Lipidomic analysis has been proven to be a powerful technique to explore the underlying associations between xenobiotics and health status of organisms. Here, we established a strategy that combined the lipidomic analysis with high-throughput suspect contaminant screening technique with an aim to efficiently identify active xenobiotics in humans. Firstly, in the light of single liquid phase equilibrium of chloroform-methanol-water (15:14:2, v/v/v), we developed an efficient method that was able to simultaneously extract both polar and nonpolar lipids in serum samples. By use of this method, targeted and non-targeted lipid analyses were conducted for n=120 serum samples collected from Wuxi city, China. Secondly, we established a suspect database containing 1450 contaminants that have been previously reported in human samples, and contaminants in this database were screened in the same batch of serum samples by use of high-resolution mass spectrometry (HR-MS). Thirdly, the underlying associations between suspect contaminants and lipids were explored and discussed, and we observed that levels of some lipids were statistically correlated with concentrations of numerous contaminants. Among these active contaminants, 23 ones were identified on the basis of HR MS1 and MS2 characteristics, and these contaminants belonged to the classes of phthalates, phenols, parabens, or perfluorinated compounds (PFCs). Three active xenobiotics were fully validated by comparison with authentic standards, and they were perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS), and diethyl phthalate (DEP). There were statistically significant changes in levels of triglyceride (TG), lysophosphocholine (LPC), and sphingomyelin (SM) as peak areas of xenobiotics increase. We also observed that, among target lipid molecules, 18:0 lysophosphatidylethanolamine (LPE(18:0)) was very sensitive, and this lipid responded to exposure of various contaminants. Our present study provides novel knowledge on potential alteration of lipid metabolism in humans following exposure to xenobiotics, and provides an efficient strategy for efficiently identifying active xenobiotics in humans.