The aims of the present study were to develop environmental and low cost methods for detection of tetracyclines (TCs) concentrations in the fish by high performance liquid chromatography with chemiluminescence.Luminol chemiluminescence (CL) could be generated by generated active oxygen species of photo-induced nano-titanium dioxide (TiO₂) instead of the traditional chemiluminescent oxidation reagents.Metal ions could catalyze this reaction.Based on the complexation between Fe²⁺ and TCs that remarkably decreased the Fe²⁺-catalyted luminol CL,the determination of TCs was proposed.The results indicated that the limit of quantitation for tetracycline,chloroteracycline and doxycycline were 10.0,30.0 and 10.0 mg/kg,respectively.The proposed method could be applied to the determination of the TCs in the fish,and meet the requirements of residue detection.Nano TiO₂ was used instead of traditional chemiluminescent oxidation reagents to avoid the application of some toxic reagents and chemical reagent pollution,and had the advantages of environmental protection and low price.