Journal of Lipid Research, Vol. 12, 420-433, July 1971
Copyright © 1971 by Lipid Research, Inc.

Quantitation of serum lipoproteins by electrophoresis on agarose gel: standardization in lipoprotein concentration units (mg/100 ml) by comparison with analytical ultracentrifugation

S. B. Hulley , S. G. Cook , W. S. Wilson , M. Z. Nichaman , F. T. Hatch , and F. T. Lindgren

Community Medicine Program and the Metabolic Unit, U.S. Public Health Service Hospital, San Francisco, California 94118; Biomedical Division, Lawrence Radiation Laboratory, University of California, Livermore, California 94550; and Donner Laboratory, University of California, Berkeley, California 94720

Lipoprotein electrophoresis on agarose gel has been modified to allow estimation of the absolute quantity of each fraction. The reproducibility of the method is illustrated by 12 determinations in a single day on serum from one normal subject: mean total dye uptake was 302 ± 9 (sd "corrected dye units," and the percentages of ßbeta;-, pre-ßbeta;, and -lipoprotein were 56.1 ± 0.9, 29.1 ± 0.4, and 14.8 ± 0.7, respectively. Reproducibility over a period of 8 months was also demonstrated.

Serum lipoproteins of five normal and 15 hyperlipidemic individuals determined by this technique were compared with values obtained by analytical ultracentrifugation. The correlation coefficients were: 0.993 for pre-ßbeta;-LP vs. VLDL, 0.978 for ßbeta;-LP vs. LDL, and 0.867 for -LP vs. HDL. Lipoprotein values obtained by paper electrophoresis were also correlated with those of the analytical ultracentrifuge, but to a lesser degree (r = 0.956, 0.691, and 0.786, respectively). Values for LDL and VLDL which were measured by refractometry after preparative ultracentrifugation were very similar to those obtained from the analytical ultracentrifuge. Serum triglyceride concentration was highly correlated (r = 0.972) with the agarose values for pre-ßbeta;-LP; serum cholesterol concentration was correlated (r = 0.673) with ßbeta;-LP. It is proposed that the standard curves of the comparisons with the analytical ultracentrifugal values be used to convert the corrected dye units of electrophoresis on agarose gel to mg/100 ml of specific lipoprotein.

Supplementary key words cholesterol • gravimetric total lipids • paper electrophoresis • refractometry • triglyceride

Submitted on October 26, 1970
Accepted on February 18, 1971

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				H. Hidaka, M. Tozuka, B. Meyer, K. Yamauchi, M. Sugano, T. Nakabayashi, and T. Katsuyama Characterization of Triglyceride Rich Lipoproteins with Very Light Density by Ultracentrifugation and Agarose Gel Electrophoresis using Triglyceride- and Cholesterol-Staining Ann. Clin. Lab. Sci., April 1, 2003; 33(2): 167 - 178. [Abstract] [Full Text] [PDF]