Precipitation with polyethylene glycol and density-gradient ultracentrifugation compared for determining high-density...

The purpose of this study was to compare quantification of cholesterol in high-density lipoprotein subfractions HDL2 and HDL3 by precipitation with polyethylene glycol (PEG) with that by density-gradient ultracentrifugation. Fresh serum samples from 32 fasting, obese children were analyzed with precipitation reagent "Quantolip" (Immuno AG), and then fractionated with a Beckman TL 100 ultracentrifuge with a swinging-bucket rotor. After centrifugation we carefully removed the supernate with a syringe and measured the cholesterol from each fraction enzymatically with CHOD-PAP reagent (Boehringer Mannheim). The low-density lipoprotein (LDL-), HDL2-, and HDL3-cholesterol values measured by ultracentrifugation did not differ significantly from those obtained by precipitation; the correlation coefficients (r) between the two methods were 0.96 for LDL, 0.75 for HDL2, and 0.96 for HDL3. The relatively simple PEG precipitation method used in this study measures total HDL and its major subclasses HDL2 and HDL3 with accuracy and precision comparable with those of the well-established ultracentrifugation method.