Qualitative and Quantitative Analysis of Flavonoids in <i>Humulus scandens</i> (Lour.)Merr. by UPLC-TQD and HPLC

Abstract

To qualify and quantify the flavonoids existing in Humulus scandens (Lour.)Merr., a Ultra performance liquid chromatography triple-quadrupole tandem mass spectrometer (UPLC-TQD) method was applied to preliminarily analyze the flavonoids exiting in Humulus. And a High performance liquid chromatography (HPLC) method was used to analyze the flavonoids existing in Humulus. Flavonoids in methanol extracts of Humulus were cleaned up by a C18 cartridge, separated by a PLASMA ODS column(5μm,250×4.6mm), with a gradient elution of methanol and 0.1% formic acid at a flow rate of 1 mL•min-1 . Analytes were detected with ultraviolet detector (UV) at 280nm. Flavonoids, including Epicatechin, Luteolin-7-O-Glucoside, Vitexin or Apigenin-7-glucoside, Morin hydrate or Quercetin dehydrate and Luteolin were found in Humulus according to the UPLC-TQD. Results of HPLC analysis showed the linear ranges of 11 flavonoids were 0.1mg•L-1-20mg•L-1 with the correlation coefficient (r) all above 0.9909. Limits of detection (signal/ noise=3) was 100Superscript text μg•L-1. The average recoveries of all the compounds spiked at four levels of 200μg•L-1, 500μg•L-1, 1 mg•L-1 and 1.5 mg•L-1 were in the ranges of 70.8%-112.1%, 74.6%-116.6%, 73.1%-110.6%, 73.7%-99.9%, with the corresponding relative standard deviation (RSD, n=6) of 2.5%-5.8%, 0.8%-4.3%, 0.8%-3.7%, 1.7%-3.8%, respectively. This method ,which was proved to be precise, sensitive and reliable, could be applied to analyze flavonoids in leaves.