Unmanned Aerial Systems (UAS) Photogrammetry has become widely used for spatial data acquisition. Nowadays, RTK (Real Time Kinematic) and PPK (Post Processed Kinematic) are the main correction methods for accurate positioning used for direct measurements of camera station coordinates in UAS imagery. Thus, 3D camera coordinates are commonly used as additional observations in Bundle Block Adjustment to perform Global Navigation Satellite System-Assisted Aerial Triangulation (GNSS-AAT). This process requires accurate Interior Orientation Parameters to ensure the quality of photogrammetric intersection. Therefore, this study investigates the influence of on-site camera calibration with a sub-block of images on the accuracy of spatial data obtained by PPK-based UAS Photogrammetry. For this purpose, experiments of on-the-job camera self-calibration in the Metashape software with the SfM approach were performed. Afterward, experiments of GNSS-Assisted Aerial Triangulation with on-site calibration in the Erdas Imagine software were performed. The outcomes show that only the experiment of GNSS-AAT with three Ground Control Points yielded horizontal and vertical accuracies close to nominal precisions of the camera station positions by GNSS-PPK measurements adopted in this study, showing horizontal RMSE (Root-Mean Square Error) of 0.222 m and vertical RMSE of 0.154 m. Furthermore, the on-site camera calibration with a sub-block of images significantly improved the vertical accuracy of the spatial information extraction.