A Modified QuEChERS-HPLC Method for Detection of Polycyclic Aromatic Hydrocarbons in Zebrafish Embryos Exposed to Fine Particulate Matter

Maternal exposure to fine particulate matter (PM2.5), with an aerodynamic diameter of 2.5 µm or less, has been linked to spontaneous abortion and various congenital diseases in humans. However, the underlying mechanisms remain unclear. Zebrafish embryos are widely used to study the developmental toxicity of PM2.5 but measuring the internal levels of polycyclic aromatic hydrocarbons (PAHs), the primary toxic constituents of PM2.5, poses a challenge. In this study, we developed a Quick, Easy, Cheap, Effective, Rugged, and Safe High-performance Liquid Chromatography (QuEChERS-HPLC) method to assess the internal levels of the 16 PAHs listed in the US Environmental Protection Agency (EPA) priority pollutant list. Zebrafish embryos were treated with extractable organic matter (EOM) from PM2.5 for 2-72 h post fertilization (hpf). The QuEChERS method was employed for sample preparation, and HPLC-FLD/DAD (HPLC with fluorescence detector and diode array detector) was employed for the qualitative and quantitative analyses of PAHs. We successfully detected six PAHs (Phenanthrene, Anthracene, Fluorene, Acenaphthene, Fluoranthene, and Pyrene) in zebrafish embryos exposed to PM2.5. This method is crucial for investigating the mechanisms of PM2.5-induced developmental toxicity.