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Journal of Lipid Research, Vol. 22, 1015-1020, August 1981
Copyright © 1981 by Lipid Research, Inc.

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

G. Russell Warnick ¹, John J. Albers ¹, Paul S. Bachorik ², John D. Turner ³, Carmalita Garcia ⁴, Carl Breckinridge ⁵, Kanta Kuba ⁶, Susan McNeely ⁷, Gerald Hillerman ⁸, Paul King ⁹, Richard Muesing ¹⁰, Bernard Most ¹¹, and Ken Lippel ¹²

¹ Northwest Lipid Research Clinic, Harborview Medical Center, Seattle, WA 98104 and Department of Medicine, University of Washington School of Medicine, Seattle, WA 98195.
² The John Hopkins University, Baltimore, MD 21205.
³ Lipid Research Clinic, University of California, San Diego, La Jolla, CA 92093.
⁴ Stanford University School of Medicine, Stanford, CA 94305.
⁵ Lipid Research Clinic, Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8.
⁶ Department of Biochemistry, University of Minnesota School of Medicine, Minneapolis, MN 55455.
⁷ University of Cincinnati Medical Center, Department of Internal Medicine, College of Medicine, Cincinnati, OH 45229.
⁸ Clinical Analytical Laboratory, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104.
⁹ Department of Internal Medicine, Iowa Lipid Research Clinic, University of Iowa College of Medicine, Iowa City, IA 52242.
¹⁰ George Washington University, Lipid Research Laboratory, Washington, DC 20037.
¹¹ University of North Carolina, Chapel Hill, NC 27514.
¹² National Institutes of Health, National Heart, Lung, and Blood Institute, Lipid Metabolism Branch, Bethesda, MD 20205.

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 Mn²⁺. 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-Mn²⁺ 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-Mn²⁺ supernates is a convenient alternative to precipitation after either dilution or removal of VLDL.—Warnick, G. R., J. J. Albers, P. Bachorik, J. Turner, C. Garcia, C. Breckinridge, K. Kuba, S. McNeely, G. Hillerman, P. King, R. Muesing, B. Most, and K. Lippel. Multilaboratory evaluation of an ultrafiltration procedure for high density lipoprotein cholesterol quantification in turbid heparin-manganese supernates.

Supplementary key words coronary heart disease • lipoprotein quantitation • immunoassay

Submitted on November 6, 1980
Revised on April 27, 1981

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				G. R. Warnick, M. Nauck, and N. Rifai Evolution of Methods for Measurement of HDL-Cholesterol: From Ultracentrifugation to Homogeneous Assays Clin. Chem., September 1, 2001; 47(9): 1579 - 1596. [Abstract] [Full Text] [PDF]