HPLC法同时测定枳壳中伞形花内酯等9种成分的量

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

目的建立同时测定枳壳中伞形花内酯、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、川陈皮素、3,5,6,7,8,(3,4)-七甲氧基黄酮、橘红素、橙皮油素的HPLC方法,测定和比较不同周长大小枳壳中的9种化学成分的量。方法采用Diamonsil C18(250 mm×4.6 mm,5μm),流动相为甲醇(A)-水(用磷酸调p H至3.0);梯度洗脱为0~25 min,30%~50%A;25~35 min,50%~70%A;35~40 min,70%~75%A;40~55 min,75%~100%A。体积流量为1.0 mL/min;柱温30℃;检测波长320 nm;进样量20μL。结果伞形花内酯、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、川陈皮素、3,5,6,7,8,(3,4)-七甲氧基黄酮、橘红素和橙皮油素分别在0.002 18~0.07μg(r=0.999 8)、0.021 8~0.70μg(r=0.999 9)、0.242 5~7.76μg(r=0.999 8)、0.024 38~0.78μg(r=0.999 4)、0.523 76~16.76μg(r=0.999 3)、0.004 13~0.132μg(r=0.999 6)、0.003 13~0.10μg(r=0.999 3)、0.002 75~0.088μg(r=0.999 6)和0.000 93~0.03μg(r=0.999 3)呈良好的线性关系;其平均回收率(n=6)分别为98.50%、98.80%、99.51%、98.43%、99.64%、99.21%、100.03%、98.75%和101.11%。结论该法简便,重复性好,可用于枳壳中伞形花内酯、3,5,6,7,8,(3,4)-七甲氧基黄酮等成分的含量测定。

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 p H 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.