Cathodic electrochemiluminescence (ECL) of a luminol (or its analogues)-dissolved oxygen (O 2 ) system is an ideal alternative to ECL of the traditional luminol-hydrogen peroxide (H 2 O 2 ) system, which can efficiently avoid the self-decomposition of H 2 O 2 at room temperature. However, the mechanism for the generation of cathodic ECL by the luminol (or its analogues)-O 2 system is still ambiguous. Herein, we report the study of cathodic ECL generation by the L012-O 2 system at a glassy carbon electrode (GCE). The types of reactive oxygen species (ROS) involved generated during ECL reactions were verified. A possible reaction mechanism for the system was proposed and the rate constants of related reactions were estimated. Furthermore, several intermediates of L012 involved in the proposed pathways were validated by electrochemistry-coupled mass spectrometry. Finally, the cathodic ECL system was successfully used for measuring the antioxidant capacity of commercial juice with Trolox as a standard.