透明质酸酶活性抑制实验的影响因素研究

Study of factors influencing hyaluronidase activity inhibition experiments

以研究透明质酸酶活性抑制实验影响因素为目的,基于Elson-Morgan改良法反应原理,以吸光度为指标,通过单因素实验,研究了显色时间、底物水解时间以及底物浓度对透明质酸酶活性抑制实验的影响。结果表明:当显色时间为20min、底物水解时间为40min时,显色反应的吸光度趋于稳定;底物浓度为1.2mg/mL时,显色反应吸光度最优。分析了甘草酸二钾抑制透明质酸酶活性抑制率的重现性实验,相对标准偏差为1.66%,表明实验确定因素重现性好。考察了甘草酸二钾透明质酸酶抑制率的线性关系,并根据线性方程计算出甘草酸二钾的半抑制浓度(IC50值)为0.18mg/mL。

The effect of color development time,substrate hydrolysis time and substrate concentration on the inhibition of hyaluronidase activity by single-factor experiment was investigated,based on the reaction principle of Elson-Morgan modified method with absorbance as the index.The absorbance of the color development reaction was stabilized when the color development time was 20min and the substrate hydrolysis time was 40min,and the absorbance of the color development reaction was optimal when the substrate concentration was 1.2mg/mL.The reproducibility experiment of the inhibition rate of hyaluronidase activity inhibition by dipotassium glycyrrhizinate was analyzed,and the relative standard deviation was 1.66%,which indicated that the reproducibility of the determining factor of the test was good.The linearity of dipotassium glycyrrhizate hyaluronidase inhibition was examined,and the ICso value of dipotassium glycyrrhizate was calculated as 0.18mg/mL according to the linear equation.