In conclusion, the present study established a robust LC-MS/MS method for simultaneously detecting 8,15-DiHETE, HDA, DHK-PGA2, RCL, OA, and 16OHHA, and it is the first to report that serum 8,15-DiHETE, HDA, DHK-PGA2 and OA were elevated in T2DM(+) HCC, and the biomarker signature based on HDA, DHK-PGA2 and AFP shows great diagnostic efficacy for T2DM(+) HCC. Here, AFP is linked to type 2 diabetes mellitus.