京尼平苷为底物测定β-葡萄糖苷酶活力的方法

Method for Determination of β- glucosidase Activity for Geniposide

研究以京尼平苷为底物氨基酸为显色剂测定β-葡萄糖苷酶活力的方法。β-葡萄糖苷酶水解京尼平苷的温度为50℃,pH为5.0,京尼平苷浓度为0.625mmol/L,水解10min后,立即加入1ml1mol/LNa2CO3终止反应,混匀,再加入体积比为1:1的0.2mg/ml的精氨酸溶液,沸水浴显色10min,冷却后于590nm处测光吸收度值。该方法的检测线性范围为0.05～1U/ml,相关系数为0.9998,检测限为0.02U/ml,精密度为1.5%(n=5),回收率为99.5%～101.1%,该方法准确度高,结果稳定。

A method for determination of β- glucosidase activity using geniposide as substrate by amino acid colorimetry was reported. The hydrolysis conditions of geniposide with β - glucosidase and the color reaction conditions, were optimized as： hydrolyzing temperature 50℃, time 10min, pH5.0, color reaction temperature 100℃, color reaction time 10min with the volume ratio of geniposide solvent to Arg solvent 1：1.The method measurement range is 0.05 - 1U/ml, the linear correlationship 0.9998, the detection limit 0.02U/ml and RSD 1.5%（n=5） with the recovery rate is 99.5% to 101. 1%.This method has the advantages of wide measuring range, and the determination of β- glucosidase activity.