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Journal of Lipid Research, Vol. 19, 65-76, January 1978
Copyright © 1978 by Lipid Research, Inc.
A comprehensive evaluation of the heparin - manganese precipitation procedure for estimating high density lipoprotein cholesterol
G. Russell Warnick and John J. Albers
Northwest Lipid Research Clinic, Harborview Medical Center, University of Washington School of Medicine, Seattle, WA 98104
The accurate quantitation of high density lipoproteins has recently assumed greater importance in view of studies suggesting their negative correlation with coronary heart disease. High density lipoproteins may be estimated by measuring cholesterol in the plasma fraction of d > 1.063 g/ml. A more practical approach is the specific precipitation of apolipoprotein B (apoB)-containing lipoproteins by sulfated polysaccharides and divalent cations, heparin-Mn2+ being the most commonly used combination. The present heparin-Mn2+ procedure was found to be reasonably specific and not often subject to large errors; however, 9% (primarily hypertriglyceridemic samples) of the 966 plasma samples treated with heparin-Mn2+ had obvious supernatant turbidity, indicating incomplete sedimentation of apoB-associated lipoproteins. Furthermore, 48% of the nonturbid supernates contained more than 1 mg/dl (mean 2.5 mg/dl) of apoB-associated cholesterol when measured by a radial immunodiffusion procedure, indicating slight overestimation of HDL cholesterol. Determination of the extent of the unprecipitated apoB-associated lipoproteins by sensitive radioimmunoassay and of the amount of precipitated high density lipoprotein by radial immunodiffusion assay of apolipoproteins A-I and A-II at various heparin and Mn2+ concentrations indicated that the usual heparin level (approximately 1.3 mg/ml) was adequate. However, a twofold increase in Mn2+ concentration to 0.092 M improved precipitation of the apoB-associated lipoproteins without excessive precipitation of high density lipoprotein from plasma. This increased Mn2+ level also provided improved sedimentation of the apoB-associated lipoproteins from hypertriglyceridemic plasma. Additional observations suggested that, for convenience, the heparin and Mn2+ can be added simultaneously as a combined reagent, that samples can be incubated for 10 minutes at room temperature before centrifugation, and that turbid supernates from hypertriglyceridemic samples can usually be made free of apoB-associated lipoproteins by centrifugation at 12,000 g for 10 minutes.
Supplementary key words coronary heart disease immunoassay lipoprotein quantitation
Submitted on February 22, 1977
Accepted on July 5, 1977

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A. Kondo, Y. Muranaka, I. Ohta, and T. Kanno
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S.-H. Bok, S.-H. Lee, Y.-B. Park, K.-H. Bae, K.-H. Son, T.-S. Jeong, and M.-S. Choi
Plasma and Hepatic Cholesterol and Hepatic Activities of 3-Hydroxy-3-methyl-glutaryl-CoA Reductase and Acyl CoA: Cholesterol Transferase Are Lower in Rats Fed Citrus Peel Extract or a Mixture of Citrus Bioflavonoids
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C. Marcoux, M. Tremblay, K. Nakajima, J. Davignon, and J. S. Cohn
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D. Heber, I. Yip, J. M Ashley, D. A Elashoff, R. M Elashoff, and V. L. W Go
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E. A. Stein, D. R. Illingworth, P. O. Kwiterovich Jr, C. A. Liacouras, M. A. Siimes, M. S. Jacobson, T. G. Brewster, P. Hopkins, M. Davidson, K. Graham, et al.
