| Investigation and research were carried out for extraction, qualitative analysis N separation and identification of anthocyanine from fruit of Ericaceae. Vaccinium. Blueberry. The result showed that anthocyandine usual existed in fruit of Blueberry and were high in content. Anthocyanins has been discussed in relation to a wide range of physiological functions such as vision improvement and have inhabition of low density lipoprotein oxidation, hypercholesterolemia, antioxidanU antidecrepitudeN antiulcer, antivirus and anticancer and so on. The author aimed chiefly at developing and utilizing blueberry resource, and exploring it's major compounds. Meanwhile a new method suitable for mass production of anthocyanine was founded.High performace liquid chomatogramphy (HPLC) is a best method of quantitative analysis. Samples were saluted 0.1%HC1 MeOH: Water =1:4 solvent system and determinated by RP-HPLC. Analysic condition of RP-HPLC: Anthocyanins were seperated on a zorbax C18 column(4.6×l50mm); Mobile phase A solvent: MeOH; B solvent: 10% formic acid; flow rate 1.0 mL/min;column temperature : 25℃ ; detector wavelength 520nm , 517nm , 530nm , 525nm;Anthocyanins(20i 1 injected)were eluted with gradient; 0~20min, B Solvent 50~60%,20~25min B solvent 60-100%; 25~30min B solvent 100%.The linear range of CY-3-GLC was 0.018-0.030ug ; DI-CH was 0.160-0.800ug; OE-CH was 0.044~0.440|ig ; MA-GA was 0.224-0.112|ig . The average recoverage rate was 98.58%, 96.34%> 95.45% and 97.93% respectively, and all were above 90.00%, so that the result was accurate. Four standard exponent of anthocyanine content and total content of its were measured in the same condition.The quantitative method for blueberry anthocyanine was established with color value and high performace liquid chomatogramphy. First,antohcyanine content differed among samples(cyandine-3-glucoside -> cyandine-3-galactoside -. malvidine-3-glucoside> malvidine-3-galactoside) with color value; Second, four kinds of anthocyanine content and total its content were precisely measured in five varieties with RP-HPLC. Conclusion was drawn that the order of total anthocyanine was Chignecttx CA-206, NB-3> Bog bilberry and Lingonberry. and 725.413 mg/lOOg fresh fruit> 357.224 mg/lOOg fresh fruit> 350.380mg/100g fresh fruit> 343.669mg/100g fresh fruit, 147.759 mg/100g fresh fruit respectively. Moreover, the category and content were different in fruit of five samples. Cyandine-3-galactoside was an usual anthocyanine, but cyandine-3-glucoside didn't exsit in Bogbilberry and Lingonberry, the fruit of Lingonberry contained one of four kinds of anthocyanine, which was cyandine-3-galactoside. As a result, genotypes were diverse in species and varieties, which result in different category and content of anthocyanine in fruit of blueberry.In order to study the course of extract and separation, Lingonberry was a sample,and squeezed. The juice was saluted with 85% ethanol for 3 hours in 60℃, and condensed. NKA-12 microreticular resin was used firstly in the technique to extract and separate anthocyanine with water and 30%, 50%, 90% ethanol as solution. Then, polyamide chromatogramphic column was used for seperation in the same method. And compound I was extracted.Compound I was identified by physical and chemical assay. Fused point: 181-182℃; Soluble water, easy soluble MeOH, difficult soluble acetone and dark-red power. Thin layer chromatogramplK HPLC, Mass spectroscopy and 13C nuclear magnetic resource spectroscopy. It was identified as cyanidene - 3 -galactoside (2-3' , 4' - dihydroxy benzene-5, 7-dihydroxy pyran-3-O - P -D-galactoside); Molecular weight 449; and molecular formula: C2iH2iOn.The paper provides a scientific basis for researching and developing blueberry anthocyanine as effective ingredients, exploiting and utilizing blueberry resource for further, especially mass production of extract and separation, and creating economic and social benefits. |
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