Quantitative Analysis Study of Polyphyllin by Ultrasound-Assisted Eutectic Solvent Extraction Combined with UHPLC-MS/MS

Paris polyphylla (Chonglou), a medicinal herb documented in Shennong’s Classic of Materia Medica and a key component of formulas such as Yunnan Baiyao, is a rare and endangered plant prized for its bioactive steroidal saponins, notably polyphyllin I (PPI) and II (PPII). However, its pharmacological potential is hampered by inefficient extraction and unreliable compound identification. Herein, we developed a sustainable and efficient extraction strategy using ultrasound-assisted deep eutectic solvents (DES), optimized via an L₉(3⁴) orthogonal experimental design. Extraction efficiencies across the seven Paris species ranged from 2.04% to 16.51%, achieved by systematically optimizing key parameters such as the choline chloride-to-ethanol molar ratio (1:1.8), material-to-liquid ratio (1:20 g mL^-1), and extraction time (100 min). By ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis, PPI and PPII were quantified using specific retention times and characteristic fragment ions, revealing content ranges of 3.282–21.452 mg g^-1 and 4.201–17.975 mg g^-1, respectively. This methodology provides a robust platform for quality control and standardization of Paris-derived medicines, while paving the way for sustainable utilization and in-depth study of its steroidal saponins.