Efficacy and Safety of Lovastatin in Adolescent Males With Heterozygous Familial Hypercholesterolemia: A Randomized Controlled Trial
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L Rallidis, R P Naoumova, G R Thompson, and P Nihoyannopoulos
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D. F. Vine, J. C. L. Mamo, L. J. Beilin, T. A. Mori, and K. D. Croft
Dietary oxysterols are incorporated in plasma triglyceride- rich lipoproteins, increase their susceptibility to oxidation and increase aortic cholesterol concentration of rabbits
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M. S. Weintraub, I. Grosskopf, G. Charach, N. Eckstein, Y. Ringel, N. Maharshak, H. H. Rotmensch, and A. Rubinstein
Fluctuations of Lipid and Lipoprotein Levels in Hyperlipidemic Postmenopausal Women Receiving Hormone Replacement Therapy
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M. A. Ferguson, N. L. Alderson, S. G. Trost, D. A. Essig, J. R. Burke, and J. L. Durstine
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H. A. Whitley, S. M. Humphreys, I. T. Campbell, M. A. Keegan, T. D. Jayanetti, D. A. Sperry, D. P. MacLaren, T. Reilly, and K. N. Frayn
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J. L. Isaacsohn, M. Moser, E. A. Stein, K. Dudley, J. A. Davey, E. Liskov, and H. R. Black
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K. Sutton-Tyrrell, H. C. Lassila, E. Meilahn, C. Bunker, K. A. Matthews, and L. H. Kuller
Carotid Atherosclerosis in Premenopausal and Postmenopausal Women and Its Association With Risk Factors Measured After Menopause
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M. Miller, B. Teter, C. Dolinar, and A. Georgopoulos
An NCEP II Diet Reduces Postprandial Triacylglycerol in Normocholesterolemic Adults2
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F. Leyva, I. F. Godsland, M. Worthington, C. Walton, and J. C. Stevenson
Factors of the Metabolic Syndrome : Baseline Interrelationships in the First Follow-up Cohort of the HDDRISC Study (HDDRISC-1)
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S. F. Crouse, B. C. O'Brien, P. W. Grandjean, R. C. Lowe, J. J. Rohack, and J. S. Green
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T. A. B. Sanders, F. R. Oakley, G. J. Miller, K. A. Mitropoulos, D. Crook, and M. F. Oliver
Influence of n-6 versus n-3 Polyunsaturated Fatty Acids in Diets Low in Saturated Fatty Acids on Plasma Lipoproteins and Hemostatic Factors
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T. Dwyer, H. Iwane, K. Dean, Y. Odagiri, T. Shimomitsu, L. Blizzard, S. Srinivasan, T. Nicklas, W. Wattigney, M. Riley, et al.
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R Munro, E Morrison, A G McDonald, J A Hunter, R Madhok, and H A Capell
Effect of disease modifying agents on the lipid profiles of patients with rheumatoid arthritis
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N. Harris, V. Galpchian, J. Thomas, E. Iannotti, T. Law, and N. Rifai
Three generations of high-density lipoprotein cholesterol assays compared with ultracentrifugation/dextran sulfate–Mg2+ method
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H. C. Lassila, K. S. Tyrrell, K. A. Matthews, S. K. Wolfson, and L. H. Kuller
Prevalence and Determinants of Carotid Atherosclerosis in Healthy Postmenopausal Women
Stroke,
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S. F. Crouse, B. C. O'Brien, P. W. Grandjean, R. C. Lowe, J. J. Rohack, J. S. Green, and H. Tolson
Training intensity, blood lipids, and apolipoproteins in men with high cholesterol
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J. L. Isaacsohn, R. G. Bakker-Arkema, R. Fayyad, R. Whitcomb, and D. M. Black
Atorvastatin, a New HMG-CoA Reductase Inhibitor, Does Not Affect Glucocorticoid Hormones in Patients with Hypercholesterolemia
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[Abstract]
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S. T. Davidge, A. P. Signorella, C. A. Hubel, D. L. Lykins, and J. M. Roberts
Distinct Factors in Plasma of Preeclamptic Women Increase Endothelial Nitric Oxide or Prostacyclin
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M. Sugano and N. Makino
Changes in Plasma Lipoprotein Cholesterol Levels by Antisense Oligodeoxynucleotides against Cholesteryl Ester Transfer Protein in Cholesterol-fed Rabbits
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K. Handa, M. Takao, J. Nomoto, K. Oku, K. Shirai, K. Saku, and K. Arakawa
Evaluation of the Coagulation and Fibrinolytic Systems in Men with Intermittent Claudication
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V. Hachinski, C. Graffagnino, M. Beaudry, G. Bernier, C. Buck, A. Donner, J. D. Spence, G. Doig, and B. M. J. Wolfe
Lipids and Stroke: A Paradox Resolved
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D.R. Illingworth, E.A. Stein, R.H. Knopp, D.B. Hunninghake, M.H. Davidson, C.A. Dujovne, V.T. Miller, J.A. Tobert, P.M. Laskarzewski, J.L. Isaacsohn, et al.
