| Allii macrostemonis bulbus is dried bulbus of Allium macrostemon Bge.or Allium chinense G.Don,which are plants of liliaceous family.It has many pharmacological activity,such as anti-cancer,anti-oxidation,lipid-lowering,antispasmodic,bacteriostasis and inhibition of atherosclerosis.It can be used in treatment of coronary heart disease,angina pectoris,bronchial asthma,hyperlipidemia and myocardial ischemia etc.In this paper,18 compounds(marked as XB-1~XB-18)had been obtained from 75% ethanol extract of Allii macrostemonis bulbus and 15 of them were identified by means of various spectroscopy methods combining with the physical and chemical properties,including 2 new compounds and 1 compound obtained from this medical material for the first time.The two new compounds are(25S)-26-O-?-D-glucopyranosyl-5?-furostane-2?,3?,22,26-tetraol-3-O-?-D-glucopyranosyl-(1?2)-[?-D-glucopyranosyl-(1?3)]-?-Dglucopyranosyl(1?4)-?-D-galactopyranoside(compound XB-15)and 25(27)-ene-26-O-?-D-glucopyranosyl-5?-furostane-3?,22,26-triol-3-O-?-D-glucopyra-nosyl-(1?2)-[?-D-glucopyranosyl-(1?3)]-?-D-glucopyranosyl(1?4)-?-D-galactopyranoside(compound XB-16).The first isolated compounds is 25(27)-ene-26-O-?-D-glucopyranosyl-5?-furostane-2?,3?,22,26-tetraol-3-O-?-D-glucopyranosyl-(1?2)-[?-D-glucopyranosyl-(1?3)]-?-D-glucopyranosyl(1?4)-?-D-galactopyranos-ide(compound XB-12).On the basis of separation and identification of chemical constituents,the experiment established a SPE-ELSD-HPLC method to determine the contents of macrostemonoside A(compound XB-1)and(25R)-5?-spirostane-3?-3-O-[?-D-glucopyranosyl-(1?2)-?-D-galactopyranoside](compound XB-4).After investigation to extraction solvent,extraction method,extraction volume and solvent time which were factors affecting the extraction rate,we have established the optimum extraction method,and then try to study methodology.The results show that the two compounds had a good linear relationship with the ranges of 0.40~20.00?g and 0.20 ~10.00?g respectively.Both compounds had a good performance in precision,stability,repeatability and recovery with the RSDs less than 2%.Then two batches of samples of Allii macrostemonis bulbus were determined,the results showed that the average contents of compound XB-1 and compound XB-4 were 0.21 mg·g-1 and 0.07 mg·g-1,respectively.In this paper,the activity experiment of proliferation inhibition on cell of Hep G2,melanoma A375,breast cancer MCF-7 and lung cancer A549 have been peformanced by CCK-8 method for the new furostan saponins(compounds XB-15,25S-type)and its isomers(compound XB-14,25R-type).The results showed that each 25 R/S isomers,compounds XB-14 and XB-15,had weaker inhibition on the cells of lung cancer A549,A375 melanoma cells and breast cancer MCF-7 cell,had certain inhibitory effect on Hep G2 cells,but the inhibitory effect of R configuration of compound XB-14 is stronger than XB-15.It illustrated that the same compounds have different proliferation inhibition on different tumor cells,meantime,the difference of R/S structure could also lead to the difference inhibition effects on tumor proliferation.In summary,this paper had gained and identified 15 compounds from 75% ethanol extract of Allii macrostemonis bulbus including 2 new compounds and 1 compound obtained from this medical material for the first time.The contents determination methods of two spirostanol saponins of Allii macrostemonis bulbus using SPE-ELSD-HPLC were established.The inhibitory effects of R/S iosmers compounds XB-14 and XB-15 on the proliferation of tumor cells were studied.The results of this paper provided new scientific basis for further research on the basis of pharmacodynamic substance,perfection of quality standards of Allii macrostemonis bulbus and application of steroidal saponins from this herb in anti-tumor aspect. |
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