Luminol is one of the best known chemiluminescent cyclic hydrazides used in basic solution. Owing to the complexity of luminol oxidation, the mechanism of luminol chemiluminescence (CL), especially in aqueous solution, has not yet been fully elucidated. Recent theoretical computations have confirmed that luminol CL originates from the chemiexcitation of a 1,2-dioxane-3,6-dione dianion (CP2-). This seems to be inconsistent with the luminol oxidation in aqueous solutions, where only the decomposition of a monoanionic peroxyketal (L-OOH) is confirmed to yield CL. In this work, we theoretically investigated the complete decomposition of L-OOH and the pKa of key intermediates in aqueous solutions using (time-dependent) density functional theory. L-OOH first cyclizes to an endoperoxide monoanion (EP-). When pH < pKa (EP-), EP- directly decomposes by a retro-Diels-Alder (rDA) reaction to an aminophthalate monoanion (AP-) without CL activity. Moreover, when pH > pKa (EP-), EP- deprotonates to dianionic EP2-, which rapidly eliminates N2 to CP2-, inducing chemiexcitation. This conclusion is supported by the pKa (EP-) ≈ 8 estimated in this work, which is consistent with the pH-dependent profile of luminol CL observed in previous experiments. Thus, the pH-dependent CL determined by the acidity of endoperoxides may provide a theoretical basis to design new cyclic hydrazides with CL activity at different pH.