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Determination and purification of metallothioneins by high performance liquid chromatography
Author(s) -
Pan Aihua,
Wang Zhenxin,
Ru Binggen
Publication year - 1991
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130050503
Subject(s) - chemistry , metallothionein , chromatography , high performance liquid chromatography , elution , cadmium , detection limit , gradient elution , organic chemistry
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 60 Co‐irradiated samples. A detection limit of 5 μg/g liver was established for this method. We have analysed more than 100 biological samples and obtained satisfactory results.