Study On Anthocyanins Composition And Bioactivities Of Berries

Phenolic compounds is the most important secondary metabolism substance and widely distributed in the plant.Berries are rich in phenolics,especially anthocyanins.It is widely in cells of flowers,fruits,stems,leaves and roots,and responsible for color of fruits,plants and flowers,such as red,orange,purple and blue.For its unique biological activity,it was applied to remove free radicals,anti-inflammatory,prevent diabetes,hepatoprotective effect,inhibition of lipid peroxidation and platelet aggregation,antitumor,protect vision,prevention of alzheimer’s disease and so on.Because anthocyanins are mostly water-soluble pigments,and its structure are complexity and instability,anthocyanins have some difficulties in extraction,isolation and purification.In this research,the identification and biological activity of anthocyanin in four kinds of berries(blueberry,bilberry,mulberry,and cranberry)were studied,and the correlation and difference between the chemical composition and biological activity were also revealed.The composition of black raspberry,which were rich in cyanidin,were systematically purification,identified,and the bioactivities were also evaluated,which can make a new application value for the anthocyanins of berries.This is the first time to identify the anthocyanins from four kinds of berries by Fourier transform-ion cyclotron resonance mass spectrometry(FT-ICR MS)and evalute their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B.The polyphenolic profiles of four berries in China were investigated by FT-ICR MS and 39 polyphenols including 26 anthocyanins,9 flavonoids,and 4 phenolic acids were identified accuratly.The most anthocyanin,about 24 were identified in blueberry,and bilberry(17),cranberry(13),and mulberry(8).Cyanidin are common in four berries,and cyanidin-3-O-glucoside was the highest in 4 berries.Malvidin are the main compounds in bilberry and cranberry.Catechin,and derivatives of myricetin and quercetin are the most in flavonoids.Protocatechuic acid and chlorogenic acid are most in phenolic acids.The anthocyanin level in blueberry are the highest and showed the strongest antioxidant and PTP1B inhibition capacities.In α-glycosidase,the mulberry extraction presented strongest inhibition capacity,which was rich in flavonoids.Molecular docking results showed that cyanidin aglycones had the highest inhibition activity to PTP1B among 6 type of anthocyanins.Polyphenolic profile as well as anti-oxidant and anti-diabetes effects of extracts from freeze-dried black raspberries was tested by LC-MS and α-glycosidase,PTP1B inhibition assays.And the results firstly showed cyanidin-glucoside had good PTP1B inhibition.Twenty polyphenolic profiles(5 anthocyanins and 15 phenolics)were identified by HPLC-MS/MS.10 compounds(3 anthocyanins and 7 phenolics)were isolated and confirmed by NMR,and two were isolated from Rubus for the first time.Compound 8(Cyanidin-3-O-glucoside),9{Cyanidin-3-[(2"-O-xylosyl)-(6"-O-rhamosyl)]-O-glucoside},11(Cyanidin-3-O-rutinoside)are anthocyanins,compound 2(4-O-β-D-glucopyranosyl benzoate),3(Protocatechuic acid),12(Epicatechin),13((E)-8-β-Dglucopyranosylcinnamic acid),16(Quercetin-rutinoside),17(Ellagic acid)and 20(Protocatechuic acid methylester)are phenolics.Freeze-dried black raspberry powder extracts were purified by polyamide resin with different concentrations,and 30%fractions,compound 8,9,11,17 exhibited optimal antioxidant activity.In a-glycosidase assay,the 70%fraction and compound 16,17 demonstrated the greatest inhibitory effect.In PTP1B assay,the 30%fraction and compound 8,9,11,16,17 exhibited the highest inhibitory effect.The anthocyanins from black raspberry showed hepatoprotective effect on subacute and acute alcohol-induced hepatic injury.Co-administration of BRAs with alcohol significantly suppressed the rising of contents of AST,ALT,CHOL,TBIL,LDL in serum and liver index.These results were accompanied by histological observations of liver tissue.MTT assay was performed to evaluate cell cytotoxic activity on t-HSC/Cl-16,HepG2 and Hep3B,and the results demonstrate that cyanidin has anticancer effect and show no cell cytotoxic on nomal cells.