Complications in the determination of HDL2/HDL3 ratios

We investigated different procedures to subfractionate high density lipoprotein (HDL) into its density subfractions (HDL2 and HDL3). When pore-limit as well as zone polyacrylamide gel electrophoresis was used, it proved to be necessary to use freshly prepared and plasma protein-free HDL preparations. For a quantitative subractionation of HDL in HDL2 and HDL3 we also tested density gradient ultracentrifugation. To avoid interference with LDL contamination a low starting density was required. Both methods, electrophoresis as well as density gradient ultracentrifugation were too laborious and time-consuming for large scale use. We devised a combination of both ultracentrifugation and electrophoresis as a method which is relatively appropriate for determination of HDL2/HDL3 ratios on large scale.