Antibacterial Ant Ioxidant Effect Of Extract And Purified Of Chlorogenic Acid And Astilbin From Smilax China L.

Smilax china L.(SCL) is a medicinal plants with several functions, such as antibacterial, antitumor, hypoglycemic, antioxidation, anti-inflammatory. Astilbin and chlorogenic acid(CGA)are the main functional compounds in SCL. In the present study,the antioxidant and antibacterial activities of the ethonal extract(ET), petroleum ether phase(PEP), chloroform phase(CP), ethyl acetate phase(EAP), butanol phase(BP) and water phase(WP) from SCL is investigated, then the CGA and astilbin is extraced and purified from the WP and the EAP. The results were showed as follow:(1) The antibacterial activities of the extracts were tested by the filter paper method,the minimum antibacterial concentration and half antibacterial concentration. The results showed that the extractions of SCL had antibacterial effect, and the antibacterial activities of the extractions were shown as followed: EAP > BP > WP > ET > PEP > CP. The 80mg/mL EAP has the best antibacterial effect with the inhibition zone 13.6±0.12, 13.2±0.10,12.9±0.07, 12.5±0.01(mm) for staphylococcus aureus, bacillus subtilis, salmonella and Escherichia.coli, respectively.(2) Using BHT as a positive control, the antioxidant activities of the extracts were tested and compared by ·DPPH, ·OH, ·O2-, ·ABTS+, and the FRAP with the concentrations of 0.2-1.2 mg/m L. The results showed that all extractions had antioxidant effects, and had concentration-response relationship. The EAP has the strongest antioxidant activity. The abilities of antioxidation were shown as followed: EAP > BP > WP > ET > PEP > CP. The ability to clean ·OH of EAP is weaker than that of BHT, but the ability to clean·DPPH, ·OH, ·O2-, ·ABTS+ and total antioxidative of EAP are stronger than BHT.The scavenging capacity to ·OH, ·DPPH, ·O2- and ·ABTS+ were 58.73%, 66.46%, 73.17%and 95.84%, respectively, FRAP value was 186.25±0.21.(3) Based on the response surface analysis with the Design-Expert software program,the extraction conditions of the microwave for CGA in SCL were optimized. Optimum process conditions was obtained using one factor test and response surface test: volume fraction of ethanol 50%, microwave power 300 w, ratio of liquid to solid 30:1(mL/g),microwave time 87 s, temperature 48 oC. The theoretical yield of chlorogenic acid was9.4565 mg/g and practical yield was 9.3972 mg/g, relative error was 0.627%.(4)The S-8 was selected as optimal resin from seven kinds of resins. The optimum adsorption-desorption conditions were as follows: the pH was 2.0, the Weight of resin was1.0 g/30 mL, the concentration of sample was 80 mg/mL, the flow rate of sample was1.0mL/min, volume fraction of ethanol in eluent was 50%, the speed of elution was 1.0mL/min, the volume of elution was 5 BV. The CGA was eluted by 50% ethanol with purityof 76.47%. Then the further purifing by silicone column chromatography, the spots on the same location were collected by thin layer chromatography(TLC). The CGA product was further identified by HPLC, 1H NMR and 13 C NMR.(5)Astilbin in ethyl acetate layer was absorbed by the polyamide and the optimum adsorption-desorption conditions were shown as followed: the concentration of sample was80 mg/mL, the flow rate of sample was 1 m L/min, volume fraction of ethanol in eluent was50%, the speed of elution was 1.0 mL/min. Then the further purifing by silicone column chromatography, the spots on the same location were collected by TLC. The Astilbin product was further identified by HPLC, 1H NMR and 13 C NMR.Through this paper found that each extract of SCL have antibacterial and antioxidant effect, EAP has the strongest antioxidant and antibacterial activity. The abilities of antioxidant and antibacterial were shown as followed: EAP > BP > WP > ET > PEP > CP.The CGA is purified from the ethyl acetate layer, the astilbin is purified from the water layer, and their purity is greater than 90%.