几种黄酮类化合物清除DPPH自由基微量模型的建立
摘要
本论文主要研究了三种黄酮类化合物,即黄酮醇、黄酮苷和黄烷醇,清除DPPH自由基微量模型的建立,大致可分为五个部分。
第一部分为前言,概述了抗氧化和黄酮类化合物的研究现状,并引进了更为客观的抗氧化活性评价指标TEAC,即Trolox当量。
第二部分是黄酮醇类化合物清除DPPH自由基微量模型的建立,以槲皮素为研究对象,通过单因素考察,建立L_9(3~4)正交试验,对正交试验结果进行直观分析和方差分析,确定最适实验条件是A_3B_1D_2,即DPPH的最佳浓度为200μmol/L,加入量为175μL,反应20 min。验证实验RSD为3.17%,说明该方法稳定可靠;标准品Trolox、PG、BHA和BHT对照,证明该模型准确可行。实验结果证明,用槲皮素建立的DPPH抗氧化微量筛选模型快速,准确,用样量小,且稳定可靠,适合于槲皮素类似的天然产物抗氧化活性的快速筛选。
第三部分是黄酮苷类化合物清除DPPH自由基微量模型的建立,以芦丁为研究对象,通过单因素考察,建立L_9(3~4)正交试验,对正交试验结果进行直观分析和方差分析,确定最适试验条件是A_3B_1D_2,即DPPH的浓度为250μmol/L,加入量为150μL,反应40 min。验证实验RSD为2.87%,表明该方法稳定可靠;标准品Trolox、PG,BHA和BHT对照实验,表明该模型准确可行。实验结果证明,用芦丁建立的DPPH抗氧化微量筛选模型快速,准确,用样量小,且稳定可靠,适合于芦丁类似的天然产物抗氧化活性的快速筛选。
第四部分是黄烷醇类化合物清除DPPH自由基微量模型的建立,以儿茶素为研究对象,通过单因素考察,建立L_9(3~4)正交试验,对结果进行直观分析和方差分析,确定最适实验条件是A_3B_1D_2,即DPPH的最佳浓度为250μmol/L,加入量为125μL,反应20 min。验证实验RSD为1.06%,表明该方法稳定可靠;标准品Trolox、PG、BHA和BHT作对照,表明该模型的准确性。实验结果证明,用儿茶素建立的DPPH抗氧化微量筛选模型快速,准确,用样量小,且稳定可靠,适合儿茶素类似的天然产物抗氧化活性筛选使用。
第五部分是文献综述,综述了国内外有关抗氧化研究与防治疾病的关系,抗氧化的研究方法,DPPH方法的作用机理以及DPPH方法的研究和应用现状。
This paper mainly studies three microplate-based screening methods of DPPH antioxidant assay for three kinds of flavonoids, which are flavonols, flavonol glycosides and flavanols. And the paper is composed of five components.
The first part is the preface, which summarizes the research of antioxidant and flavonoids, as well as antioxidant activity evaluation TEAC, that is to say Trolox equivalent antioxidant capacity.
The second part is the microplate-based screening methods for quercetin of DPPH antioxidant assay. To establish an antioxidant model of microplate assay in vitro by quercetin, the orthogonal experiment was designed according to L_9(3~4) orthogonal table after determining the single factor exploration. The optimum experiment condition, which was A_3B_1D_2, that was, the concentration of DPPH was 200μmol/L, the addition volume of DPPH was 175μL and kept 20 min in room temperature, was determined by the direct-viewing analysis and variance analysis of the orthogonal experimental results. RSD of the evaluation test was 3.17%, showed that the method was stable. And the standard of Trolox, PG, BHA and BHT as control further certified the precision and feasibility of the model. The results indicated that the microplate assay was rapid, accurate, little sample, stable and reliable for the screening antioxidant activity of the quercetin similar traditional Chinese medicine.
The third part is the microplate-based screening methods for rutin of DPPH antioxidant assay. To establish an antioxidant model of microplate assay in vitro by rutin, the orthogonal experiment was designed according to L_9(3~4) orthogonal table after determining the single factor exploration. The optimum experiment condition , which was A_3B_1D_2, that was, the concentration of DPPH was 250μmol/L, the addition volume of DPPH was 150μL and kept 40 min in room temperature, was determined by the direct-viewing analysis and variance analysis of the orthogonal experimental results. RSD of the evaluation test was 2.87%, showed the method was stable. And the standard of Trolox, PG, BHA and BHT as control further certified the precision and feasibility of the model. The results indicated that the microplate assay of rutin was rapid, accurate, little sample, stable and reliable for the screening antioxidant activity of the rutin similar natural products.
The fourth part is the microplate-based screening methods for catechin of DPPH antioxidant assay. To establish an antioxidant model of microplate assay in vitro by catechin, the orthogonal experiment was designed according to L_9(3~4) orthogonal table after determining the single factor exploration. The optimum experiment condition, which was A_3B_1D_2, that was, the concentration of DPPH was 250μmol/L, the addition volume of DPPH was 125μL and kept 20 min in room temperature, was determined by the direct-viewing analysis and variance analysis of the orthogonal experimental results. RSD of the evaluation experiment was 1.06%, showed that the method was stable . And the standard of Trolox, PG, BHA and BHT as control further certified the precision and feasibility of the model. The results indicated that the microplate assay was rapid, accurate, little sample, stable and reliable for the screening antioxidant activity of the catechin similar traditional Chinese medicine.
