In this paper, the μGC-IL/MOF and the μGC-IL were prepared using [P66614][NTf2]/ZIF-8 and [P66614][NTf2] as the stationary phase, respectively. [P66614][NTf2]/ZIF-8 composite stationary phase material has high specific surface area and porous structure, which increases the diffusion of gas molecules in the column. Compared to μGC-IL, μGC-IL/MOF can separate n-alkanes mixture (C5–C12) and lung cancer biomarkers (pentane, isoprene, acetone, benzene, 2-pentanone) with higher resolution, and the resolution (R) of pentane and isoprene was increased by 257.00% in particular. The μGC-IL/MOF can separate lung cancer biomarkers in about 5 min with optimal carrier gas velocity and column temperature. The retention times of pentane, isoprene, acetone, benzene, and 2-pentanone were 0.884 min, 1.246 min, 1.634 min, 2.204 min, and 3.049 min, respectively. The resolutions of adjacent peaks were 1.785, 1.525, 2.521, and 3.514, respectively. Which meets the requirements of quantitative analysis (R > 1.5). Therefore, the μGC-IL/MOF is expected to be integrated into portable devices for environmental monitoring and medical diagnosis in the future.