Di-tertiarybutylnitroxide (DTBN), which they have tried to use as a trapping agent to identify the species giving rise to the photo-induced EPR signals in photosynthetic materials, functions as a Hill reagent with spinach chloroplasts. Evidence is presented which indicates that the reduction of DTBN is affected by photosystem II of the electron transport system of spinach chloroplasts. The reduced form of DTBN, the hydroxylamine, undergoes a photo-oxidation with spinach chloroplasts. Possible explanations of this apparent inconsistency are presented. A product which could be ascribed to a chemical coupling reaction between the nitroxide and the radical species giving rise to the photo-induced EPR signals in spinach chloroplasts was not detected, even using radioactive tracer methods.