Objective：To establish a high-performance liquid chromatography-mass spectrum method for determination of panaxsaponin Rg1, icariin, psoralen and isopsoralen in Hongzhu Callrhinus pills. Methods：An Agilent Tc-C18 (4.6 mm×250 mm，5 μm) column was used for the separation, and gradiently eluted by 0.1% acetic acid－acetonitrile with flow rate of 1.0 mL·min-1. AP-ESI with positive ion mode and the selected ion monitoring were used. The ion mass spectrum (m/z) of 209, 321, 677 and 823 were selected to quantify panaxsaponin Rg1, icariin, psoralen and isopsoralen, respectively, with lorazepam as an internal standard. Results：The liner range of the calibration curves for panaxsaponin Rg1, icariin, psoralen and isopsoralen were 2.02-12.12, 2.0-12.0, 0.53-1.86, 0.4-1.4 μg·mL-1 respectively. The contents of panaxsaponin Rg1, icariin, psoralen and isopsoralen per Hongzhu Callrhinus pill were(232.6±0.64)μg，(669.0±14.51)μg，(564±34.71)μg and (428±20.17)μg. Conclusion：The established method is convenient, rapid and accurate, which can be used to determine four components in Hongzhu Callrhinus pills to control its qualities, or to investigate its pharmacokinetic properties in human or animals.
