抗氧化活性测定方法.docVIP

抗氧化方法 Determination of Superoxide Anion Scavenging Ability（超氧阴离子清除能力） determined by a CL method in the pyrogallol-luminol system on a BPCL Ultra-weak luminescence analyzer。（焦性没食子酸-发光胺，荧光检测） 步骤：10?L sample (不同浓度) + 50??L焦棓酸(6.25*10-4 mol/L) + 0.94 mL of a mixture containing luminol (0.05 mol/L) and carbonate buffer (pH 10.2) （发光胺用pH 10.2碳酸盐缓冲液配成0.05 mol/L） Hi-V, 800; Kv, -1; the spectral range of CL（波长范围）180-800 nm; 温度：30 ° C.总时间：300S，每2S读数一次。 对照：不加样品（样品用水代替）。空白不加焦棓酸（用来调零）。 二、Determination of Scavenging Ability on Hydroxide Radicals（羟基自由基清除能力） 1、CuSO4-Phen-Vc-H2O2 CL system （1 mL体系） 2、50??L of sample solution （样品液） + 50 ?L of a 1.0 mmol/L CuSO4 solution （CuSO4 溶液）+ 50 ?L of a 1 mmol/L 1,10-phenanthroline solution（邻二氮菲溶液）+ 700 ??L of a 0.05mol/L borate buffer (pH 9.0) （硼酸缓冲液）+ 100 ?L of a 1 mmol/L ascorbate solution + 50 ?L of a 0.15% H2O2 solution 3、总时间400S，间隔3S。Hi-V, 800; Kv, -1; the spectral range of CL（波长范围）180-800 nm; 温度：30 ° C. 4、阳性对照：不加样品（样品用水代替）。空白不加H2O2（用来调零）。 Determination of Scavenging Effect on Hydrogen Peroxide（过氧化氢清除能力） 1、luminol-H2O2 system 2、The luminescent reaction was initiated by manually adding 1 mL of a solution containing 0.15 mol/L hydrogen peroxide and 0.1mol/L luminol per liter of carbonate buffer (0.05 mol/L, pH 9.4). Light emission vs time was recorded for 3 min at 2 s intervals. 步骤：0.15 mol/L的过氧化氢 + 0.1mol/L发光胺（用pH 9.4的碳酸缓冲液配），总体系1ml. 3、BPCL Ultra-weak luminescence analyzer，Hi-V, 800; Kv, -1; the spectral range of CL（波长范围）180-800 nm; 温度：30 ° C. 4、The control was performed in the same manner in the mixture without the sample solution, and the background was detected without H2O2 addition。 Determination of Preventing DNA Damage Effect（DNA损伤） 1、CuSO4-Phen-Vc-H2O2-DNA CL system. 步骤：Copper and 1,10-phenanthroline were premixed in 0.1 mol/L NaOAc/HOAc (pH 5.5) buffer。 3 ?g/mL DNA was incubated with a phen-Cu solution. 步骤：800 ?L of phen-Cu/DNA solution + 100 ?L of 4.2 *10-3 mol/L ascorbate + 200 ?L of 6% H2O2 + were added without interval t