Determination of human serum lactate dehydrogenase isoenzymes by anion exchange chromatography

A pH-coupled salt gradient elution technique has been developed for the complete frationation of lactate dehydrogenase (LDH) isoenzymes in human sera by using a miniature polystyrene column packed with preequilibrated DEAE-cellulose to a bed of 8 cm x 0.7 cm. Five different buffers containing 0.02 M Tris (tris(hydroxymethyl)aminomethane hydrochloride) with increasing salt concentration and decreasing pH were used to separate the isoenzymes from 0.3 to 0.5 mL of sera absorbed on the column.