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The aim of this study was to examine the consistency of ultra performance liquid chromatography-tandem mass spectrometry (UPLC-TMS) in detecting the levels of para-arachidonic acids (PAAs) among differently processed plasma/serum samples. Ethylenediaminetetraacetic acid (EDTA)-K2, sodium citrate, heparin lithium, coagulant/separation gel, and coagulant-free vacuum blood-sampling tubes were used to collect the fasting blood samples from 15 volunteers. All blood samples were subjected to solid-phase extraction using an Oasis HLB mu Elution 96-well plate, and UPLC-TMS was used to detect 19 types of PAAs in the blood samples. Within the plasma samples, the concentrations of 5, 6-DHET; 11, 12-epoxyeicosatrienoic acid (EET); 5-hydroxyeicosatetraenoic acid (HETE); leukotriene B4 (LTB4); plasma thromboxane B2 (TXB2); and 12-HETE were significantly higher in the heparin lithium group than in the EDTA-K2 and sodium citrate groups. Within the serum samples, the concentration of LTB4 was significantly higher in the coagulant/separation gel group than in the coagulant-free group, while that of TXB2 was significantly higher in the coagulant-free group than in the coagulant/separation gel group. The levels of some types of PAAs in differently processed plasma/serum samples were inconsistent, and the concentrations of 5, 6-DHET; 5-HETE; 12-HETE; TXB2; and LTB4 were significantly higher in the two serum samples and the heparin lithium group than in the EDTA-K2 and sodium citrate groups.