Study On The Extraction,Purification Technology And Activity In Vitro Of Active Components From Aroni Amelanocarpa Fruits

The fruit of the Aroni amelanocarpa was rich in nutrition which contained a large amount of nutrient components such as flavonoids,anthocyanidin,anthocyanin,and so on.It had the functions of resisting inflammation,resisting radiation,reducing blood sugar,protecting liver which could be processed to make health drinks,juice,fruit wine,jam and other foods.The extraction,purification technology,quality study and activity in vitro of both total flavonoids and anthocyanidins from the Aroni amelanocarpa fruit were studied,which provided basis for the further development and application of the Aroni amelanocarpa fruit.Firstly,the extraction technology of total flavonids from the Aroni amelanocarpa fruit was investigated by various extraction methods.The extraction technology was optimized by orthogonal test on the basis of single factor experiments,and the optimal extraction parameters was obtained as follows:ionic liquid dosage of 0.03 g/g?ionic liquid/raw drug?,volume fraction of ethanol of 30%,raw material/liquid ratio of 1:15?g/mL?,microwave power of 450 W,extraction time of 35 min,ultrasonic power of 100 W and ultrasound time of 20 min.Under the optimized conditions,the extraction rate of total flavonoids from the Aroni amelanocarpa fruit was 4.95 mg/g.The extraction technology of anthocyanidins from the Aroni amelanocarpa fruit was studied by ionic liquid assisted ultrasonic technique.Results showed the extraction rate of anthocynidins was improved significantly with the aid of ionic liquid.Brominated-1-butyl-3-methyllimidazole of 0.5 mol/L was selected to be the optimal inonic liquid.The effect of other factors on extraction rate was studied by single factors and orthogonal experiments,80%ethanol aqueous were determined to be the optimum solvent,the ratio of raw material to liquid was 1:25?g/mL?,the extraction time was 3 h,the extraction temperature was 65?and the ultrasonic power was 70 W.Under the above conditions,the extraction rate of anthocyanin can reach 8.82mg/g.Secondly,on the basis of screening of macroporous adsorption resins by static experiments,the macroporous adsorption resins purification technology for active components in the Aroni amelanocarpa fruit was explored using dynamic adsorption and desorption experiments.The optimal purification conditions obtained from single factor experiment was as follows:sample dosage of 0.17 mg/g?raw material/resin?,sample concentration of 40.00?g/mL,pH of 7.0,sample adsorption velocity of 1.5 mL/min,elution solvent of 70%ethanol aqueous and amount of elution solvent of 3 BV.After purification with LSA-21 resin,the content of total flavonoids in the Aroni amelanocarpa fruit crude extract increased from 10.5 mg/g to 56.8 mg/g,indicating a purity enhancement of 5.5 times.And the optimized purification technology of anthocyanins was as follows:the sample dosage was 0.45 mg/g?raw material/resin?,the sample concentration was 55.77?g/mL,the adsorption velocity of 1.5 mL/min,the elution solvent was 50%ethanol aqueous,and the elution volume was 1 BV.After purification with LSA-21 resin,the content of anthocyanins in the Aroni amelanocarpa fruit extract increased from 2.4 mg/g to 19.7 mg/g,meaning a purity inprovment of 8.3 times.Thirdly,the content of total flavonoids,anthocyanins and proanthocyanidins in the Aroni amelanocarpa fruit was determined by UV,in which the methodology was well validated and studied.The content of hyperoside in the Aroni amelanocarpa fruit and cypermethrin chloride in the corresponding extract was determined by high performance liquid chromatography?HPLC?.Results showed the content of hyperoside in the Aroni amelanocarpa fruit was 27.37?g/g,and the content of cypermethrin chloride in the extract was 1.86 mg/mg.In addition,the antioxidation ability of the Aroni amelanocarpoa fruit extract was studied by in vitro antioxidation test.Results showed the Aroni amelanocarpa fruit extract possess certain scavenging activity for DPPH·,·OH,·O₂^-and ABTS·⁺radical,and the corresponding IC₅₀0 were2.65,2.42,0.48,0.66 mg/mL,respectively.Finally,the single factor test and orthogonal test were used to screen the type,proportion,dosage and forming process of the Aroni amelanocarpa fruit extract granule prescription auxiliary materials respectively,and relative quality of the resulting granule were examined.Results indicated the particle size,fluidity and other projects meet the requirements of“Chinese Pharmacopoeia 2015 Edition”.The molding process was simple and feasible,and the stability was ideal.