Heat treatment plays a decisive role in the microstructure of metallic materials. The effect of cooling rate changes caused by the quenching medium on the microstructure of steel materials should be clarified. In this study, the effect of the quenching cooling rate on the microstructure of two precipitation-hardened martensitic stainless steels was investigated. The mechanical properties and hydrogen embrittlement susceptibility effected by the changes in the microstructure were also analyzed. A slow tensile test and hydrogen pre-charging were carried out to obtain the hydrogen embrittlement susceptibility parameters of the specimens. The results show that the quenching cooling rate only affects specific microstructures, including the twin structure and misorientation angle. Before hydrogen charging, the mechanical properties of the precipitation-hardened martensitic stainless steels were not affected by changing the quenching cooling rate. After hydrogen charging, the hydrogen embrittlement susceptibility decreased as the quenching cooling rate increased.