The vertical profile of atmospheric temperature is crucial for understanding energy transfer within the climate system. The Moderate Resolution Imaging Spectroradiometer (MODIS) atmospheric profile product (MOD07_L2) provides vertical temperature profiles at 5 km spatial resolution. However, comprehensive validation of its absolute accuracy—particularly for lower tropospheric stability (LTS) and temperature inversion layer (TIL) detection—remains limited. Therefore, this study conducted systematic evaluations using sounding data from 74 stations across mainland China spanning 2003–2020, encompassing both daytime and nighttime periods. Results indicated that absolute retrieval accuracy was generally superior at nighttime compared to daytime, with MOD07_L2 demonstrating better performance in upper layers (400–620 hPa) than in lower layers (850–1000 hPa). Nonetheless, retrieval errors at the 700 hPa level exhibited a more pronounced bias (B) relative to other pressure levels. In contrast, LTS accuracy was higher during daytime (r = 0.74, B = 0.04 °C, RMSE = 4.52 °C) than at nighttime (r = 0.57, B = 0.3 °C, RMSE = 6.78 °C).For TIL, MOD07_L2 displayed limited detection capability, achieving an overall detection rate of 6.6 % during daytime and 13.1 % at nighttime. This study provides essential data support for atmospheric science, hydrology, and ecology fields that rely on high-resolution vertical temperature profiles.