ObjectiveTo introduce an accelerated solvent extraction (ASE) -HPLC-TOF/MS method for simultaneous determination of Polyphyllin Ⅰ, Polyphyllin Ⅱ, Polyphyllin Ⅵ, Polyphyllin Ⅶ, Dioscin and Gracillin in Paris Polyphylla. MethodsThe extraction was performed by ASE under the following conditions: extracting solvent 70% ethanol; extracting temperature 120℃; pressure 1.17 MPa (1 700 psi); and static time 6 min. The separation was carried out on a MGC18 column (3.0 mm×100 mm, 3.0 μm). Elution was done with a linear gradient mobile phase system consisting of 0.1% formic acid (V/V) water and acetonitrile. The flow rate was set at 0.8 ml/min, the sample injection volume was 3 μl, the column temperature was kept constant at 25℃, nebulizer gas pressure was 275.8 kPa (40 psi), drying gas flow rate was 10 L/min, and gas temperature was 350℃. ResultsThe calibration curves for Polyphyllin Ⅰ, Polyphyllin Ⅱ, Polyphyllin Ⅵ, Polyphyllin Ⅶ, Dioscin and Gracillin were linear within the range of 0.500 0-50.00 μg/ml (r=0.999 8), 0.422 3-42.23 μg/ml (r=0.999 7), 0.612 0-61.20 μg/ml (r=0.999 9), 0.714 0-71.40 μg/ml (r=0.999 5), 0.448 0-44.80 μg/ml (r=0.999 9) and 0.436 0-43.60 μg/ml (r=0.9997), respectively; and the average recoveries (n=6) were 98.9%, 98.0%, 102.5%, 101.9%, 103.1% and 97.9%, respectively. ConclusionThe introduced method is solvent-saving and time-saving; it can also be used for high-throughput analysis and determination of steroidal saponins in Paris Polyphylla.