Study On Separation, Identification And Protective Effect Of Oxidative Damage Induced By Radiation Of Polyphenols In Pinus Koraiensis

Pinus koraiensis are rare plants which mainly produced Xiaoxing'an Mountain in Heilongjiang Province. Pinus koraiensis contains a large amounts of phenolic compounds, including terpenoids, polyphenols, alkaloids and other biologically active ingredients ,which had anti-radiation, anti-oxidation and other biological functions. In this study, the barks of Pinus koraiensis were as the plant materials and studied on its antioxidant and anti-radiation bioactivity. PKB polyphenol were extracted by using ethanol as extraction solvent, and the active ingredients of PKB were extracted in the process and the optimum extraction conditions were optimized. The PKB polyphenol purified by AB-8 macroporous resin for the fisrt time, which was used different gradient elution of ethanol to do the free radical scavenging,iron ion chelating capacity and the total antioxidant capacity. The study was adopted by tracking method with biological activity, and selected the strongest component to do anti-radiation experiments in vitro. And then the PKB polyphenol was purified gradually by using of semi-preparative liquid chromatography for the second time, and finally identified the polyphenol components by HPLC. The main contents of this study were stated as the following:(1) PKB polyphenol extraction solvents were investigated for methanol, ethanol, acetone, ethyl acetate and water on the yield of PKB polyphenol, which is up to 7.78mg / g with ethanol, finally ethanol was choosed as extraction solvents.(2) The ultrasonic extraction method was selected to extract the polyphenol in PKB. The main factors which were impacted on polyphenol yield were ensured by single trial test, and then the response surface method was combined to optimize the polyphenol extract conditions. on the basis of ultrasonic power 160W. The optimum conditions were extraction time 30.61mins, the ratio of material to solvent 1: 30.79, the ethanol concentration 42.20%, the yield of PKB polyphenol was 8.241mg / g.(3) The study was adopted by tracking method with biological activity, The PKB polyphenol purified by AB-8 macroporous resin for the first time, which was using ethanol as extraction eluent to obtain20%, 40%, 60% ethanol elution with different concentrations, and dried into powder. The biological activity in vitro was mainly inspected the hydroxyl radical (?OH),superoxide anions (O^2-?),ABTS radicals and DPPH radicals,iron ion chelating capacity and the total reduction ability to definite the strongest component. Finally the biological activity of 40% ethanol elution was significantly higher than other components. (4) The 40% alcohol elution was purified for the secondary by semi-preparative liquid chromatography to obtain higher purity of PKB polyphenols, which was analyzed its polyphenol components by high performance liquid chromatography. The results showed that the coumaric acid is 321.4±1.0mg/100g, salicylic acid is 117.7±3.1mg/100gand vanillic acid is 101.3±2.8mg/100g, followed by caffeic acid is 54.2±3.2mg/100g and protocatechuic hormone is 25.3±1.9mg/100g, the lowest baicalin is 8.05±1.2mg/100g.(5) When human white blood cells were exposured to ionizing radiation, The impact on the number of white blood cell added the different dose groups of pine polyphenols was significantly higher than the radiation group, there appeared shaped leaf cells, atypical lymphocytes cells and degeneration cells. Pine polyphenols can inhibit the radiation on thymus and liver from damaging, which can significantly increase superoxide dismutase activity and glutathione peroxidase activity, and reduce the peroxidation product MDA.