Study On The Utilization Of Polyprenols From Solid Waste In The Production Process Of Ginkgolides Injection

This dissertation was supported by Jiangsu environmental protection research project(2015022)—Part experiment content of "Demonstration research of recycling utilization on solid waste from production process of Reduning and ginkgolide injection"The dissertation was divided into six chapters.Chapter one,literature researchThe utilization status of Ginkgo biloba were reviewed including resources chemistry and the utilization of Ginkgo biloba polyprenols.The leaves of Ginkgo biloba are rich in flavonoids,ginkgo terpene lactones,ginkgo phenolic acids,steroids and polyprenols,which possess biological activities of anti-oxidation,anti-tumor,dilating blood vessels,anti-inflammatory,improving microcirculation.In this chapter,resource distribution,chemical composition,pharmacological action,clinical application,product development of Ginkgo biloba and utilization of the resources of polyprenols were analyzed.As a result,this review would provide reference for comprehensive utilization of Ginkgo leaf,and the resource utilization of ginkgo biloba residue while production process.Chapter two,study on the extraction and purification of the polyprenols and the total flavonoids from solid waste in the production process of ginkgolides injectionThe extraction of the polyprenols from solid waste in the production process of Ginkgolides injection was optimized by orthogonal method.The parameters included soaking time,extraction time,extraction temperature and particle size.As well as the hydrolysis process was also optimized by ultrasonic time,ratio of alcohol to water,concentration of KOH and ultrasonic frequency.The high purity polyprenols was obtained by acetone cold separation,silica gel column impurity removal and C18 column impurity removal.Subsequently,the high purity of polyprenols was obtained.The extraction liquid was added with an equal amount of 2%KOH methanol water(1:4)solution,and hydrolyzed for 20 min under 100KHz condition.The purity of the purified polyprenols was 95%.The content of total flavonoids in Ginkgo biloba residue was determined by spectrophotometry.The extraction process of total flavonoids in Ginkgo biloba residue was optimized by orthogonal method,and obtained the best extraction process was taken,with 20 times the amount of absolute ethanol added,then heat reflux extraction 0.5 h.Chapter three,quality standard research of polyprenols from ginkgo leafThe polyprenols possess important physiological and pharmacological activities,and some new drugs have been listed abroad,For the better comprehensive utilization of polyprenols in Ginkgo biloba,the quality standard of polyprenols in ginkgo residue was carried out,including the qualitative analysis of polyprenols by TLC and quantitative analysis by HPLC.The quality control includes the appearance and odor,solubility,contents determination,moisture determination,solution color and heavy metal inspection,which could qualitative and quantitative analysis the polyprenols in Ginkgo biloba and identified its structural characteristics.Chapter four,pharmacological activity evaluation of polyprenols in Ginkgo leafIn order to investigate the effect of Ginkgo biloba polyphenol on acute liver injury,the acute liver injury in rats was induced by CCl4,and the biphenyl diester was used as positive drug.The changes of ALT,AST,ALP,ALB,liver and colon in serum and the changes of beneficial bacteria(lactobacillus,bifidobacteria),harmful bacteria(enterococci,enterobacteria)in intestinal contents were investigated.In acute liver injury rats,ALT?AST?ALP level were increased obviously,and the content of ALB decreased significantly,pathological changes appeared in liver and colon,contents of beneficial bacteria decreased,and harmful bacteria increased.After polyprenols administration,contents of ALT,AST,ALP decreased while ALB increased,the pathological changes of liver and colon reduced and intestinal contents of bacteria increased,harmful bacteria decreased.Chapter five,study on the mechanism of liver protection of polyprenols in ginkgo leafUPLC-MS was applied for metabolomics analysis of serum in rats with acute liver injury induced by CC14,and the changes of endogenous metabolites in serum of rats with acute liver injury were studied.By comparing the changes of endogenous metabolites in serum of normal group,model group and administration group,the potential drug action markers were found,and the intervention mechanism was analyzed preliminarily.The results showed that the liver lesion causes the abnormal of tryptophan acts,than lead to all kinds of pathological symptoms.After the intervention of polyphenol,the tryptophan acts return to normalThe 16s rRNA technology was applied in the study of the diversity of intestinal flora intestinal contents of rat intestinal flora diversity in acute liver injury rats and normal rats,which indicated polyprenols can obviously improve the internal environment and promote the restoration of intestinal microflora.Chapter six,biotransformation of polyprenolsAccording to the literature reports,the physiological activity of isoamyl alcohol derivatives is stronger than that of polyprenols.Nowadays,the transformation of polyprenols were mainly for chemical conversion,in consideration of the ginkgo leaf decomposed by microorganism in soil,in this section,the rhizosphere soil and surface soil of Ginkgo biloba in different locations and different media were sampled and screened.Different cultures were used to screen different types of bacteria.There are 14 kinds of fungi and 11 fungi,of which 13 are common bacteria,9 are fungi,1 are against organic solvents,and 2 are fungi.The transformation of Ginkgo biloba polyprenol was carried out by using the sieved strain.The results may be affected by the amount of organic solvent,and the range of the sample was too small to obtain the microbial for polyprenols transformation.