Journal of Lipid Research, Vol 27, 988-995, Copyright © 1986 by Lipid Research, Inc.
Sequential microultracentrifugation of lipoproteins in 100 microliters of serum
IR Kupke and S Worz-Zeugner
A method is described for sequential separation of high density, very low
density, and low density lipoproteins (HDL, VLDL, and LDL, respectively)
from 100 microliters of serum, using an air-driven ultracentrifuge
(Airfuge, Beckman). Cesium chloride was used for density adjustment.
Precision-within-series (coefficient or variation) depended on the
cholesterol concentration in the lipoprotein fractions, greater than 1
mmol/l, less than 2.3%. Recovery within-series was nearly 100%. The results
(mmol/l) correlate well with those from an electrophoretic-enzymatic
procedure (alpha-HDL: 1.49 +/- 0.34 vs. 1.48 +/- 0.33, r = 0.949;
pre-beta-VLDL: 0.58 +/- 0.42 vs. 0.59 +/- 0.45, r = 0.975; beta-LDL: 3.11
+/- 0.93 vs. 3.07 +/- 0.88, r = 0.990; n = 48). Recovery of lipoprotein
cholesterol from this group was 99.2 +/- 4.2%. A combination of
ultracentrifugation with high-performance thin-layer chromatography for
determination of lipoprotein-lipid profiles was achieved with recoveries of
98-101%, as evaluated from a group of healthy men (n = 31) and women (n =
38). The entire procedure is, therefore, suitable for compositional studies
on lipoproteins from small serum samples. In particular, capillary serum
from children of all ages, even from premature neonates, is quite adequate.
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