Metal ion complexes of EDTA: a solute system for density gradient ultracentrifugation analysis of lipoproteins.

In the study reported here, we apply some of the features of coordination chemistry to solve a long-standing problem in the separation and characterization of lipoprotein particles. Lipoproteins are circulating micelle-like particles responsible for lipid transport. They exist in three major classes: very-low-density lipoprotein, low-density lipoprotein, and high-density lipoprotein in well-defined density ranges using the density gradient ultracentrifugation (DGU) method. The analytical instrumentation of DGU has improved over the years in response to clinical evidence that certain lipoprotein species are linked to a high risk for developing cardiovascular disease. A long-standing problem has been a lack of appropriate gradient-forming solutes that can generate a useful gradient from a homogeneous solution. We have found that a new class of solutes based on metal ion complexes has the potential of providing a wide selection of compounds where the features can be modulated by choice of ligand, complexing metal ion, and counterion. In this study, we have chosen the cesium salt of BiEDTA (CsBiEDTA) and have investigated the dynamics of density gradient formation in the ultracentrifuge. We show that a useful density gradient can be formed within a few hours beginning with a homogeneous solution. We also present data on the migration behavior of lipoproteins under gradient-forming conditions and show that high-resolution density profiles can be obtained with good precision. The resolution of the CsBiEDTA profile is compared with those obtained using high molecular weight organic solutes.