A novel targeted high-throughput workflow for serum chemical mixture profiling in population exposome-wide association studies.

Simultaneous determination of environmental pollutants with complex compositions and wide-ranging acid dissociation constants in biological matrices presents a significant analytical challenge. This study developed a highly sensitive and broad-spectrum analytical method based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). By employing phospholipid removal purification (PRP) technology and optimizing mobile phase pH conditions, we achieved simultaneous quantification of 230 environmental pollutants in serum. Comparative evaluation of sample pretreatment techniques revealed that low-temperature concentration caused preservative loss, while liquid-liquid extraction showed poor recovery for multiple compound classes. The PRP method was selected as the optimal approach due to its superior performance in reducing matrix effects compared to direct protein precipitation. Method validation demonstrated accurate quantification of all target analytes (relative standard deviation, RSD <15.1%), with 98.3% of compounds exhibiting satisfactory recoveries (80-120%). Analysis of serum samples from the Fuqing cohort participants detected 22 contaminants with detection frequencies >30%, among which PFHpA and PFOA were identified as risk factors for hyperuricemia. Notably, we provide the first evidence of a dose-response relationship between dehydroacetic acid exposure and hyperuricemia prevalence in a human population. This study established a robust solution for simultaneous screening of exposome-scale pollutants in complex biological samples.