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Metal Ion Complexes of EDTA:  A Solute System for Density Gradient Ultracentrifugation Analysis of Lipoproteins

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Department of Chemistry, Texas A&M University, College Station, Texas 77843
Cite this: Anal. Chem. 2005, 77, 1, 200–207
Publication Date (Web):December 2, 2004
https://doi.org/10.1021/ac0490402
Copyright © 2005 American Chemical Society
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Abstract

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.

*

 To whom correspondence should be addressed. E-mail:  [email protected] mail.chem.tamu.edu. Phone:  (979)-845-2021. Fax:  (979)-845-8987.

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  4. R. K. Mudsainiyan, S. K. Chawla. Synthesis, Characterizations, Crystal Structure Determination of μ 6 Coordinated Complex of Co (III) with EDTA and Its Thermal Properties. Molecular Crystals and Liquid Crystals 2015, 606 (1) , 237-245. https://doi.org/10.1080/15421406.2014.916531
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  17. Richa Chandra, Ronald D. Macfarlane. Remnant Lipoprotein Density Profiling by CsBiEDTA Density Gradient Ultracentrifugation. Analytical Chemistry 2006, 78 (3) , 680-685. https://doi.org/10.1021/ac050775w
  18. I. Leticia Espinosa, Catherine J. McNeal, Ronald D. Macfarlane. Method for Lipoprotein(a) Density Profiling by BiEDTA Differential Density Lipoprotein Ultracentrifugation. Analytical Chemistry 2006, 78 (2) , 438-444. https://doi.org/10.1021/ac050962u
  19. Jeffery D. Johnson, Natalie J. Bell, Erin L. Donahoe, Ronald D. Macfarlane. Metal Ion Complexes of EDTA as Solutes for Density Gradient Ultracentrifugation:  Influence of Metal Ions. Analytical Chemistry 2005, 77 (21) , 7054-7061. https://doi.org/10.1021/ac0509657
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