摘要
使用Superdex 75高效凝胶过滤柱的高效液相色谱法分离并测定了大鼠肝中的金属硫蛋白。在测定前,先将肝匀浆上清液进行镉饱和以及加热的简单前处理。大鼠肝中的金属硫蛋白在短时间内被彻底分离,并在250nm的紫外吸收下直接定量测定。本方法简便易行、精密度好(CV=1.4%,n=10)、回收率高(90%以上)。金属硫蛋白的检测限为0.265μg。本方法与镉饱和法作对照,显示有良好的相关关系(r=0.99)。
Metallothionein(MT)in rat liver was isolated and determined by high-per- formance liquid chromatography(HPLC)with a Superdex 75 high performance gel filtration column.The liver supernatant was pretreated by cadmium saturation and heating before HPLC determination.MT in rat liver was completely separated in a short time,and was quantitated directly as a function of UV absorbance at 250 nm.The quantitation was simpli- fied by using a commercially available horsekidney MT standard.The method has a good precision(CV=1.4%,n=10)and a good recovery(exceed 90%).The detection limit for MT was 0.265μg.The method was compared with the traditional cadmium saturation method fordetermination of hepatic MT of rat and there was good correlation between these two methods(r=0.99).
关键词
金属硫蛋白
肝
高效液相色谱法
大鼠
metallothionein
liver
high-performance liquid chromatography
rat
