Separation,Purification And Characterization Of Active Substance From Gorgonian And Caulerpa Lentillifera

Gogornian and have not been reported applied to industrial production in marine resources, which have been reported that gogornian and caulerpa lentillifera contain corresponding active substances while there has no complete separation and purification system. This paper mainly studied isolating active substance from gogornian and caulerpa lentillifera coupled with HSCCC. Studying its antioxidant activity,providing theoretical and practical basis for further implementing the high-throughput screening of marine active ingredients and the practical application of new antioxidant industry.The study was based on gogornian from south ocean area, the sample was clean by ultrasonic cleaner. Extracting its active substance after being dried assitaned with microwave. Its extraction process was optimized by RSM as follows: liquid-solid ratio was 1:40(g·mL^-1),microwave power was 200 W, extraction temperature was 41℃, ethanol concentration was 82%,under this condition, the actual extraction rate of pseudopterosin from gogornian was 95.21%. Then isolating and purifiding its crude extraction applied with HSCCC. Its solvent system was normal hexane: methyl alcohol:water（2: 1: 1,V/V）.considering the light phase as stationary phase and heavy phase as mobile phase. Collecting the fraction based on the online chromatogram. The molecular mass of fraction I was 480.27908 and molecular Formula was C26H40O8 by mass spectrum. Identifing its structure was pseudopterosin I by NMR. And The molecular mass of fraction II was 423..27372,molecular Formula was C24H39O6 by NMR. Studying its content by quantitative Analysis, it came out the content of pseudopterosin I was 1.21mg·g^-1 and pseudopterosin II was 1.02 mg·g^-1.Caulerpa lentillifera was clean by ultrasonic cleaner. Extracting its active substance after being dried assitaned with microwave. Its extraction process was optimized by RSM as follows: liquid-solid ratio was 1:30(g·mL^-1),microwave power was 200 W, extraction temperature was 45 ℃, ethanol concentration was 85%,extraction time was 20 min,under this condition, the actual extraction rate of pseudopterosin from gogornian was 91.9%.Then isolating and purifiding its crude extraction applied with HSCCC. Its solvent system was normal hexane: methyl alcohol:water（2: 1: 1,V/V）.considering the light phase as stationary phase and heavy phase as mobile phase. Collecting the fraction based on the online chromatogram. And isolating and purifiding its crude extraction applied with Pre-HPLC, its HPLC condition was mobile phase methyl alcohol-0.2%g lacial acetic acid（80:20,V:V）,detector 88232 UV ultraviolet detector;wavelength λ=315 nm,flow velocity=2.8 mL?min^-1, column temperature T=30℃, sample size=20 μL,the molecular mass of target peak is 398.13 by MS, the structure confirmed was caulerpin after being identified by NMR. Studying its content by quantitative Analysis, it came out the content of caulerpin was 0.91 mg·g^-1.