牛血清白蛋白荧光猝灭法同时测定2种细胞分裂素

Simultaneous measurement of two cytokinins by bovine serum albumin fluorescence quenching method

基于牛血清白蛋白(BSA)荧光偏振和荧光猝灭信号建立了区分及同时测定6-苄嘌呤(6-BA)和激动素(KT)的方法。在中性条件下,牛血清白蛋白与6-BA和KT发生疏水和氢键等作用并导致牛血清白蛋白荧光信号猝灭强度的不同,利用荧光偏振猝灭信号计算得到荧光偏振度(p)和荧光各向异性(r),BSA-(6-BA)体系对应较大的荧光偏振度(p)和荧光各向异性(r),而BSA-KT体系对应较小的P和r,利用两体系P和r数值上的差距可达到区分6-BA和KT的目的。利用荧光猝灭法和双标准曲线计量分析法实现了对市售苦瓜和番茄样品表皮中6-BA和KT的同时测定,效果令人满意。

Based on the fluorescence quenching signal of bovine serum albumin( BSA),a new method was established to distinguish and simultaneously measure 6-benzylaminourine( 6-BA) and kinetin( KT). Under neutral conditions,BSA could react with 6-BA and KT by hydrophobic and hydrogen bonding effects,resulting in different quenching intensity of BSA fluorescence signal. The degree of fluorescence polarization( p) and fluorescence anisotropy( r) were calculated by using fluorescence quenching signal. The p and r of BSA-( 6-BA)were larger than those of BSA-KT system,and the difference between the p and r values of the two systems could be used to distinguish 6-BA and KT. Then the simultaneous measurement of 6-BA and KT in the skins of commercially available balsam pear and tomato samples was achieved by the double standard curve measurement analysis method and the results were satisfactory.