Rapid extraction and analysis method for the simultaneous determination of 21 bioflavonoids in Siegesbeckia pubescens Makino.

A novel and rapid microwave extraction and ultra high performance liquid chromatography coupled with a triple quadrupole-linear ion trap mass spectrometry method was developed and validated for the determination of 21 bioflavonoids in Siegesbeckia pubescens Makino. The optimal conditions for the extraction of flavonoids from Siegesbeckia pubescens Makino involved the use of methanol as the extraction solvent, a microwave temperature of 70°C, an extraction time of 11 min, and a solvent-to-solid ratio of 40 mL/g. Chromatographic separation was achieved on a Waters ACQUITY UPLC BEH C18 column(100 × 2.1 mm, 1.7 μm) with a gradient mobile phase (A: 0.3% v/v aqueous formic acid and B: acetonitrile) at a flow rate of 0.25 mL/min. All calibration curves showed good linearity (r > 0.999) within the test ranges. The method developed was validated with acceptable sensitivity, intra- and interday precision, reproducibility, and extraction recoveries. The validated method was successfully applied to determine the contents of 21 bioflavonoids in Siegesbeckia pubescens Makino from different sources.