A Randomized Multicenter Trial Comparing the Efficacy of Simvastatin and Fluvastatin
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R. R. Wing, R. W. Jeffery, and W. L. Hellerstedt
A Prospective Study of Effects of Weight Cycling on Cardiovascular Risk Factors
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E. Talbott, D. Guzick, A. Clerici, S. Berga, K. Detre, K. Weimer, and L. Kuller
Coronary Heart Disease Risk Factors in Women With Polycystic Ovary Syndrome
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E. T. Stevenson, K. P. Davy, and D. R. Seals
Hemostatic, Metabolic, and Androgenic Risk Factors for Coronary Heart Disease in Physically Active and Less Active Postmenopausal Women
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F. Tato, G. L. Vega, and S. M. Grundy
Bimodal Distribution of Cholesteryl Ester Transfer Protein Activities in Normotriglyceridemic Men With Low HDL Cholesterol Concentrations
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M. F. Muldoon, T. B. Herbert, S. M. Patterson, M. Kameneva, R. Raible, and S. B. Manuck
Effects of Acute Psychological Stress on Serum Lipid Levels, Hemoconcentration, and Blood Viscosity
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F. Tato, G. L. Vega, A. R. Tall, and S. M. Grundy
Relation Between Cholesterol Ester Transfer Protein Activities and Lipoprotein Cholesterol in Patients With Hypercholesterolemia and Combined Hyperlipidemia
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W. Insull Jr, D. Black, C. Dujovne, J. D. Hosking, D. Hunninghake, L. Keilson, R. Knopp, J. McKenney, E. Stein, A. J. Troendle, et al.
Efficacy and Safety of Once-Daily vs Twice-Daily Dosing With Fluvastatin, a Synthetic Reductase Inhibitor, in Primary Hypercholesterolemia
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K. A. Matthews, R. R. Wing, L. H. Kuller, E. N. Meilahn, and P. Plantinga
Influence of the Perimenopause on Cardiovascular Risk Factors and Symptoms of Middle-aged Healthy Women
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D. R. Illingworth, E. A. Stein, Y. B. Mitchel, C. A. Dujovne, P. H. Frost, R. H. Knopp, P. Tun, R. V. Zupkis, and R. A. Greguski
Comparative Effects of Lovastatin and Niacin in Primary Hypercholesterolemia: A Prospective Trial
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Pravastatin Multicenter Study Group II
Comparative Efficacy and Safety of Pravastatin and Cholestyramine Alone and Combined in Patients With Hypercholesterolemia
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D. B. Hunninghake, E. A. Stein, C. A. Dujovne, W. S. Harris, E. B. Feldman, V. T. Miller, J. A. Tobert, P. M. Laskarzewski, E. Quiter, J. Held, et al.
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R. R. Wing, K. A. Matthews, L. H. Kuller, D. Smith, D. Becker, P. L. Plantinga, and E. N. Meilahn
Environmental and Familial Contributions to Insulin Levels and Change in Insulin Levels in Middle-aged Women
JAMA,
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[Abstract]
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P. S. Bachorik, T. A. Cloey, C. A. Finney, D. R. Lowry, and D. M. Becker
Lipoprotein-Cholesterol Analysis during Screening: Accuracy and Reliability
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R. R. Wing, K. A. Matthews, L. H. Kuller, E. N. Meilahn, and P. L. Plantinga
Weight Gain at the Time of Menopause
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[Abstract]
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G. O. Perez, A. J. Mendez, R. B. Goldberg, R. Duncan, A. Palomo, E. DeMarchena, and S.L. Hsia
Correlates of Atherosclerosis in Coronary Arteries of Patients Undergoing Angiographic Evaluation
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Copyright © 1978 by the American Society for Biochemistry and Molecular Biology.
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