Background Existing methods for the detection of polycyclic aromatic hydrocarbons (PAHs) in blood have some shortcomings, such as complex and time-consuming operation, large consumption of organic solvents, and few detected kinds of PAHs, which need to be improved.
Objective This method ological study aims to establish a simple, effective, and environmentally friendly online solid phase extraction (SPE)-high performance liquid chromatography (HPLC) method for the determination of multiple PAHs in blood.
Methods The separation effects of methanol-water system and acetonitrile-water system on PAHs were compared, and the liquid chromatography conditions were further optimized. The affinities of Hypersep Hypercarb column, Dionex InGuard HRP column, Hypersep Retain PEP column, and C18 online SPE column for PAHs were compared to select SPE column and determine valve switching procedure. In addition, the induction effects of different inducers (dichloromethane, trichloromethane, and and toluene) were compared, and the effects of different dosages of acetonitrile, dichloromethane, and inorganic sodium chloride on induced phase change extraction efficiency were investigated by four-factor and three-level orthogonal test. Method ological validation was performed by applying the established method to the determination of PAHs in blood samples of eight mechanical workers and six general people.
Results The C18 online SPE column showed good separation of PAHs when being eluted with methanol-water system. The results of orthogonal test showed that 0.50 mL acetonitrile, 0.05 mL dichloromethane, and no sodium chloride were the best pretreatment conditions. The linear relationships of 15 PAHs were good in the range of 1.00~50.00 μg/L, the determination coefficients were above 0.999, and the detection limits (signal-to-noise ratio, S/N=3) were 0.01-0.20 μg/L. The average recovery rates were 90.8%-107.0%, and the relative standard deviations (n=6) were 1.6%-4.4%. The blood samples of eight mechanical workers and six general people were determined by this method, detection rates of low-ring PAHs were high, and the detection rates of high-ring PAHs were low; three more high-ring PAHs were detected in the mechanical workers than in the general subjects.
Conclusion The proposed method can simultaneously determine the contents of 15 PAHs in serum, and is simple, efficient, accurate, reliable, and environmentally friendly. It can provide method ological support for exposure risk assessment and toxicology research of PAHs.
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