Species-selective analysis for metal-biomacromolecular complexes using hyphenated techniques

Analytical chemistry of metal complexes with biomacromolecules based on the coupling of a high resolution separation technique with an element or species selective detection technique is critically discussed. The role of size-exclusion chromatography (SEC) with on-line atomic spectrometric detection is evaluated for the characterization of the metal distribution among the fractions of different molecular weight. Attention is given to the conditions for the separation of metallated biomacromolecular isoforms and subisoforms by anion-exchange and reversed-phase HPLC. Techniques for interfacing chromatography with atomic absorption spectrometry (AAS), inductively coupled plasma atomic emission spectrometry (ICP AES) and ICP mass spectrometry (ICP MS) are assessed. The potential of electrospray (tandem) mass spectrometry for the on-line determination of the molecular mass of the eluting protein is highlighted. Perspectives for capillary zone electrophoresis (CZE), microbore and capillary HPLC with ICP MS and electrospray MS detection for probing metalloproteins are discussed. Applications of hyphenated techniques to the analysis of real-world samples are reviewed.