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目的研究生物活性物质体外抗氧化能力评价方法,应用体外理化环境下建立起来的评价方法对受试物进行初步抗氧化能力评估。方法联合应用2,2’-连氮-双-(3-乙基苯并噻唑-6-磺酸)二铵盐(ABTS)法与铁离子还原/抗氧化能力测定法(FRAP)法用于生物活性物质体外抗氧化能力研究,在ABTS法中,使用紫外可见分光光度计734nm波长下测定以槲皮素、姜黄素、DL-α-生育酚和原花青素等为代表的生物活性物质;在FRAP法中,运用酶标仪,595nm波长处测定同样受试物的抗氧化能力。结果ABTS法:槲皮素和姜黄素的抗氧化活性TEAC值分别约为2.02和0.50;而1gDL-α-生育酚、原花青素清除自由基的能力相当于2.06mmol、2.897mmol的Trolox清除自由基的能力;FRAP法:抗氧化活性以1.0mmol/L FeSO4为参考标准,槲皮素、姜黄素和Trolox摩尔当量约为5.73、1.18和2.09;而DL-α-生育酚和原花青素抗氧化活性分别是207.7mg、156.36mg。结论ABTS法与FRAP法联合应用,操作简便、结果可靠,尤其适宜作为生物活性物质体外抗氧化能力的评价方法。
OBJECTIVE To study the in vitro anti-oxidative capacity of bioactive substances, and to evaluate the preliminary anti-oxidative ability of the tested substances using the evaluation method established in vitro and in physical and chemical environment. Methods A combination of 2,2’-azinobis- (3-ethylbenzothiazole-6-sulfonic acid) diammonium salt (ABTS) and ferric ion reduction / antioxidant capacity assay Bioactive substances in vitro antioxidant capacity in the ABTS method, the use of UV-visible spectrophotometer at 734nm wavelength determination of quercetin, curcumin, DL-α-tocopherol and proanthocyanidins and other bioactive substances represented by the FRAP France, the use of microplate reader, 595nm wavelength determination of the same test substance antioxidant capacity. Results The ABTS method showed that the TEAC values of quercetin and curcumin were about 2.02 and 0.50, respectively. While 1gDL-α-tocopherol and proanthocyanidin scavenged free radicals, their abilities to scavenge free radicals were 2.06mmol and 2.897mmol respectively. Ability; FRAP method: antioxidant activity 1.0mmol / L FeSO4 as a reference standard, quercetin, curcumin and Trolox molar equivalents of about 5.73,1.18 and 2.09; and DL-α-tocopherol and proanthocyanidins antioxidant activity were 207.7 mg, 156.36 mg. Conclusion The combination of ABTS and FRAP has the advantages of simple operation and reliable results, and is particularly suitable as an evaluation method for the in vitro antioxidant capacity of bioactive substances.