Luminescence spectra of aqueous solutions containing a fixed concentration of tetracycline (TC) and increasing concentrations of Eu3+ were recorded both in the absence and presence of hydrogen peroxide (H2O2). In H2O2-free solutions in which the Eu/TC molar ratio was varied from 1 : 1 to 8 : 1, the 5D0 --> 7F0 transition consisted of only one peak at 580 nm. In the presence of H2O2, an extra peak appeared in the spectrum at 578 nm when the Eu/TC molar ratios were above 2.5. A detailed analysis of this spectral region revealed that at lower Eu/TC molar ratios (up to 2 : 1), the 5D0 --> 7F0 transition experienced a slight blue shift. This indicates that at low Eu/TC molar ratios, the presence of H2O2 leads to two different environments of the trivalent europium ions, which most likely form bridged peroxide complexes with hydrogen peroxide (mu-H2O2 ligand). Luminescence spectra measured in the presence of molybdate ions, which catalytically decompose H2O2, led to the disappearance of the extra europium(III) site that was formed in the presence of H2O2. The intensity of the hypersensitive 5D0 --> 7F2 transition did not linearly depend on the H2O2/TC molar ratio. For H2O2/TC ratios up to 10, a sharp linear increase of the peak intensity was observed, but with further increase of the H2O2 concentration, the intensity remained nearly constant. For H2O2/TC ratios above 100, the intensity of this transition even started to decrease, which limits the use of the (tetracycline)europium(III) system to quantify hydrogen peroxide in solution.