Separatation And Analysis Of Several Important Biological Activities In Ginkgo Biloba

The latest research shows that biflavone has antioxidant,antiviral and anticancer activity,but there is still a lack of standard sample and biflavone detection method.In addition,one component of flavonoids and terpenoids have also become a research hotspot of international drug development.Consequently,this thesis with Ginkgo biloba extract as starting material,biflavone,ginkgo acid,flavonoid,etc.were separated and purified.Then single component biflavone standard sample,one component of flavonoid aglycone,terpenoid and ginkgolic acid were prepared by C18 column with NMR analysis.Finally,the 11 kinds of mg grade high purity single component material were isolated,the chromatography conditions were explored and the preparation process was optimized.(1)On the premise of optimizing the separation technology of ginkgo biloba extract,the preparation sample of biflavonoid with total content of 80.4% was obtained by antisolvent crystallization and the recovery rate was 62.9%.Four kinds of monocomponent bioflavonoids: amentoflavone 31 mg,bilobetin 38 mg,sciadopitysin 51 mg and the mixture of ginkgetin and isoginkgetin 87 mg were obtained by chromatographic separation from the prepared samples,and the total yield was 87.1%.The purity was obtained by the method of peak area normalization and their purity were 98.45%,98.66%,98.87% and 99.29%,respectively.The total content of biflavone in ginkgo,ginkgo biloba leaves and exocarp were 0.01 mg/g,6.98 mg/g and 2.31 mg/g,respectively.The optimum chromatographic conditions for the preparation of single component bisflavonoids were as follows: adding 0.005 mol/L glacial acetic acid and trifluoroacetic acid to improve the peak shape,respectively,gradient elution was applied from 76% methanol solution to methanol,elution rate 20 mL/min,sample amount 10 mL,separating 2 times.(2)Three kinds of one component flavonoid glycosides were obtained by chromatography.Quercetin 28.59 mg,kaempferol 22.87 mg and isorhamnetin 4.92 mg,the purity of those were 99.8%,99.2% and 99.7%,respectively,the total yield was 89.4%.The optimum chromatographic conditions were as follows: 0.005 mol/L glacial acetic acid and 0.005 mol/L trifluoroacetic acid l were added to the mobile phase of 60% methanol,the elution rate was 20 mL/min,the sample amount was 20 mL,and separating 2 times.(3)The purified ginkgolic acid was separated,and four single components of ginkgolic acid were prepared: hydrogenated ginkgolic acid 203.3 mg,ginkgo phenol 30 mg,albino acid 121.3 mg,ginkgo diphenol 10.1 mg,total 337.7 mg.The yield of Ginkgo biloba was 84.5%,and the chromatographic purity was 96.2% 94.2% 94.1% 98.3%.The optimum chromatographic conditions for the preparation of single component ginkgolic acid were obtained.For the isometric elution of 1: 92% methanol solution,the elution rate was 18 mL / min,the sample amount was 10 mL,and the separation time was 1.(4)The crude product of terpene lactone was purified by macroporous resin,and the sample of terpene lactone with purity of 85.1% was obtained in the yield of 75.2%.The terpene lactone was prepared by C-18 column.A mixture of bilobalide and ginkgolide C was obtained with a purity of 90.4%,a mass of 61.1 mg and a yield of 42.6%.The mixture of ginkgolide B and ginkgolide A was obtained with a purity of 85%,a mass of 50.7 mg and a yield of 35.3%.The chromatographic conditions were as follows: gradient elution was applied from 20% methanol to 60% methanol gradient elution,elution speed 20 mL/min,sample amount 10 mL,separation time was 1.