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℃;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℃.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.
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