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Journal of Lipid Research, Vol. 44, 2193-2201, November 2003
Copyright © 2003 by American Society for Biochemistry and Molecular Biology

Methods

A novel and simple method for quantification of small, dense LDL

Tsutomu Hirano^1,*, Yasuki Ito, Haruhisa Saegusa and Gen Yoshino

^* First Department of Internal Medicine, Showa University School of Medicine, Tokyo, Japan
Research and Development Department, Denka Seiken Co., Ltd., Tokyo, Japan
Department of Laboratory Medicine, Toho University School of Medicine, Tokyo, Japan

¹ To whom correspondence should be addressed. e-mail: hirano{at}med.showa-u.ac.jp

A preponderance of small, dense (sd) LDL is strongly associated with the development of coronary heart disease, but the method for the measurement of sd LDL is too laborious for clinical use. We report a simple method for the quantification of sd LDL that is applicable to an autoanalyzer. This method consists of two steps: first, to precipitate the lipoprotein of density (d) <1.044 g/ml using heparin-magnesium; and second, to measure LDL-cholesterol in the supernatant by the homogenous method or apolipoprotein B (apoB) by an immunoturbidometric assay. The cholesterol and apoB values obtained by the precipitation method (45 ± 26 and 33 ± 20 mg/dl, respectively) were similar to those obtained in the lipoprotein (d = 1.044–1.063) separated by ultracentrifugation (42 ± 22 and 31 ± 17 mg/dl, respectively), and there was an excellent correlation between the two methods for sd LDL-cholesterol (y = 1.05X + 1, r = 0.88, n = 69) and apoB (y = 1.07X, r = 0.90). Sd LDL values had a significant inverse correlation with LDL size. A high correlation was found between sd LDL-cholesterol and apoB values (r = 0.94). Sd LDL value was related to triglyceride, apoB, and LDL-cholesterol, but not to the buoyant LDL level.

These results suggest that this precipitation method is a simple and rapid method for the measurement of sd LDL concentration.

Supplementary key words cholesterol • apolipoprotein B • precipitation

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