Pan A H, Wang Z X, Ru B G
Department of Biology, Peking University, Beijing, China.
Biomed Chromatogr. 1991 Sep;5(5):193-7. doi: 10.1002/bmc.1130050503.
A rapid, reproducible and sensitive high performance liquid chromatography (HPLC) method for the determination and purification of metallothionein-I (MT-I) and metallothionein-II (MT-II) in mouse and rabbit livers has been developed. Methallothioneins (MTs) were separated by an HPLC anion exchange column, eluted through a linear gradient of Tris buffer and the peak containing MTs was determined by atomic absorption spectrophotometry. Furthermore, the content of MT-I or MT-II was calculated by protein peak area in a short time (about 20 min). The sample to be tested was homogenized, centrifuged and saturated by cadmium. MT-I and MT-II were eluted at 15.9 and 19.3 min, respectively. The following mouse liver cytosols were tested: controls, Cd-injected samples and 60Co-irradiated samples. A detection limit of 5 micrograms/g liver was established for this method. We have analysed more than 100 biological samples and obtained satisfactory results.
已开发出一种快速、可重复且灵敏的高效液相色谱（HPLC）方法，用于测定和纯化小鼠和兔肝脏中的金属硫蛋白-I（MT-I）和金属硫蛋白-II（MT-II）。金属硫蛋白（MTs）通过HPLC阴离子交换柱分离，用Tris缓冲液线性梯度洗脱，并通过原子吸收分光光度法测定含MTs的峰。此外，MT-I或MT-II的含量可在短时间内（约20分钟）通过蛋白质峰面积计算得出。待测样品经匀浆、离心并用镉饱和。MT-I和MT-II分别在15.9分钟和19.3分钟洗脱。对以下小鼠肝脏胞质溶胶进行了检测：对照组、注射镉的样品和60Co照射的样品。该方法的检测限为5微克/克肝脏。我们已经分析了100多个生物样品并获得了满意的结果。
