Simultaneous quantitative determination of nine active compounds from Aurantii Fructus by HPLC / 中草药

ABSTRACT

Objective: To establish an HPLC method for simultaneous determination of 7-hydroxycoumarin, narirutin, naringin, hesperidin, neohesperidin, nobiletin, 3,5,6,7,8,(3,4)-heptamethoxyflavone, tangeretin, and auraptene from Aurantii Fructus, determinate and compare the content of various chemical components from Aurantii Fructus with different perimeters. Methods: The HPLC system consisted of a diamon C18 (250 mm × 4.6 mm, 5 μm), the mobile phase was methanol (A)-water (adjusted pH 3.0 with phosphorous acid). The gradient elution conditions 0-25 min, 30%-50% A; 25-35 min, 50%-70% A; 35-40 min, 70%-75% A; 40-55 min, 75%-100% A. The flow rate was 1.0 mL/min and the column temperature was 30 ℃. The detection wavelength was 320 nm and the injection volume was 20 μL. Results: The calibration curves of 7-hydroxycoumarin, narirutin, naringin, hesperidin, neohesperidin, 3,5,6,7,8,(3,4)-heptamethoxyflavone, nobiletin, tangeretin, and auraptene had good linear relationship in the ranges of 0.002 18-0.07 μg (r = 0.999 8), 0.021 8-0.70 μg (r = 0.999 9), 0.242 5-7.76 μg (r = 0.999 8), 0.024 38-0.78 μg (r = 0.999 4), 0.523 76-16.76 μg (r = 0.999 3), 0.003 13-0.10 μg (r = 0.999 3), 0.004 13-0.132 μg (r = 0.999 6), 0.002 75-0.088 μg (r = 0.999 6), and 0.000 93-0.03 μg (r = 0.999 3); And the average recoveries (n = 6) of the nine components were 98.50%, 98.80%, 99.51%, 98.43%, 99.64%, 99.21%, 100.03%, 98.75%, and 101.11%, respectively. Conclusion: This method can be applied to determinating the components from Aurantii Fructus, including 7-hydroxycoumarin and 3,5,6,7,8,(3,4)-heptamethoxyflavone, etc.