Determination and purification of metallothioneins by high performance liquid chromatography.
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.
Pan
A H
AH
Department of Biology, Peking University, Beijing, China.
Wang
Z X
ZX
Ru
B G
BG
D000818:Animals; D002104:Cadmium; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000737:chemistry; D008668:Metallothionein / Q000032:analysis / Q000302:isolation & purification*; D051379:Mice; D011817:Rabbits
D002104:Cadmium; D008668:Metallothionein