Simultaneous determination of six bioactive flavonoids in Citri Reticulatae Pericarpium by rapid resolution liquid chromatography coupled with triple quadrupole electrospray tandem mass spectrometry

A rapid resolution liquid chromatography/electrospray ionisation tandem mass spectrometry (RRLC–ESI-MSn) method has been firstly developed and validated for simultaneous determination of six bioactive flavonoids in Citri Reticulatae Pericarpium (CRP). The antiproliferative activities of the six flavonoids in CRP, namely naringin, hesperidin, nobiletin, 3,5,6,7,8,3′,4′-heptamethoxyflavone, tangeretin and 5-hydroxy-6,7,8,3′,4′-pentamethoxyflavone, were evaluated and compared by Cell Counting Kit-8 Assay. Quantification was carried out on an Agilent triple quadrupole LC–MS system using multiple reaction monitoring mode. The established method was successfully applied for determination of the six flavonoids in samples collected from different regions in China. Compared with the reported analytical methods, the RRLC–ESI-MSn method is powerful in quantitative analysis of multi-component in terms of time savings and sensitivity. Hierarchical cluster analysis (HCA) was also performed to differentiate and classify the samples based on the contents of the six characteristic flavonoids. The HCA results indicated that Citrus reticulata ‘Chachi’ samples could be easily distinguished from other CRP samples. The developed RRLC–ESI-MSn method combined with HCA might be utilised as a quality control method for CRP.