Investigation On Separation, Purification And Biological Activity Of Anthocyanins And Polyphenols In Blueberry

Blueberry anthocyanins and polyphenols were chosen as research objects.Different extraction processes were taken to separate the two materials. AB-8macroporous resin was used to pure blueberry anthocyanins and polyphenols crudeextracts. At last, four kinds of blueberry extracts, blueberry anthocyanins crude extract,polyphenols crude extract, purified anthocyanins and purified polyphenols wereobtained. Studies of the four extracts on vitro antioxidant, anti-tumor in vitro,anti-tumor in vivo and immune function were carried out.Three methods, ferric reducing antioxidant power (FRAP), the DPPH methodand abts radical scavenging activity (ABTS), were taken to compare antioxidantactivity of the four extracts. The results showed that, the antioxidant activity ofpurified anthocyanins and polyphenols are significantly increased (P<0.05).Anthocyanins extracts have higher antioxidant activity than polyphenols.MTT assay was used to analysis the inhibiting abilities of the four extracts onbreast cancer cells (MDA-MB231) in vitro. Results showed that the improvement ofthe antioxidant capacity of the two purified product did not improve their abilities toinhibit tumor proliferation in vitro, on the contrary, anthocyanins and polyphenolscrude extract had higher ability of inhibiting tumor proliferation (P <0.05).Based on vitro experiments results and with cyclophosphamide as positivecontrol, the four extracts were applied to do anti-tumor experiment of CD-1breastcancer model mouse. The results showed that the four extracts of the three dosegroups of tumor suppression capability although lower than positive control, but allhave certain degree of inhibition. They can also significantly improve indicators oftumor suppression, body weight, thymus&spleen index, macrophages phagocytosisand lymphocyte transformation capacity and SOD activity (P <0.05), and greatlyreduce the accumulation of MDA in lung and liver and NO in serum (P <0.05).Polyphenols and anthocyanins crude extract mixture works best. So the blueberryextracts can inhibit tumor growth effectively, while protect mice bodily functions.Material composition and structure analysis of purified blueberry anthocyaninsand phenolic compounds were carried out though high performance liquidchromatography-mass spectrometry (HPLC-MS). Found that, in anthocyanins mainlycontain pelargonidin-3-O-anthocyanin galactoside, quercetin-3-O-galactoside and mallow pigment-3-O-glucose, etc. In polyphenols mainly containquercetin-3-O-galactoside and epicatechin and other substances. In these compounds,majority contains of the3-O-hexose or pentose glycoside structure.The findings lay the theoretical foundation for a comprehensive evaluation ofblueberry anthocyanins and polyphenols activity and reveal their anti-tumormechanism.