基于氯化血红素-无花果蛋白酶复合物比色检测抗坏血酸的研究

Study on colorimetric determination of ascorbic acid based on the hemin-ficin complex

中文关键词:  无花果蛋白酶  氯化血红素  过氧化物模拟酶  抗坏血酸  比色检测

英文关键词:ficin  hemin  peroxidase-like  ascorbic acid  colorimetric

中文摘要:

      目的 利用氯化血红素-无花果蛋白酶复合物(hemin-ficin)模拟过氧化物酶活性构建抗坏血酸快速比色检测体系。方法 通过单因素实验优化H2O2浓度、hemin-ficin浓度和孵育时间3个因素对hemin-ficin复合物比色检测体系的影响, 利用正交实验设计优化获得最佳检测条件。同时对体系的灵敏度、抗干扰性和稳定性进行评估。结果 hemin-ficin复合物催化体系的最优条件为H2O2浓度: 0.025 mol/L、hemin-ficin浓度2.0 mmol/L和孵育时间10 s。抗坏血酸浓度在0.005~0.500 mmol/L范围时, 吸光度差值与抗坏血酸浓度的工作曲线具有良好的线性关系(r2=0.9912), 检出限为0.067 μmol/L, 实际样品加标回收率为97.45%~102.11%, 相对标准偏差低于2.45%。结论 基于hemin-ficin复合物模拟酶活性所建立的抗坏血酸比色检测体系, 具有极高的灵敏度、良好的抗干扰性和稳定性, 该研究成果将为抗坏血酸等食品成分的分析检测提供一条新的思路。

英文摘要:

      Objective To establish a rapid system for the colorimetric detection of ascorbic acid based on hemin-ficin complex peroxidase-like. Methods The effects of H2O2 concentration, hemin-ficin concentration and incubation time on the colorimetric detection system of hemin-ficin complex were optimized by single factor experiment, and the optimal detection conditions were obtained by orthogonal experimental design optimization. At the same time, the sensitivity, anti-interference and stability of the system were evaluated. Results The optimum detection conditions of hemin-ficin complex catalytic system were as follows: H2O2 concentration of 0.025 mol/L, hemin-ficin concentration of 2.0 mmol/L and incubation time of 10 s. When the concentration of ascorbic acid was in the range of 0.005?0.500 mmol/L, the working curve between the absorbance difference and the concentration of ascorbic acid had a good linear relationship (r2=0.9912), the limit of detection was 0.067 μmol/L, and the recoveries of the actual samples were 97.45%?102.11% with a relative standard deviation below 2.45%. Conclusion The ascorbic acid colorimetric detection system based on the simulated enzyme activity of hemin-ficin complex has extremely high sensitivity, good anti-interference and stability, the research results will provide a new idea for the analysis and detection of ascorbic acid and other food components.