HPLC-DAD法同时测定鹅不食草中5种黄酮类成分的含量

Simultaneous Determination of Five Active Constituents in Centipeda Minima(L.) A. Br. et Aschers. by HPLC-DAD

目的:建立HPLC-DAD法同时测定中药鹅不食草中槲皮素、山奈酚、3-甲氧基槲皮素、芹菜素、蜜橘黄素5种黄酮类活性成分的含量。方法:采用Diamonsil C_(18)色谱柱(200 mm×4.6 mm,5μm),流动相为乙腈(A)-0.1%磷酸水溶液(B),梯度洗脱,流速0.8 ml·min^(-1),检测波长360 nm,柱温30℃。结果:槲皮素、山奈酚、3-甲氧基槲皮素、芹菜素、蜜橘黄素的质量浓度分别在0.002 3~0.093 0μg·μl^(-1)(r=0.999 5)、0.002 2~0.087 0μg·μl^(-1)(r=0.999 6)、0.002 0~0.079 0μg·μl^(-1)(r=0.999 8)、0.000 9~0.037 0μg·μl^(-1)(r=0.999 8)和0.000 8~0.031 0μg·μl^(-1)(r=0.999 9)范围内与峰面积呈良好的线性关系。槲皮素、山奈酚、3-甲氧基槲皮素、芹菜素、蜜橘黄素的平均加样回收率分别为97.66%、98.33%、98.63%、98.40%和98.10%,RSD分别为1.17%、1.16%、1.10%、1.52%和1.36%(n=6)。结论:该方法简便、准确、重复性好,可用于鹅不食草的质量控制。

Objective ： To develop an HPLC-DAD method for the simultaneous determination of five active flavonoids （ quercetin, kaempferol, apigenin, 3-methoxyl-quercetin, nobiletin） in Centipeda minima （L.） A. Br. et Aschers. Methods： The chromatographic separation was performed on a Diamonsil C18 column （200 mm × 4.6 mm,5 μm） with the mobile phase of 0.1% phosphoric acid-acetonitrile with gradient elution at the flow rate of 0.8 ml·min^-1 The detection wavelength was set at 360nm, and the column tempera- ture was maintained at 30℃. Results： Quercetin, kaempferol, 3-methoxyl-quercetin, apigenin and nobiletin was linear within the range of 0.002 3-0.093 0 μg·μl^-1 （r =0.999 5） , 0.002 2-0.087 0 μg·μl^-1 （r =0.999 6） ,0.002 0-0.079 0 μg·μl^-1 （r = 0.999 8） ,0.000 9-0.037 0 μg·μl^-1 （r =0.999 8） and 0.000 8-0.031 0 μg·μl^-1 （r =0.999 9） , respectively. The average recovery was 97.66% （ RSD = 1.17% ） , 98.33% （ RSD = 1.16% ） , 98.63% （ RSD = 1.10% ） , 98.40% （ RSD = 1.52% ） and 98.10% （RSD = 1.36% ）（ n = 6） , respectively. Conclusion： The method is convenient, accurate and reproducible, which can be used for the quality control of Centipeda mirtima （L.） A. Br. et Asehers.