Objective To investigate the concentration of perfluoroalkyl and polyfluoroalkyl substances (PFASs) in the serum of the general population in Wuhan, and to discover the influencing factors of the concentrations of PFASs in serum. Methods Total of 67 people who had physical examination in a certain hospital in Wuhan was selected as the study object. The concentration of 12 kinds of PFASs in peripheral serum were determined by ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-TQ/MS). The correlation between general demographic characteristics and PFASs concentration in serum was discovered by multiple linear regression analysis. Results The average concentration of the 12 kinds of PFASs in the serum of the general population in Wuhan was 11.60 ng/mL, which was lower than those in previous studies. Perfluorooctane sulphonate(PFOS) and perfluoro octanoic acid(PFOA) were the main contributors, accounting for 45.03% and 31.86% respectively. The concentration of main compounds including PFOS, PFOA, perfluorohexanesulfonic acid(PFHxS), and perfluorononanoic acid(PFNA) in the serum of male population was significantly higher than that of female population, with the β value and 95% confidence interval (95%Cl) of 0.708 (0.313,1.104), 0.518(0.069,0.967), 0.724(0.388,1.059) and 0.684(0.399,0.968) respectively. At the same time, PFOS, PFHxS and PFNA in serum were positively correlated with age, with the β value and 95%Cl confidence interval of 0.062(0.042,0.081), 0.035(0.019,0.052) and 0.030(0.016,0.045) respectively. Conclusion Gender and age have influence on PFASs concentration in serum, and these factors should be taken into account for human exposure assessment of PFASs.