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

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H_(2)O_(2)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H_(2)O_(2)浓度:0.025 mol/L、hemin-ficin浓度2.0 mmol/L和孵育时间10s。抗坏血酸浓度在0.005~0.500mmol/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(r^(2)=0.9912),检出限为0.067μmol/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。

Objective To establish a rapid system for the colorimetric detection of ascorbic acid based on hemin-ficin complex peroxidase-like.Methods The effects of H_(2)O_(2)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:H_(2)O_(2)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(r^(2)=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.