Abstract: Deciphering hydrocarbon generation and accumulation stage is of significance to understand oil and gas evolution and seek exploration targets. Taking the Upper Paleozoic buried-hills in the Huanghua Depression, Bohai Bay Basin, as a case study, hydrocarbon generation environment and detailed accumulation process are revealed by fluid inclusions observations, Laser Raman spectroscopy, Fourier Infrared spectroscopy, and K–Ar isotope measurements. The results show that both oil and gas inclusion were captured in the quartz overgrowth, dissolved feldspar and calcite microfractures, showing blue to dark brown fluoresce. The grains containing oil inclusions index (GOI) of oil, oil & gas and gas being 25%, 65%, and 10% and the inclusions with abundant methyl groups and short chains, both indicate high thermal maturity. One series of fluids inclusion is generally observed, evidenced by the concentrated homogenization temperature of 135–145 °C and salinity of 3%–15 w.t.% NaCl equiv, indicating one primary charging stage. The gas and gas & liquid inclusions mainly contain CH4, with also peaks indicating CO2 and N2. The Carboniferous and Permian biomarkers show reducing environment with brackish water, with organic matter sources both from marine and continental. The relative content of ααα20RC27, ααα20RC28, and ααα20RC29 exhibit source contributions both from algae and higher plants, and mainly of II2 to III kerogen. Both coal derived gas and oil associated hydrocarbons are identified from most of the buried-hills. Combining the fluid homogenization temperature and salinity, as well as the thermal evolution history, the hydrocarbon generated from the Upper Paleozoic was concentrated at the end of the Eocene (40 Ma±), while the beginning of charging is 60 Ma±. The Wumaying Buried-hill is of only coal derived gas and has potential for inner coal measure natural gas exploration. The results provide a detailed understanding of hydrocarbon accumulations in the study area, which can also be reference for improving petroleum exploration efficiency in similar basins.