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荧光猝灭法测定高粱原花青素-金属离子络合反应     被引量:4

The determining of complexation reaction between Sorghum bicolor Moench proanthocyanidin and metal ions by method of fluorescence quenching

文献类型:期刊文献

中文题名:荧光猝灭法测定高粱原花青素-金属离子络合反应

英文题名:The determining of complexation reaction between Sorghum bicolor Moench proanthocyanidin and metal ions by method of fluorescence quenching

作者:张亮亮[1] 徐曼[1] 汪咏梅[1] 胡新宇[1]

第一作者:张亮亮

机构:[1]中国林业科学研究院林产化学工业研究所

年份:2016

卷号:1

期号:6

起止页码:58-63

中文期刊名:林业工程学报

外文期刊名:Journal of Forestry Engineering

收录:CSTPCD;;北大核心:【北大核心2014】;

基金:国家重点研发计划(2016YFD0600806)

语种:中文

中文关键词:荧光猝灭法;原花青素;金属离子;化学计量比;络合常数

外文关键词:Fluorescence quenching; proanthocyanidin ; metal ions ; stoichiometry ; Stern-Volmer constants

分类号:Q946.84

摘要:原花青素是一种广泛存在于高等植物中具有生物活性的多酚类化合物,络合金属离子是原花青素的基本特性之一,不管是低聚体原花青素还是高聚体原花青素均能够络合金属离子。笔者研究利用金属离子与原花青素酚羟基的络合反应导致的原花青素(PC)荧光猝灭作用,研究高粱原花青素与金属离子的络合反应。移取2 900μL醋酸盐缓冲溶液(pH 6)于1 cm石英池中,加入100μL浓度为05 mmol/L高粱原花青素溶液,再逐次加入浓度为05 mmol/L金属离子(Al^(3+)、Cu^(2+)、Sn^(2+)和Zn^(2+))溶液进行荧光滴定反应,每次加入金属离子溶液量为1μL,金属离子加入总量为10μL。记录原花青素的荧光强度衰变曲线,利用Stern-Volmer荧光猝灭方程计算得到4种金属离子Al3+、Cu^(2+)、Sn^(2+)、Zn^(2+)分别与PC络合常数为135,032,144和029(×10~6L/mol)。利用Jobs法测定得到4种金属离子Al^(3+)、Cu^(2+)、Sn^(2+)和Zn^(2+)与PC形成的金属离子-PC络合物化学计量比(mol∶mol)分别为1∶2,1∶2,1∶1和1∶2。此次对金属离子-PC络合反应定量关系的研究将有助于更好地了解原花青素的生理学功能。
Proanthocyanidins (epicatechin16(4→8) catechin, PC) from Sorghum bicolor Moench are a class of polyphenol compounds widely distributed in higher plants with good physiological and pharmacological effects. It is well known that both procyanidin monomers and oligomers have the capacity to chelate with metal ions. However the studies on the metal-chelating capacity of proanthocyanidins with high molecular weight have been little reported hitherto due to the dif- ficulty in obtaining these proanthocyanidins from the natural products. The fluorescence quenching effects of polyphenolic compounds epicatechin16(4→8) eatechin by metal cations Al^3+ , Cu^2+, Sn^2+ and Zn^2+ were studied at pH 6 by measuring fluorescence quenching spectra. To measure fluorescence, 1 μL of 0. 5 mmol/L metal ions solution were titrated into 100 μL of 0.5 mmol/L polyphenolic compounds solution in 2 900 μL pH 6 acetate buffer solution. The volume of metal ions solution was varied from 0 μL to 10 μL with the final concentration of 1.67 μmol/L. The results indicated that an obvious quenching effect on PC fluorescence intensity by Al^3+, Cu^2+, Sn^2+and Zn^2+ was observed, indicating the interaction between metal ions and PC. Continue adding more Sn〉 , Al^3+, Cu^2+ and Zn^2+ ions into solution caused a gradual de- crease in the PC fluorescence intensity without changes of the maximum emission wavelength and shape of the peaks, which was interpreted that Al^3+, Cu^2+, Sn^2+ and Zn^2+ could quench the intrinsic fluorescence of PC. The Stern-Volmer equation was used to assessment of the Stern-Volmer constants (Ksv) between metal ions and PC. The results shown the metal Stern-Volmer constants of metal ions Al^3+, Cu^2+, Sn^2+, Zn^2+ for PC were 1.35, 0. 32, 1.44, and 0. 29 ( ×10^6 L/mol), respectively. The stoichiometry of the metal ions binding to PC was determined using the method of continuous variations. Fluorescence quenching titration with metal ions have revealed that the stoichiometry of Al^3+, Cu^2+, Sn^2+, Zn^2+-bound PC were 1:2, 1:2, 1:1 and 1:2, respectively. These quantitative models of PC-metal interaction will improve the understanding of the likely physiological function of PC.

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