Titanium dioxide (TiO2) nanoparticles (NPs) have been widely used in industrial and consumer products. Comprehensive and accurate detection, characterization, and quantification of TiO2 NPs are important for understanding the specific property, behavior, fate, and potential risk of TiO2 NPs in natural and engineered environments. This review provides a summary of recent analytical studies of TiO2 NPs and their aggregation, coagulation, flocculation, sedimentation, stabilization under a wide range of conditions and processes. Much attention is paid on sample preparation prior to an analytical procedure, analysis of particle size, morphology, structure, state, chemical composition, surface properties, etc., via measurements of light scattering and zeta potential, microscopy, spectroscopy, and related techniques. Recently, some advanced techniques have also been explored to characterize TiO2 NPs and their behaviors in the environment. Many issues must be considered including distinction between engineered TiO2 NPs and their naturally occurring counterparts, lack of reference materials, interlaboratory comparison, when analyzing low concentrations of TiO2 NPs and their behaviors in complex matrices. No “ideal” technique has emerged as each technique has its own merits, biases, and limitations. Multi-method approach is highlighted to provide in-depth information. Improvements of analytical method for determination of TiO2 NPs have been recommended to be together with exposure modelers and ecotoxicologists for maximum individual and mutual benefit. Future work should focus on developing analytical technology with the advantages of being reliable, sensitive, selective, reproducible, and capable of in situ detection in complicated sample system. Display omitted •Essential to assess behavior of TiO2 NPs in natural and engineered environment.•Challenging to determine TiO2 NPs at low concentrations in complex matrices.•Efforts to characterize TiO2 NPs in aggregate, coagulate, flocculate, sediment.•Important to stabilize TiO2 NP to control particle fate, effect, and application.•More advanced methods in need to investigate TiO2 NPs in complex matrices.