Simultaneous Determination of 10 Flavonoids and Coumarins in Green Aurantii Fructus from Different Origins by HPLC

A method was established for simultaneous determination of 10 flavonoids and coumarins in the green aurantii fructus from different orignis by HPLC. 10 compounds in the sample were untrasonically extracted with 50% (volume fraction) methanol solution. After centrifugation and filtration, the filtrate was analyzed by HPLC. Extend XDB C₁₈ (250 mm×4.6 mm, 5 μm) column was adopted as stationary phase, the mixed solution composed of acetonitrile and 0.1% (volume fraction) formic acid solution with various volume ratios was used as mobile phase in gradient elution, and the target compounds were determined at the detected wavelength of 330 nm. As shown by results, linearity relationships between the mass concentration of 10 compounds (narirutin, naringin, hesperidin, neohesperidin, meranzin hydrate, luteolin, meranzin, nobiletin, hesperetin and aurapten) and respective peak area were kept in the definite ranges, with detection limits in the range of 0.019-0.105 mg·g^-1. The proposed method was applied to the analysis of sample No. S3, RSDs (n=7) of peak areas for 10 compounds in sample solutions were less than 2% in 24 h, and RSDs of peak areas for 6 parallel determination were less than 2.2%. This showed that the method had good stability and repeatability. The spiked recovery test was made at the 3 concentration levels using sample as matrix, giving recoveries in the range of 94.6%-104%. Narirutin, hesperidin, nobiletin and hesperetin were detected in 13 samples, the content of these compounds in sample from different origins varied greatly. The link cluster analysis was made by using the determined results of the characteristic substances of citrus clementine and tangeretin as variables, and results obtained could judge the relationship between the species.