Simultaneous determination of ten constituents in the Chinese medicinal preparation fufangxingxiangtu' erfeng capsules by ultra performance liquid chromatography with tandem mass spectrometry

Abstract

Using caffeic acid and icariin as internal standards, a method for the simultaneous determination of protocatechuic acid, protocatechuic aldehyde, chlorogenic acid, scutellarin, isochlorogenic acid C, baicalin, luteolin, apigenin, atractylenolide III and atractylenolide I in Fufangxingxiangtu' erfeng capsules were established by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). The separation was performed on a ZORBAX RRHD Eclipse Plus C18 column (50 mm x 2.1 mm, 1.8 microm) by using water containing 0.3% formic acid and methanol as mobile phases with the gradient elution at a flow rate of 0.3 mL/min. The analytes were detected by a tandem mass spectrometer in the multiple reaction monitoring (MRM) mode via the switching of positive electrospray ionization (ESI(+)) and negative electrospray ionization (ESI(-)). Under optimum conditions, the calibration curves were linear in the range of 0.003 00-24.0 mg/L for protocatechuic acid, 0.017 0-2.00 mg/L for protocatechuic aldehyde, 0.015 0-30.0 mg/L for chlorogenic acid, 0.004 00-30.0 mg/L for scutellarin, 0.010 5-24.0 mg/L for isochlorogenic acid C, 0.003 00-30.0 mg/L for baicalin, 0.003 00-5.0 mg/L for luteolin, 0.006 00-1.50 mg/L for apigenin, 0.001 50-4.00 mg/L for atractylenolide III, and 0.000 600-0.900 mg/L for atractylenolide I with the detection limits of 1.0, 11, 5.0, 1.5, 3.5, 1.0, 1.0, 2.0, 0.50, 0.20 microg/L, respectively. The average recoveries of the ten effective components were between 92.5% and 106% with all relative standard deviations not more than 3.2%. The developed method was rapid, simple, accurate, reproducible, and suitable for the quality control of the Fufangxingxiangtu' erfeng capsules.