Extraction And Separation Of Fungistatic Compositions From Kochiae Scopariae And Studies On Its Antifungal Activity

Kochiae Scopariae was a traditional Chinese medicine used as anti-bacterial, anti-inflammatory, anti-viral, anti-allergic, anti-gastric mucosal injury, lowering blood sugar, enhancing immunity and so on. Kochia scoparia compound preparation was effective in treating skin disease, urinary tract infections, acute mastitis and other diseases.The anti-phytopathogenic fungi activity of Kochiae Scopariae was studied to use Colletotrichum capsici as tested strains, in order to develop new botanical fungicide, and the effective components were separated. The results were as follows:1. Water extract of Kochia Scoparia had an excellent antifungal activity on plant pathogenic fungi, and the inhibition rate was all more than 50%. At 106 mg/L, extract had the best inhibitory effect on Colletotrichum capsici with the rate of 99.96% and EC50 value of 2.28×104 mg/L. The inhibitory effect was better than the control. At 105 mg/L, the spore germination rate was 95.21% with EC50 value of 4.04×103 mg/L.2. The optimal extraction condition of fungistatic active ingredients from Kochia Scoparia were obtained as follows: extraction time 48 h, extraction temperature 25℃, solid-liquid ratio 1:20. In this condition, inhibition rate of extract reached 97.18%.3. Separated by macroporous resin, 20% ethanol eluate showed the highest inhibition rate against Colletotrichum capsici reached 100% with the EC50 value of 5.46×103 mg/L.4. The chemical constituents of the active site had been tested systematically by tube-test. It contained flavonoids, terpenes and alkaloids.5. The test on stability of active site seperated by macroporous resin showed that: above the temperature of 60℃, the extract of the active site had not stability of the activity; and the ultraviolet irradiation had no significant effect on the inhibition of the active ingredient; and the antimicrobial active ingredients had great stability under acidic conditions; Antifungal activity was increased by adding metal ion, particularly the Na+.6. The high-speed countercurrent chromatography and silica gel column chromatography was used for the further purification, the active ingredients were isolated and identified by nuclear magnetic resonance spectroscopy. The result showed that one of the active constituent was oleanolic acid. The compound had great antifungal activity with the EC50 value of 1.12 × 103mg/L.