Simultaneous Determination of Five Bioactive Flavonoids in Pericarpium Citri Reticulatae from China by High-Performance Liquid Chromatography with Dual Wavelength Detection

To evaluate the quality of Pericarpium Citri Reticulatae from China, a new, simple, and accurate method involving high-performance liquid chromatography with dual wavelength detection was developed for the simultaneous determination of five bioactive flavonoids, hesperidin, nobiletin, 3,5,6,7,8,3',4'-heptamethoxyflavone, tangeretin, and 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone for the first time. Under the optimal condition, analysis was performed on a Dikma Diamonsil C-18 column and gradient elution with a solvent system of acetonitrile and water at a flow rate of 1.0 mL/min and detected at 283 and 330 nm, respectively. All five calibration curves exhibited good linearity (R-2 > 0.9997). The relative standard deviation values for intra- and interday precision were less than 5% with accuracies between 95.16% and 104.71%. The recoveries were in the range of 96.91-103.20%. The established method was successfully applied to determine above five flavonoids in 32 samples collected from different districts of China, and the results demonstrated that the method may be used as strong research tools for quality control of Pericarpium Citri Reticulatae and chemotaxonomic investigation in botanical sciences of Citrus species.