title: Determination and purification of metallothioneins by high performance liquid chromatography.
abstract: 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.
authors.affiliation: Department of Biology, Peking University, Beijing, China.
authors.forename: A H
authors.initials: AH
authors.lastname: Pan
authors.forename: Z X
authors.initials: ZX
authors.lastname: Wang
authors.forename: B G
authors.initials: BG
authors.lastname: Ru
mesh_terms: 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
chemical_list: D002104:Cadmium; D008668:Metallothionein