The finite element modes of the quartz tuning fork in the Quartz Enhanced Photoacoustic Spectroscopy (QEPAS) system were calculated and the first 6 modes and modal frequencies of the quartz tuning fork were obtained. The 4th order symmetrically oscillating vibration mode was recognized as an effective vibration. Single factor method was used to analyze the effects of fork’s arm length l1, fork’s arm width w1, fork’s arm thickness t, fork’s arm cutting angle θ, fork’s arm diameter d and fork’s arm round hole height h on low effective resonance frequency (Fre). The results of the sensitivity sensitivity were as follows: l1>w1>d>θ>t>h. Considering the actual design situation, four design parameters of quartz tuning fork l1, w1, d and h were screened. Using Box-Behnken experimental design and RSM (Response Surface Methodology) method, Fre was set as a function target to establish l1, w1, d and h quadratic regression response surface model, and get the interaction between the parameters. The Design-Expert software was used to inverse the design parameters of the response surface model. The results showed that the error in the calculation area of 15 000 Hz≤Fre≤25 000 Hz is small, and basically meets the calculation requirements of QEPAS system. The proposed research and design methods have some generality, which can provide references for the design of quartz tuning fork structure in QEPAS system.