Separation of Lipoproteins From Human Plasma by Flow Field-Flow Fractionation

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Abstract
A flow field-flow fractionation (flow FFF) system was used in both isocratic and programmed-field procedures to rapidly analyze and characterize the HDL, LDL, and VLDL fractions of human blood plasma. In this paper, the general principles and theory of separation are briefly reviewed. The theoretically predicted retention values are shown to compare favorably with the experimental results. The sample recovery and system reproducibility were determined. The lipoprotein fractions were clearly separated into different peaks, although the peaks tended to be rather broad, predominantly due to the sample polydispersity and, to a smaller extent, due to systemic bandbroadening. Plasma samples were analyzed without sample pre-treatment and differences in lipoprotein profiles were observed for different individuals. Not only could the HDL, LDL, and VLDL fractions be separated, but lipoprotein subspecies were also determined with the use of a programmed field. The hydrodynamic sizes and diffusion coefficients of plasma lipoproteins were deduced from their retention behavior based on FFF theory. The characterization of lipoprotein fractions, based on size or diffusion coefficient, provided additional information which may be useful for research or diagnostic purposes.