A RAPID PAPER CHROMATOGRAPHIC TECHNIQUE FOR SIMULTANEOUS ASSAY OF SEVERAL ENZYMES IN A TISSUE HOMOGENATE

In a program of clinical trials of enzyme-activated nitrogen mustard drugs in human cancer patients, it was desirable that the “activating enzymes” should be assayed in small biopsies of the tumors to aid in the choice of a suitable drug. Biochemical “test tube” methods of enzyme assay were sometimes time consuming and laborious and often required fairly large quantities of tissue, as when several enzymes had to be assayed in a tissue sample. In the present work, these difficulties were overcome by incubating substrates for esterase, amidase, and aminopeptidase with a single sample of tumor homogenate (containing 10 mg wet weight of tissue). The hydrolysis products were separated and estimated by a rapid method of paper chromatography and the total time involved in obtaining an assay of the three enzyme activities was not more than 3 hr.