Separation and quantification of flavonoid in rhododendron anthopogonoides maxim by high performance liquid chromatography

A high-performance liquid chromatographic (HPLC) method has been developed for separation and quantitative analysis of flavonoid aglycones in Rhododendron anthopogonosides Maxim. Flavonoids in their bound forms were hydrolyzed with acid before HPLC analysis. Analytical samples were pretreated by solid-phase extraction on C18 reversed-phase cartridges. Optimum separation on a 4.6 mm × 250 mm i.d. C18 column was achieved by use of a 52:48 (v/v) mixture of methanol and an aqueous solution of 10 mm citric acid and 1 mm sodium dodecyl sulfate as mobile phase. The flow rate was 1.0 mL min−1 and the detection wavelength 360 nm. Five flavonoids, myricetin, quercetin, luteolin, kaempferol, and isorhamnetin, were separated with high resolution without use of gradient elution. The method was successfully used for efficient qualitycontrol analysis by quantifying flavonoids in R. anthopogonosides. Repeatability tests showed that intra-day and inter-day RSD was <10%. LOD of the five flavonoids were <0.85 μg mL−1. Recovery ranged from 90.2 to 112.5%, with RSD <11.1%.