孔雀绿比色法测定5′-核苷酸酶的活性  

Determination of 5′-nucleotidase by Malachite green colorimetric method

作　　者：左凤琼[1] 李虹[1] 吴艳红[1] 吴兆锋[1] 邓如伟[2] 李颖[2] 李明远[1] 

机构地区：[1]四川大学华西基础医学与法医学院,四川成都610041 [2]四川大学华西药学院,四川成都610041

出　　处：《华西药学杂志》2004年第6期435-436,共2页West China Journal of Pharmaceutical Sciences

摘　　要：目的　建立测定细胞膜标志酶活性的方法。方法　采用孔雀绿比色法测定。结果　孔雀绿比色法可测出 0 1μg的磷 ,重复性好 ,RSD =1 95 % ,回收率为 97 5 2 % ,显色稳定 ,操作简便 ,可灵敏地反映 5′ -核苷酸酶的活性。 5′ -核苷酸酶的活性的高低可反映细胞膜的纯度。结论　通过对酶反应时间和浓度以及重复性和回收率的考察 ,建立了检测 5′ -核苷酸酶的最佳条件。OBJECTIVE To establish a new method for the determination of 5′-nucleotidase(5-NT). METHODS Activity of 5′-nucleotidase is reflected by the content of phosphor determined by Malachite green colorimetric(MGC) method . RESULTS 0.1 μg of phosphor could be determined by MGC method with RSD of 1.95% and the recovery of 97.52%. MGC assay could sensitively determine 5′-NT and activity can reflect the purity of cell membrane.CONCLUSION The variance coefficient, recovery, reaction time and concentration of the enzyme have been studies. The best method to detemine 5′-nucleotidase are established.

关 键 词：核苷酸 孔雀绿 比色法测定 酶活性 细胞膜 RSD 灵敏 浓度 纯度 回收率 

分 类 号：R931.3[医药卫生—生药学] R383.33[医药卫生—药学]