Photometric measurement of lipoprotein-cholesterol after agarose electrophoresis: comparison with single-spin ultracentrifugal analysis.

ABSTRACT

A method is presented for the evaluation of lipoprotein-cholesterol after fractionation of 20 microliter of serum in agarose electrophoresis in a discontinuous buffer system. The prestained lipoprotein bands are separated in less tha 1 h, cut out and extracted with butanol. The cholesterol in the extract is measured by a simple colorimetric method. The recoveries of lipoprotein cholesterol were 95.3% +/- 13.0 (S.D.), and the average coefficient of variation was 8% for alphalipoprotein, 6.4% for betalipoprotein and 8.0% for pre-betalipoproteins. Lipoprotein cholesterol concentrations, determined by the proposed method correlated well with those analyzed following ultracentrifugation. The method can handle up to 4 serum samples run in duplicate per day and only requires equipment available in conventional clinical laboratories, and stable reagents.