Nitration can enhance the allergenicity of proteins. The nitration status of house dust mite (HDM) allergens in indoor dusts, however, remains to be elucidated. In the study, site-specific tyrosine nitration degrees of the two important HDM allergens Der f 1 and Der p 1 in indoor dust samples were investigated by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The measured concentrations of native and nitrated allergens in the dusts were in the range of 0.86–29 μg g⁻¹ for Der f 1 and from below the detection limit to 29 μg g⁻¹ for Der p 1. Site-specific analysis revealed that all ten tyrosine residues in Der f 1 and Der p 1 were nitrated to different degrees in the investigated samples. The preferred nitration sites were Y₅₆ in Der f 1 and Y₃₇ in Der p 1 with the nitration degrees of 7.6–84% and 17–96% among the detected tyrosine residues, respectively. The measurements reveal high site-specific nitration degrees for tyrosine in Der f 1 and Der p 1 detected in the indoor dust samples. Further investigations are required to find out if the nitration really aggravates the health effects of HDM allergens and if the effects are tyrosine site-dependent.