Heparin--Mn2+ quantitation of high-density-lipoprotein cholesterol: an ultrafiltration procedure for lipemic samples

We describe a modified heparin--Mn2+ procedure for high-density-lipoprotein cholesterol quantitation, especially in lipemic samples. High-density-lipoproteins may be estimated as cholesterol remaining in plasma supernates after precipitation of other lipoproteins by heparin and Mn2+ treatment. However, in lipemic samples or those from non-fasting individuals, the lower density of the precipitated chylomicrons, very-low-, and low-density-lipoproteins frequently prevents their sedimentation by the usual low-speed centrifugation, and high-density-lipoprotein cholesterol thus is overestimated in the resulting turbid supernates. Sedimentation is improved by a twofold increase in Mn2+ concentration to 92 mmol/liter. The procedure reported here produced clear supernates in more than 95% of samples tested. Any remaining turbid supernates can be cleared by a simple, convenient ultrafiltration technique. The filtration removed essentially all of the very-low- and low-density-lipoproteins without removing appreciable amounts of high-density-lipoproteins.