Abstract
To assess the limits of the heparin-MnCl2 precipitation method for quantitation of α-lipoprotein cholesterol (C-HDL), effects of varying final MnCl2 and heparin concentrations were studied, and the precipitation method was compared to preparative ultracentrifugation. In 65 parallel plasma aliquots, C-HDL\(\left( {\bar X \pm SE} \right)\) determined by ultracentrifugation (54.3±1.8 mg/dl) correlated significantly (r=0.98, P<0.001) with the precipitation method (56.0±1.9 mg/dl). C-HDL by ultracentrifugal and precipitation methods were also similar in 16 subjects with triglycerides ranging from 150 to 312 mg/dl (41.4±2.6, 43.4±2.8, r=0.97, P<.001). A constant amount of cholesterol in the supernatant was measured over a final MnCl2 range of 0.046–0.23M, and cholesterol values in the supernatant at final MnCl2 concentrations of 0.046, 0.05, and 0.055M did not differ from each other, P>0.1. However, cholesterol levels in the supernatant at final MnCl2 concentration of 0.042M differed from those at concentrations of 0.046, 0.05, and 0.055M, P<0.05, and the amount of supernatant cholesterol increased as the final MnCl2 concentration was reduced from 0.042 to 0.02M. A constant amount of cholesterol in the supernatant was measured over a heparin concentration range of 92–734 USP units/ml. The final MnCl2 and heparin concentrations of 0.046M and 184 USP units/ml, which are incorporated in widely used procedures, gave C-HDL values for the precipitation method which were in close agreement with the ultracentrifugal method. There is no evidence for a heparin-Mn++ precipitation of HDL and systematic underestimation of HDL by the precipitation method. However, the final MnCl2 concentration is very near the minimum required for accurate measurement of C-HDL. To preclude incomplete precipitation of low and very low density lipoproteins by insufficient manganese concentration, an increase of the manganese concentration should be considered.
Similar content being viewed by others
References
Nikkila, E., Scand. J. Clin. Lab. Invest., Suppl. 5:1–101 (1953).
Gofman, J.W., H.B. Jones, F.T. Lindgren, T.P. Lyon, H.A. Elliot, and B. Strisower, Circulation 2:161 (1950).
Barr, D.P., E.M. Russ, and H.A. Eder, Am. J. Med. 11:480 (1951).
Jencks, W.P., M.R. Hyatt, M.R. Jetton, T.W. Mattingly, and E.L. Durrum, J. Clin. Invest. 35:980 (1956).
Miller, G.J., and N.E. Miller, Lancet 1 (7897): 16 (1975).
Castelli, W.P., J.T. Doyle, T. Gordon, C. Hames, S.B. Hulley, A. Kagan, D. McGee, W.J. Vicic, and W.J. Zukel, Circulation 51,52:Supplement II, II-96 (1975).
Glueck, C.J., R.W. Fallat, F. Millett, P. Gartside, R.C. Elston, and R.C.P. Go, Metabolism 24:1243 (1975).
Glueck, C.J., R.W. Fallat, and M. Spadafora, Circulation 51,52:supplement II, II-272 (1975).
Burstein, M., and J. Samaille, Clin. Chim. Acta 5:609 (1960).
Fredrickson, D.S., R.I. Levy, and F.T. Lindgren, J. Clin. Invest. 47:2446 (1968).
Manual of Laboratory Operations, Lipid Research Clinics Program, Vol. 1, Lipid and Lipoprotein Analysis, HEW Publication (NIH) 75-628 (1974).
Friedewald, W.T., R.I. Levy, and D.S. Fredrickson, Clin. Chem. 18:499 (1972).
Srinivasan, S.R., B. Radhakrishnamurthy, and G.S. Berenson, Arch. Biochem. Biophys. 170:334 (1975).
Lutmer, R.F., D. Parsons, C.J. Glueck, J.A. Morrison, L. Stewart, J.B. Brazier, C.R. Buncher, and T.T. Ishikawa, J. Lipid Res. 15:611 (1974).
Hatch, F.T., and R.S. Lees in “Advances in Lipids Research,” Edited by R. Paoletti and D. Kritchevsky, Academic Press, New York, NY, 1968, p.35.
Ishikawa, T.T., J. MacGee, J.A. Morrison, and C.J. Glueck, J. Lipid Res. 15:286 (1974).
Burstein, M., and H.R. Scholnick, in “Advances in Lipid Research,” Edited by R. Paoletti, and D. Kritchevsky, Academic Press, New York, NY, 1973, pp. 67–108.
Wilson, D.E., and M.J. Spiger, J. Lab. Clin. Med. 82:473 (1973).
Flaschka, H., and A.M. Amin, Mikrochim. Acta 4141:420 (1953).
Nader, H.B., N.H. McDuffie, and C.P. Dietrich, Biochem. Biophys. Res. Commun. 57:2 (1974).
Noble, R.P., F.T. Hatch, J.A. Mazrimas, F.T. Lindgren, L.C. Jenson, and G.L. Adamson, Lipids 4:55 (1969).
Overall, J.E., and D.K. Spiegel, Psychol. Bull. 72:311 (1960).
Rider, A.K., R.I. Levy, and D.S. Fredrickson, Circulation 42:10 (1970).
Ehnholm, C., H. Garoff, K. Simons, and H. Aro, Biochim. Biophys. Acta 236:431 (1971).
Author information
Authors and Affiliations
Additional information
A portion of this work was supported by General Clinical Research Center Grant No. RR00068-14, and a portion of it was done during Dr. Glueck’s tenure as an Established Investigator of the American Heart Association, 1971–1976.
About this article
Cite this article
Ishikawa, T.T., Brazier, J.B., Steiner, P.M. et al. A study of the heparin-manganese chloride method for determination of plasma α-lipoprotein cholesterol concentration. Lipids 11, 628–633 (1976). https://doi.org/10.1007/BF02532877
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02532877