Study On Chemical Constituents And Antibacterial Activity Of Ethyl Acetate Extract From Angelica Sinensis

Angelica sinensis(Oliv.) Diels mainly distributes in Gansu, Yunnan, Sichuan of China. Its dry root is a famous traditional Chinese medicine and exhibt a warm character, which tastes a bit sweet and spicy. It has many pharmacological activities, including treating symptoms of blood deficiency, regulating menstruation, relieving pain, lubricating intestine and so on. Many researches including, species, morphological characteristics, chemical compositions and pharmacological activity of medicinal plants belonging to Angelica.L was summarized in this paper. The chemical compositions of ethylacetate part from Radix angelicae sinensis were also separated and purified with the guidance of antibacterial activity.(1) Antimicrobial activities of four polar fractions from Radix angelicae sinensis by broth microdilution method were measured. The test strains used in the experiment was Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, Pseudomonas aeruginosa, Bacillus licheniformis, Klebsiella pneumoniae. The result showed that petroleum ether and ethyl acetate parts exhibited stronger antibacterial activity. MIC of petroleum ether part against six test bacteria were between 62.5~125?g/mL, its MBC were over 1000?g/mL against both S.agalactiae and P.aeruginosa while its MBC were between 500~1000?g/mL against other four test bacteria. MIC of ethyl acetate part against six test bacteria were all 125?g/mL, its MBC were over 1000?g/mL against both S.agalactiae and P.aeruginosa while its MBC were between 250~500?g/mL against other four test bacteria. MIC of n-butyl alcohol extract against S.agalactiae was 500?g/mL while MIC against other five test bacteria were all over 1000?g/mL, and its MBC were over 1000?g/mL. MIC of water parts was over 1000?g/mL.(2) 12 fractions were obtained by silica gel column chromatography from ethyl acetate extract and their antibacterial activities were measured by broth microdilution method, the result showed that eleven fractions have different antibacterial activities against the six test strains except Fr7, among which Fr5, Fr6, Fr8 and Fr9 have stronger antibacterial activity. MICs of Fr5 and Fr8 against six test bacteria were between 125~250?g/mL, their MBCs were between 250~500?g/mL. MICs of Fr6 and Fr9 against six kinds of test bacteria were between125~500?g/mL, and their MBCs against P.aeruginosa were both over 1000?g/mL while MBCs against other five test bacteria were between 500~1000?g/mL. MICs of Fr1, Fr2, Fr3, Fr4 Fr10, Fr11 and Fr12 against six kinds of test bacteria were between 250~1000?g/mL. All MBCs of Fr1, Fr3 and Fr4 against P.aeruginosa were 1000?g/mL, MBCof Fr3 against K.pneumoniaehas was 1000?g/mL and its MBC against other four test bacteria were over 1000?g/mL. Both MBCs Fr1 and Fr4 were 1000?g/mL against B.licheniformisits while MBCs against other four test bacteria were over 1000?g/mL. MBCs of Fr2, Fr11 and Fr12 were all over 1000?g/mL against the six test bacteria.(3) Ten compounds were separated from ethylacetate extract of Radix Angelicae Sinensis, by using silica gel, Sephadex LH-20 and other columnchromatography. Eight compounds were identified by 1H-NMR, 13C-NMR and ESI-MS spectra and other modern technology, and combined with the physical and chemical properties and related references,their structure were elucidated as 6-methoxycou-marin(1),(E)-2-methoxy-4-(3-carbooxypropenyl)phenol(2), Z-butylidenephthalide(3), E-butylidenephthalide(4), Z-ligustilide(5), Tokinolide B(6), ?-sitosterol(7), Z-6,7-dihydroxy ligustilide(8), Compound(2) was first isolated from this genus. Structural identification and activity determination of remaining monomers compounds will be done later.