Multilaboratory evaluation of an ultrafiltration procedure for high density lipoprotein cholesterol quantification in turbid heparin-manganese supernates.

Abstract

High density lipoprotein (HDL) can be quantitated by measurement of cholesterol in supernates after precipitation of low and very low density lipoprotein (LDL and VLDL) with heparin and Mn(2+). Supernatant turbidity, often observed with hypertriglyceridemic specimens, indicates incomplete sedimentation of LDL/VLDL and precludes accurate quantitation of HDL. Ten Lipid Research Clinic Laboratories compared an ultrafiltration technique for clearing turbid heparin-Mn(2+) supernates to current methods involving repeat precipitation of either the original specimen after dilution or the d > 1.006 g/ml fraction after removal of VLDL from the initial specimen by ultracentrifugation. Results for ultrafiltration of 429 turbid supernates averaged only slightly higher (1.0-1.1 mg/dl) than results by the dilution or ultracentrifugation methods on the same specimens, but this difference was found to be significant (P < 0.005). The agreement of the ultrafiltration method with the other two methods is indicated by the following linear regression equations: a), ultrafiltration = (0.964 x ultracentrifugation) + 2.4 mg/dl, and correlation coefficient = 0.926; and b), ultrafiltration = (0.936 x dilution) + 3.3 mg/dl, and correlation coefficient = 0.933. We conclude that ultrafiltration of turbid heparin-Mn(2+) supernates is a convenient alternative to precipitation after either dilution or removal of VLDL.