DEAE-cellulose microcolumn chromatography coupled with automated assay: Application to the resolution of N-acetyl-β- d-hexosaminidase components

Abstract

A technique is described in which the entire effluent from a microcolumn is led directly into the flow-stream of an autoanalyzer and mixed with a fluorogenic substrate in buffer. The result is a continuous trace of the elution profile of enzyme activity on the recorder chart. Good separations of four N-acetyl-β- d-hexosaminidase components of human tissues were achieved in less than 30 min on 5 μl samples from 1% w v homogenates.