Glutathione sulfhydryl oxidase and catalase are separately immobilized onto oxirane-acrylic beads (Eupergit-C). Both types of beads are double-layered into a small polymer column. The flow-injection analysis system for glutathione is assembled with the column and a calorimetric device for enzymatically generated heat. The calibration graph for glutathione is linear from 0.5 to 10 mM for 0.2 ml samples. The assay takes less than 6 min. The relative standard deviation for 5.0 mM glutathione is 1.2% ( n = 5).