In this study, a chemiluminescence (CL) method for determination of glutathione (GSH) was developed with magnetic nanoparticle-decorated metal organic frameworks (Fe3O4 NPs@Cu-TATB). The composite material was synthesized via a hydrothermal method and glutathione (GSH) can be tested by both visual and chemiluminescence (CL) methods. The synthesized Fe3O4 NPs@Cu-TATB exhibited excellent catalytic activity in the luminol-H2O2 CL system. The mechanism revealed that three types of oxygen-containing radicals (ROS) was generated in this system. As GSH can reduce the catalytic effect of generated ROS radicals, the inhibiting CL signal was produced in the Fe3O4 NPs@Cu-TATB-luminol-H2O2 system. Based on the established CL system, the detection limits for GSH using CL and visual methods were found to be 0.3 µM and 0.7 µM, respectively. This low-cost and convenient detection method can be applied to the analysis of GSH content in human blood.