A novel analytical method based on ultra-performance liquid chromatography-diode array detector (UPLC-DAD), solid-phase extraction (SPE), and quantitative analysis of multiple components by single marker (QAMS) was developed for the first simultaneous determination of 8 flavonoids in Pteridium aquilinum. The combination of QAMS and SPE demonstrated the convenient and economic advantages of the QAMS method while also exhibiting straightforward and cost-efficient advantages in terms of both the time and solvent usage of the SPE. The contents of the 8 flavonoids in P. aquilinum ranged from 0.59 to 697.08 μg/g. There were no substantial differences in the quantification when comparing the QAMS method and the external standard method (ESM). The standard deviations (SDs, %) obtained by QAMS and ESM were all less than 5 %. SPE increased the concentration of flavonoids by 4.18–18.9 times. The limits of detection (LODs) and quantification (LOQs) were in the ranges of 0.01–0.15 and 0.06–0.33 ng/g, respectively. Method validation revealed the linearity of the calibration curves (R2 ≥ 0.9989), with recoveries between 97.02 % and 105 %, and the relative standard deviations (RSD) were all lower than 5 %. In addition, rhoifolin (4), apigenin (6), and amentoflavone (8) were first reported in P. aquilinum. The results suggested that this method was simple, reliable, and feasible for determining the flavonoids in P. aquilinum samples and could be further applied to various foods and herbal medicine samples.
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