This study aims to qualitatively identify saponin components in Panacis Majoris Rhizoma and establish a method for the quantitative determination of multiple saponin constituents. UPLC-Q-TOF-MS/MS technology was employed to scan Panacis Majoris Rhizoma in both positive and negative ion modes. The UNIFI natural product information platform was used for the qualitative identification and analysis of chemical components present in Panacis Majoris Rhizoma. A gradient elution method was applied using a mobile phase consisting of 0.1% phosphoric acid in water and acetonitrile. The column temperature was set at 30 °C with a flow rate of 0.3 mL/min for the quantitative determination of multiple saponin constituents. A total of 39 chemical components were identified in Panacis Majoris Rhizoma, including 37 saponin compounds and two saponin backbone. The fragmentation patterns of saponins were summarized, and a UPLC method was established for the simultaneous determination of ginsenoside Rb₁, ginsenoside Ro, ginsenoside Rb₃, chikusetsusaponin IV, chikusetsusaponin IVa, ginsenoside Rd, zingibroside R₁, and calenduloside E in Panacis Majoris Rhizoma. The method demonstrated good linearity within the tested mass concentration range for the eight target components. The relative standard deviations (RSD) of precision, repeatability, and stability were all less than 3.0%. The average recovery rates of ginsenoside Rb₁, ginsenoside Ro, ginsenoside Rb₃, chikusetsusaponin IV, chikusetsusaponin IVa, ginsenoside Rd, zingibroside R₁, and calenduloside E were 102.4%, 103.1%, 97.97%, 99.42%, 102.7%, 102.1%, 95.23% and 100.4%, respectively, with RSD values of 1.0%, 0.98%, 0.81%, 2.3%, 0.81%, 1.8%, 0.96% and 1.8%. The method established in this study allows for rapid and accurate qualitative and quantitative analysis of saponin components in Panacis Majoris Rhizoma.