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Separation of Organic Acids in Blood by Partition Chromatography

Department of Dairy Science, University of Illinois, Urbana

ABSTRACT

The following chromatographic procedure, which involves the use of a silicic acid column, was developed primarily for the separation of beta-hydroxybutyric and other organic acids in the blood of ruminant animals. This procedure is basically similar to other silicic acid techniques (1, 3–7, 9), although its system of eluting solvents has been modified in an effort to improve the resolution of certain acids.

Silicic acid (Mallinckrodt 2847) is prepared for use by first removing the smaller particles, which markedly lessens the pressure required subsequently to produce the desired flow rate of solvent through the column. Such removal is achieved by suspending 250 g of the silicic acid in one liter of water. Let settle for 4 min, and discard the supernatant portion. Transfer the settled particles to a Büchner funnel, and suck dry. Complete the drying process in an oven at 110 C.

The eluting solvents are benzene, chloroform, and the following mixtures of tert-butanol in chloroform: 1, 2, 5, 8, 12, 16, 24, and 30% (v/v).