In competition-based experiments, 1,4-benzoquinone (BQ) has been a frequently applied compound for determination of the role of O2− radicals. In those studies, only the influence of BQ (as a competitive O2− scavenger) on the degradation of a model compound (substrate) was investigated. Our goal was to develop a new, simple method monitoring a characteristic product of the reaction between the superoxide radical ion and BQ during heterogeneous photocatalysis. Interestingly, based on our results, if the concentration of BQ exceeds that of dissolved oxygen, it reacts with photogenerated electrons rather than with O2− radicals. During this process, it is quantitatively reduced to 1,4-hydroquinone (H2Q), the degradation of which was found to be dependent of the presence of O2− radicals. The photocatalytic transformation of both compounds (BQ, H2Q) was systematically investigated in anoxic and oxic atmosphere. Two new methods requiring only fluorescence measurements have been suggested: one for the determination of electrons with BQ (under anaerobic conditions) and another for the measurement of O2− radicals with H2Q.