坚果中还原型谷胱甘肽（GSH）和蛋白巯基（P-SH）含量的测定

发布时间：2018-04-18 22:29

本文选题：谷胱甘肽 + 巯基化合物　；参考：《延边大学》2017年硕士论文

【摘要】：绪论概述了坚果中含有的营养成分和现阶段对坚果的研究情况,介绍了细胞内巯基化合物在生物体内的重要作用及其常用的分析检测方法,并提出坚果中谷胱甘肽等含半胱氨酸残基的巯基化合物的测定方法及其研究的重要意义,概述了本研究的选题思路和主要研究内容。HPLC法测定四种食用坚果种子中还原型谷胱甘肽(GSH)和蛋白巯基(P-SH)含量用5,5'-二硫双-2-硝基苯甲酸(DTNB)为柱前衍生化试剂,在细胞超声破壁的同时立即与衍生化试剂反应,做到同步提取,HPLC梯度洗脱法测定了开心果、巴旦木、澳洲坚果、腰果干品中谷胱甘肽(GSH)和含半胱氨酸残基的蛋白巯基(P-SH)含量。结果发现四种坚果均含较丰富的GSH和P-SH巯基,其含量分别为,开心果:GSH=1.2303 μmol/g,P-SH=3.5414 μmol/g,巴旦木:GSH=0.3069μmol/g,P-SH=1.9541 μmol/g,澳洲坚果:GSH=1.5708 μmol/g,P-SH=0.5905 μmol/g,腰果:GSH=0.3373 μmol/g,P-SH=2.2458 μmol/g。在优化的选定条件下,测定方法的回收率为99.37%～101.45%,GSH和P-SH测定检测限分别为0.1386 μmol/g和0.2184 μmol/g。本研究对坚果活性成分的构成及其巯基化合物在保健防病作用研究及其功能性食品开发均提供了有用的参考数据。光谱法测定四种食用坚果种子中谷胱甘肽(GSH)和总巯基(-SH)的含量利用邻苯二甲醛(OPA)和5,5'-二硫双-2-硝基苯甲酸(DTNB)为衍生化试剂,通过荧光光谱法和UV-vis吸收光谱法,测定了坚果中还原型谷胱甘肽(GSH)和总巯基(-SH)的含量。测定结果表明:坚果中富含GSH和总巯基,开心果GSH=1.0847 μmol/g、-SH=5.0001 lμmol/g 澳洲坚果 GSH=1.4720 μmol/g、-SH=2.4774μmol/g,腰果 GSH=0.3255 μmol/g、-SH=2.9302 μmol/g,巴旦木 GSH=0.2890 μmol/g、-SH=2.7627 μmol/g。回收率范围为 99.72%～100.90%和 98.66%～100.75%。坚果中可能的共存组分对测定的影响探究了坚果中的维生素、氨基酸、不饱和脂肪酸等其他活性营养物质对本研究中还原型谷胱甘肽(GSH)和总巯基(-SH)测定的影响。测定结果表明在光谱法测定中较高浓度的维生素和不饱和脂肪酸可能会对结果产生一定的影响,导致总巯基的含量测定结果偏高,而色谱法测定中加入的干扰物质对谷胱甘肽和总巯基含量的测定基本无影响。
[Abstract]:The introduction summarizes the nutritional components in nuts and the current research situation of nuts, and introduces the important role of intracellular sulfhydryl compounds in organisms and the commonly used analytical methods.The method for the determination of thiol compounds containing cysteine residues in nuts and the significance of their research are put forward.The main contents of this study. Determination of reduced glutathione glutathione (GSH) and protein mercapto P-SH) in four kinds of edible nut seeds by HPLC was studied. DTNB was used as precolumn derivatization reagent.The contents of glutathione glutathione (GSH) and cysteine residue protein (P-SHH) in pistachios, alendanthus, macadamia nuts and cashew nuts were determined by HPLC gradient elution method.The results showed that the four kinds of nuts were rich in GSH and P-SH sulfhydryl groups, and the contents of them were: pistachio: GSH 1.2303 渭 mol / g, P -SHN 3.5414 渭 mol / g, Dendanth: 1. 3069 渭 mol / g P -SHN 1.9541 渭 mol / g, Macadamia nut 1. 5708 渭 mol / ggP -SHN 0. 5905 渭 mol / g, cashew w / w GSH 0. 3373 渭 mol / g P -SHU 2.2458 渭 mol / g / g.Under the optimized conditions, the detection limits of GSH and P-SH were 0.1386 渭 mol/g and 0.2184 渭 mol / g, respectively.This study provides useful reference data for the study of the composition of the active components of nuts, the effects of sulfhydryl compounds on health and disease prevention and the development of functional foods.Spectrophotometric determination of glutathione (GSH) and total mercapto (SH) in four kinds of edible nut seeds using phthalaldehyde (OPA) and 5-dithiobis dithiobenzene (DTNB) as derivation reagents, fluorescence spectrometry and UV-vis absorption spectrometry were used to determine the content of glutathione (GSH) and total mercapto (-SH) in edible nut seeds.The contents of reduced glutathione glutathione (GSH) and total mercapto (SH) in nuts were determined.The results showed that the nuts were rich in GSH and total sulfhydryl, pistachio GSH=1.0847 渭 mol / g ~ (-1) -SHN 5.0001 渭 mol/g, macadamia nut GSH=1.4720 渭 mol / g ~ (-1) / g ~ (-1) / h ~ (-1) 2.4774 渭 mol / g, cashew GSH=0.3255 渭 mol / g ~ (-1) -SH ~ (2 +) 2.9302 渭 mol / g 路g ~ (-1), GSH=0.2890 渭 mol 路g ~ (-1) -SHN 2.7627 渭 mol / g ~ (-1) 路g ~ (-1) 路g ~ (-1).The recoveries ranged from 99.72% to 100.90% and 98.66 to 100.75%.Effects of possible coexisting components in nuts on the determination of glutathione glutathione (GSH) and total mercapto (SH) in nuts the effects of other active nutrients such as vitamins amino acids and unsaturated fatty acids on the determination of GSH were investigated.The results showed that the higher concentration of vitamins and unsaturated fatty acids in the determination of total sulfhydryl group might have a certain effect on the results, which resulted in the higher content of total sulfhydryl group.However, the content of glutathione and total mercapto was not affected by the addition of interfering substances in chromatographic determination.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS255.6;O652

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