Simultaneous determination of 6 steroidal saponins in Paris Polyphylla by accelerated solvent extraction-HPLC-TOF/MS / 第二军医大学学报

ABSTRACT

Objective To 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. Methods The extraction was performed by ASE under the following conditions: extracting solvent 70% ethanol; extracting temperature 120°C; 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°C, nebulizer gas pressure was 275. 8 kPa (40 psi), drying gas flow rate was 10 L/min, and gas temperature was 350°C. Results The 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. Conclusion The 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.