The last part is the literature review, which includes the relevant something of the relationship between antioxidant research and disease prevention at home and abroad , the research methods of antioxidant, the mechanism and the condition of studying and application of the DPPH method .
第一部分为前言,概述了抗氧化和黄酮类化合物的研究现状,并引进了更为客观的抗氧化活性评价指标TEAC,即Trolox当量。
第二部分是黄酮醇类化合物清除DPPH自由基微量模型的建立,以槲皮素为研究对象,通过单因素考察,建立L_9(3~4)正交试验,对正交试验结果进行直观分析和方差分析,确定最适实验条件是A_3B_1D_2,即DPPH的最佳浓度为200μmol/L,加入量为175μL,反应20 min。验证实验RSD为3.17%,说明该方法稳定可靠;标准品Trolox、PG、BHA和BHT对照,证明该模型准确可行。实验结果证明,用槲皮素建立的DPPH抗氧化微量筛选模型快速,准确,用样量小,且稳定可靠,适合于槲皮素类似的天然产物抗氧化活性的快速筛选。
第三部分是黄酮苷类化合物清除DPPH自由基微量模型的建立,以芦丁为研究对象,通过单因素考察,建立L_9(3~4)正交试验,对正交试验结果进行直观分析和方差分析,确定最适试验条件是A_3B_1D_2,即DPPH的浓度为250μmol/L,加入量为150μL,反应40 min。验证实验RSD为2.87%,表明该方法稳定可靠;标准品Trolox、PG,BHA和BHT对照实验,表明该模型准确可行。实验结果证明,用芦丁建立的DPPH抗氧化微量筛选模型快速,准确,用样量小,且稳定可靠,适合于芦丁类似的天然产物抗氧化活性的快速筛选。
第四部分是黄烷醇类化合物清除DPPH自由基微量模型的建立,以儿茶素为研究对象,通过单因素考察,建立L_9(3~4)正交试验,对结果进行直观分析和方差分析,确定最适实验条件是A_3B_1D_2,即DPPH的最佳浓度为250μmol/L,加入量为125μL,反应20 min。验证实验RSD为1.06%,表明该方法稳定可靠;标准品Trolox、PG、BHA和BHT作对照,表明该模型的准确性。实验结果证明,用儿茶素建立的DPPH抗氧化微量筛选模型快速,准确,用样量小,且稳定可靠,适合儿茶素类似的天然产物抗氧化活性筛选使用。
第五部分是文献综述,综述了国内外有关抗氧化研究与防治疾病的关系,抗氧化的研究方法,DPPH方法的作用机理以及DPPH方法的研究和应用现状。
This paper mainly studies three microplate-based screening methods of DPPH antioxidant assay for three kinds of flavonoids, which are flavonols, flavonol glycosides and flavanols. And the paper is composed of five components.
The first part is the preface, which summarizes the research of antioxidant and flavonoids, as well as antioxidant activity evaluation TEAC, that is to say Trolox equivalent antioxidant capacity.
The second part is the microplate-based screening methods for quercetin of DPPH antioxidant assay. To establish an antioxidant model of microplate assay in vitro by quercetin, the orthogonal experiment was designed according to L_9(3~4) orthogonal table after determining the single factor exploration. The optimum experiment condition, which was A_3B_1D_2, that was, the concentration of DPPH was 200μmol/L, the addition volume of DPPH was 175μL and kept 20 min in room temperature, was determined by the direct-viewing analysis and variance analysis of the orthogonal experimental results. RSD of the evaluation test was 3.17%, showed that the method was stable. And the standard of Trolox, PG, BHA and BHT as control further certified the precision and feasibility of the model. The results indicated that the microplate assay was rapid, accurate, little sample, stable and reliable for the screening antioxidant activity of the quercetin similar traditional Chinese medicine.
The third part is the microplate-based screening methods for rutin of DPPH antioxidant assay. To establish an antioxidant model of microplate assay in vitro by rutin, the orthogonal experiment was designed according to L_9(3~4) orthogonal table after determining the single factor exploration. The optimum experiment condition , which was A_3B_1D_2, that was, the concentration of DPPH was 250μmol/L, the addition volume of DPPH was 150μL and kept 40 min in room temperature, was determined by the direct-viewing analysis and variance analysis of the orthogonal experimental results. RSD of the evaluation test was 2.87%, showed the method was stable. And the standard of Trolox, PG, BHA and BHT as control further certified the precision and feasibility of the model. The results indicated that the microplate assay of rutin was rapid, accurate, little sample, stable and reliable for the screening antioxidant activity of the rutin similar natural products.
The fourth part is the microplate-based screening methods for catechin of DPPH antioxidant assay. To establish an antioxidant model of microplate assay in vitro by catechin, the orthogonal experiment was designed according to L_9(3~4) orthogonal table after determining the single factor exploration. The optimum experiment condition, which was A_3B_1D_2, that was, the concentration of DPPH was 250μmol/L, the addition volume of DPPH was 125μL and kept 20 min in room temperature, was determined by the direct-viewing analysis and variance analysis of the orthogonal experimental results. RSD of the evaluation experiment was 1.06%, showed that the method was stable . And the standard of Trolox, PG, BHA and BHT as control further certified the precision and feasibility of the model. The results indicated that the microplate assay was rapid, accurate, little sample, stable and reliable for the screening antioxidant activity of the catechin similar traditional Chinese medicine.
The last part is the literature review, which includes the relevant something of the relationship between antioxidant research and disease prevention at home and abroad , the research methods of antioxidant, the mechanism and the condition of studying and application of the DPPH method .
引文
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