SEPARATION OF METALLOPROTEIN COMPLEXES IN SERUM BY SIZE-EXCLUSION CHROMATOGRAPHY - OPTIMIZATION OF THE SEPARATION PARAMETERS RETENTION BEHAVIOR AND RECOVERY EMPLOYING RADIOTRACERS

A suitable procedure was developed for speciation analysis of metallop rotein complexes in serum using directly coupled size-exclusion chroma tography and an element-specific detector. Two column matrices used fo r size-exclusion chromatography (TSK G 3000 SW and Asahipak GS 520) we re investigated with respect to the recovery and retention behaviour f or metalloprotein complexes. Optimisation of the separation parameters (buffer type, concentration, pH) was achieved by means of metalloprot ein complexes marked with radiotracers. For speciation of serum the ma trix in the Asahipak GS column is more efficient. Given optimal eluent characteristics (100 mM Tris, pH 7.4) the recovery of the elements in vestigated (sodium, calcium, iron and zinc) was 100%. Further, the ret ention behaviour (retention time, ratios of the peak areas) remained u nchanged for several successive separations. (C) 1998 Elsevier Science B.V.