Gel permeation, ion-exchange and reversed-phase columns for separation of metallothioneins by high-performance liquid chromatography-atomic absorption spectrophotometry.
Metallothioneins were separated on three kinds of HPLC column that differ in separation principle: gel permeation, ion exchange and reversed phase. Elution of the columns at neutral to weakly basic buffer conditions prevented dissociation of the metals, and the metals bound to metallothioneins were quantitatively eluted out. The metals in the eluate were continuously determined on an atomic absorption spectrophotometer by introducing the eluate directly to the nebulizer tube. Prerequisites for the on-line separation-detection system are discussed.
D000818:Animals; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D008099:Liver / Q000032:analysis; D008668:Metallothionein / Q000302:isolation & purification*; D051381:Rats; D013054:Spectrophotometry, Atomic / Q000379:methods; D014982:Xenopus laevis
D008668:Metallothionein