Smilax and Myrica plant, this dissertation were carried out a systematic research on specific distribution property of chemical constituents from Smilax Scobinicaulis, the antimicrobial activity of the extract of Smilax scobinicaulis, The separation and structure of the chemical composition from Smilax scobinicanlis and the root of Myrica rubra. The result showed as follows:The distribution property of chemical constituents from Smilax scobinicaulis was studied through the protested systematically and UV method. The result indicated that organs almost have sugar, phenol, digallic acid, aminoacid, protein, alkaloid,saponin, anthoxanthin, enthraqu -inone, coumarin,sterol and lactones etc.except saponin, oumarin and lactones in fruit , fat only in kernel and cardiac glycoside only in pulp. The totalcontent of flavonoids:fibrous root> pulp> root tuber> leaf>stem > rhizome> tendril> kernel. The total content of saponin:leaf > stem > tendril>root tuber> rhizome> fibrous root.Filter-paper diffusion method was adopted in lab to test the antimicrobial activities of seventeen extracts, which concluds twelve solvent extracts and five eluates obtained with macroreticular resin at different ethanol concentrations from the root, stem and leaf of Smilax scobinicaulis. The result revealed that different organs and different extract of the same organ of Smilax scobinicaulis have different inhibiting activity on animalcule, generally inhibit bacteria and almost no effect on fungus. The ethyl acetate extract from stem have strongest inhibitory effect on the tested Staphyloccocus aureus, Bacillus subtilus, B. cereus, E. coli, B. megaterium and Proteus vulgaris. Its MIC respectively is 0.8mg·mL^-1, 0.6mg·mL^-1, 1.2 mg·mL^-1, 1.0mg·mL^-1, 1.0mg·mL^-1 and 1.0mg·mL^-1. The eluate obtained by macroporous resin with ethanol at a concentration of 100% showed a very strong broadband antimicrobial activity. Its MIC on S. aureus, B. subtilus, B. cereus, E. coli, P. vulgaris, B. megaterium, Aspergillus flavus, Aspergillus niger, Trichoderma Koningii, Penicillium sp. and Trichoderma viride respectively is 2.0mg·mL^-1, 1.0mg·mL^-1, 1.2mg·mL^-1, 1.0mg·mL^-1,1.2mg·mL^-1, 1.5mg·mL^-1,0.5mg·mL^-1, 0.5 mg·mL^-1, 0.8mg·mL^-1, 0.8mg·mL^-1and 0.3 mg·mL^-1. Moreover, the eluate obtained by macroporous resin with ethanol at a concentration of 80% showed very strong inhibitory activity on Trichoderma viride and E. coli. Its MIC respectively is 0.2 mg·mL^-1 and 0.5 mg·mL^-1. Else eluate has no inhibitory activity on animalcule.Thirteen steroidal saponins were separated from the root of Smilax scobinicaulis and their structures were identified. The result showed that seven steroidal saponins are new compounds and six steroidal saponins are known compounds by scifinder search. The new compound respectively is (25S)-spirostan-5-ene-3β,27-diol(isonarthogenin)3-O-{β-D- glucopyrnosyl -(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (1),(25S)-spir -ostan-3β,11,27-triol-3-O-{β-D-Glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (3), (25S)- spirostan-3β, 27-diol-3-O-{β-D-Glucopyrnosyl-(1→4)-[α-L- Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (4), (25S)-spirostan-3β,27-diol-3-O-[α-L- Arabinopyranosyl-(1→6)]-β-D-glucopyranoside (5),(25R)-spirostan-6β-ol-3-O-{β-D- glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (7), (25S)- spirostan-5-en-3β-ol-3-O-{β-D-glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (8), (25S)-spirostan-3β,17α,27-triol-3-O-{β-D-glucopyrnosyl-(1→4)-[α-L- Arabinopyranosyl-(1→6)]}-β-D-Glucopyrano-side (9).The known compound respectively is (25S)- spirostan-5-ene- 3β,27-dihydroxy-3-O-[α-L -Arabinopyranosyl-(1→6)]-β-D-glucopyranoside (2), (25R)-5α-spirostan-3β-ol (tigogenin) 3-O-{β-D-glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (6),(25S)-spirostan-5-en-3β,17α,27-triol-3-O-[α-L-Arabinopyranosyl-(1→6)]-β-D-glucopyranoside (10),(25S)-spirostan-5-en-3β,17α,27-triol-3-O-{β-D-glucopyrnosyl-(1→4)-[α-L-arabinopy arabinopyranosyl-(1→6)]}-β-D-Glucopyranosid (11),(25R)-3β-hydroxy-5α-spirostan-6-one(laxogenin)3-O-[α-L-Arabinopyranosyl-(1→6)]-β-D-glucopyranoside (12),laxogenin-3 -O-{β-D-glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside (13). A group of anti-bacterial steroidal saponins from n-butanol extraction of the root from Smilax scobinicaulis were screened. The eluates obtained by macroporous resin with ethanol at a concentration of 50%( R-5), 70%(R-7), 80%(R-8)and 90%(R-9)have stronger inhibitory activity on P. vuigaris; The eluates obtained by macroporous resin with ethanol at a concentration of 60%(R-6), R-8, R-9 and R^-10 have stronger inhibitory activity on E. coli.; R-7, R-8, R-9 and Laxogenin-3-O-[α-L-Arabinopyranosyl-(1→6)]-β-D-glucopyranoside(saponin 1),laxogenin-3-O-{β-D-glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]} -β-D-Glucopyranosid(saponin 2)have stronger inhibitory activity on B. megaterium; R-7,R-8 and R-9 have stronger inhibitory activity on B. cereus and B. subtili.A group of anti-fungi steroidal saponins from n-butanol extraction of the root from Smilax scobinicaulis were also screened. All of steroidal saponins except n-butanol extraction have strong inhibitory activity on T. viride; R-7, R-8, saponin 1 and (25S)-spirostan-3β, 27- diol-6-one -3-O-{β-D-glucopyrnosyl-(1→4)-[α-L-Arabinpyranosyl-(1→6)]}-β-D-Glucopyra -noside(saponin 3)have strong inhibitory activity on A. niger; R-9 , saponin1 3 and (25S)-spirostan-5-en-3β,17α,27-triol-3-O-[α-L-Arabinopyranosyl-(1→6)]-β-D-Glucopyranosi-de(saponin 4)have strong inhibitory activity on T. Koningii; R-7, saponin 1, 2 and 4 have strong inhibitory activity on Penicillium sp.; R-7, saponin 1, 3 and 4 have strong inhibitory activity on A. flavus.Four compounds were separated from ethyl acetate extract of the root from Smilax scobinicaulis and their structure were identified asβ-sistostero(l1) , laxogenin(2), quercetin (3)and daucosterol(4). Compound 2 was isolated from the roots of the plant for the first time.Five compounds were separated from the stem of Smilax scobinicaulis and theirs tructure were identified as resveratol(1), querectin-3-O-β-D-glucoqyranoside(2), luteolin-7-O-β-D-glucoside(3), laxogenin-3-O-[α-L-Arabinopyranosyl-(1→6)]-β-D-Glucopyranoside(4), laxogenin-3-O-{β-D-glucopyrnosyl-(1→4)-[α-L-Arabinopyranosyl-(1→6)]}-β-D-Glucopyranoside(5). Compound 1³ were isolated from the roots of the plant for the first time. Seven compounds were separated from the stem of Smilax scobinicaulis and their structure were identified as myricanol(1), myricanol-5-O-β-D-glucopyranoside(2), 8-hydroxy- myricanone(3), myricanone -5-O-β-D- glucopyranoside(4), porson(5), daucosterol(6),β-sistostero(l7). Compound 1⁵ were biphenyl-type diarylheptanoid. Compound 3 was a new one. Compound 5⁷ were isolated from the roots of the plant for the first time.