1. 西北农林科技大学食品科学与工程学院
2. 陕西杨凌,712100
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杨阳, 刘光勤, 杨思龙, 等. 基于Ce-BDC的氧化物酶活性检测果汁中抗坏血酸的比色方法研究[J]. 分析测试学报, 2021,40(5):678-683.
Study on Colorimetric Detection of Ascorbic Acid in Juice Based on Ce-BDC Oxidase like Activity[J]. 2021,40(5):678-683.
以对苯二甲酸(H2BDC)为有机配体,Ce为活性金属中心制备了一种具有氧化物酶活性的金属有机框架材料Ce-BDC,其可以催化无色3,3,5,5-四甲基联苯胺(TMB)生成在652 nm处具有强紫外吸收峰的蓝色氧化反应产物oxTMB。该氧化物模拟酶的催化活性受到pH值和温度的影响,其催化规律遵循米氏反应动力学。抗坏血酸的加入可将oxTMB还原导致溶液蓝色减弱且652 nm处的吸光度下降,该文基于这一现象建立了一种果汁中检测抗坏血酸的比色方法。在优化条件下,抗坏血酸浓度在1.0~30 μmol/L范围内与其652 nm处的吸光度呈良好线性,相关系数为0.991 4,检出限为0.32 μmol/L。方法对实际果汁样品中抗坏血酸的回收率为98.2%~102%,相对标准偏差(RSD)为1.2%~7.2%。该策略可为基于纳米酶的比色方法检测抗坏血酸提供新思路。
Ascorbic acid(AA),also known as vitamin C(Vc),is a type of reducing substance with an enediol structure.Ascorbic acid plays an important role in maintaining the normal physiological functions of the human body.Appropriate intake of ascorbic acid is beneficial to enhance the bodys anti-stress ability and immunity,promote the absorption of calcium,iron,folic acid,etc.,and prevent cancer and cardiovascular disease.In addition,ascorbic acid as an antioxidant is also widely used in the fields of medical pharmacy,food processing and cosmetic products.Therefore,it is very necessary to develop an accurate and sensitive method for ascorbic acid detection.Colorimetric analysis methods based on nanozymes have attracted more and more research interest owing to their fascinating advantages of low cost,good stability,high sensitivity and excellent selectivity.In this paper,a simple,sensitive,rapid and selective colorimetric method for ascorbic acid detection was developed.Inspired by the fact that nanozymes with redox-active metal center are prone to possess good catalytic performance,a novel metal-organic framework nanozyme Ce-BDC was first prepared by using terephthalic acid (H2BDC) as organic ligand and Ce as active metal center.The as-prepared Ce-BDC was well characterized by scanning electron microscopy(SEM),X-ray diffraction(XRD) and X-ray photoelectron spectroscopy(XPS).Then the oxidase-like activity of Ce-BDC was revealed and Ce-BDC could catalyze the colorless 3,3,5,5-tetramethylbenzidine(TMB) to produce a deep blue oxidation product oxTMB which has a strong UV absorption peak at 652 nm.The catalytic activity of Ce-BDC follows the Michaelis-Menten kinetics,which could be affected by pH value and temperature condition.Results showed that the optimal reaction temperature is 30 ℃,the optimal reaction pH value is 3.0,and the optimal reaction time is 120 s.Due to the strong reducing property of ascorbic acid,the addition of ascorbic acid could reduce oxTMB into colourless TMB,resulting in the fading of blue color and the decrease of absorbance at 652 nm.Based on this phenomenon,a colorimetric method was established for ascorbic acid detection.Under the optimum reaction conditions,the absorbance of the reaction solution at 652 nm has a good linear relationship with the concentration of ascorbic acid in the range of 1.0-30 μmol/L,and the detection limit for ascorbic acid was found to be 0.32 μmol/L,which could satisfy the requirements for the practical detection of ascorbic acid.The established method was applied in the detection of ascorbic acid in fruit juice sample via standard addition test.The recovery for ascorbic acid was in the range of 98.2%-102%,with the relative standard deviations of 1.2%-7.2%,demonstrating that the method is promising for the rapid and selective detection of ascorbic acid.In a word,this work provides a new approach for the colorimetric detection of ascorbic acid in fruit juice based on nanozyme.
抗坏血酸金属有机框架模拟氧化物酶活性比色检测
ascorbic acidmetal-organic frameworkoxidase-like activitycolorimetric detection
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