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Polydopamine nanospheres (PDA) were designed to serve as a new substrate for surface-enhanced desorption/ionization mass spectrometry (SELDI-MS). Compared with conventional organic matrices, the PDA substrate showed superior LDI performance for analyzing a wide variety of environmental pollutants, including polycyclic aromatic hydrocarbons, bisphenols, benzophenones, sulfonamides, perfluorinated compounds and estrogens. Benzoapyrene was used to evaluate the ability of quantitative analysis and its corresponding limit of detection (LOD) of as low as 0.1 ng was achieved. High sensitivity and good reproducibility of PDA-based SELDI-MS allowed us to determine ultratrace PAHs in airborne particulate matters (PM2.5), and the corresponding concentration of BaP in different PM2.5 were 5.32, 8.97 and 9.79 ng m-3. Significantly, PDA exhibits the characteristics of simple synthesis, low cost, non-toxicity and less matrix interference, which provides the possibility for the sensitive analysis of organic small molecule pollutants at low concentrations in complex environmental samples.