Analysis of main active components in Desmodii Styraciflii Herba by HPLC-Q- TOF-MS and HPLC-DAD / 中草药

ABSTRACT

Objective: This study is aimed to establish a high performance liquid chromatography coupled with quadrupole-time-of- flight mass spectrometric (HPLC-Q-TOF-MS) method for analyzing the main active components in Desmodii Styraciflii Herba, and to establish an HPLC-DAD method for simultaneously determining its seven flavonoids compounds (schaftoside, isoschaftoside, vicenin-2, isoorientin, isovitexin, luteolin, and apigenin). Methods: The analysis was performed on a Phenomenex Kinetex C18 column with a gradient elution of methanol-0.2% aqueous formic acid at the flow rate of 1.0 mL/min. The column temperature was 40 oC. The Q-TOF-MS discriminant analysis was performed under negative electrospray ion mode and the split ratio was 1∶1. Quantitative analysis of seven compounds in Desmodii Styraciflii Herba was carried by HPLC-DAD. The determination wavelength was at 272 nm. Results: Forty-one main active constituents were separated and identified by HPLC-Q-TOF-MS, including 37 flavonoids compounds and four phenolic acids. A HPLC-DAD method was successfully developed for determining seven flavonoids compounds. The standard curves for all seven compounds showed good linearity with correlation coefficients higher than 0.999 0 within the test ranges. The average recoveries (n = 6) were between 96.7%-102.4%, and RSD ≤ 4.94%. Conclusion: Analyzing the main active components provides a significant guidance for the substance basis research of Desmodii Styraciflii Herba. The HPLC-DAD method for simultaneously determining seven flavonoids compounds is accurate, reproducible, and provides the basis for the quality control of Desmodii Styraciflii Herba.