Abstract
For the purpose to estimate total lipids and lipoprotein constituents in serum, an internal standard was introduced to electrophoresis on agarose gel. The internal standard was prepared by the procedure as shown in Fig. 1. Obtained dyed carbamylated human albumin (DC-Alb.) has similar color of lipid staining dye, Sudan black B and faster mobility than albumin as shown in Fig. 2. For electrophoresis, DC-Alb. solution was mixed with each serum sample with a definite ratio. The mixture was applied on agarose gel plate and electrophoresis, fixation and lipid staining with Sudan black B was performed as usual. Percent distribution of lipoprotein fractions and DC-Alb. was obtained by densitometry at 570nm. Percent density of DC-Alb. was correlated significantly with total lipids content determined with Fring's method (r=0.91). From regression equation obtained, total lipids can be calculated from densitometric scans of DC-Alb. and lipoproteins. The lipoprotein content estimated by multipling total lipids calculated as above and percent density of lipoprotein fractions on electrophoresis were correlated with those determined by analytical ultracentrifuge (r=0.84) as shown in Fig. 3. Thus, electrophorectogram with the internal standard can be used not only to estimate total lipids content but also to analyze lipoprotein constituents in the serum quantitatively, and is useful tool for investigation on lipoprotein metabolism in clinical practice for atherosclerosis or other lipid metabolism disorders.
