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目的:建立测定照山白中黄酮类化合物含量的高效液相色谱(HPLC)方法。方法:85℃下用无水乙醇采用索氏提取装置制备粗黄酮,经过旋转蒸发仪浓缩成浸膏,用高效液相色谱法进行含量测定。采用WatersXTerraMS C_(19)色谱柱(1.5 mm×100 mm,3.5μm);流动相:甲醇-0.2%磷酸水溶液;检测波长:340～380nm;流速:0.2 mL·min~(-1);柱温:室温;自动进样。结果:照山白中黄酮类化合物芦丁,槲皮素,山奈酚在0.0203～162.5μg·mL~(-1)范围内进样量与峰面积呈良好线性关系,其线性回归方程分别为y=2318.3x-240898,r=0.9999;y=5038.1x+143452,r=0.9999;y=2953.1x+414874,r=0.9999。方法学考察表明,黄酮类的最低检测限为0.083μg·mL~(-1),加样回收率为96.60%,98.7%,97.5%,RSD为1.09%,1.20%,1.16%(n=3)。
Objective: To establish a high performance liquid chromatography (HPLC) method for the determination of flavonoids in Zhushan White. Methods: The crude flavonoids were prepared by Soxhlet extraction with anhydrous ethanol at 85 ℃. The crude flavonoids were concentrated by rotary evaporator and the contents were determined by high performance liquid chromatography (HPLC). The mobile phase was methanol-0.2% phosphoric acid aqueous solution, the detection wavelength was 340-380 nm, the flow rate was 0.2 mL · min -1, the column temperature was 1.5 × 100 mm, : Room temperature; automatic injection. Results: The linear regression equations of rutin, quercetin and kaempferol in the range of 0.0203 ~ 162.5μg · mL ~ (-1) showed a good linear relationship with the peak area. The linear regression equations were y = 2318.3x-240898, r = 0.9999; y = 5038.1x + 143452, r = 0.9999; y = 2953.1x + 414874, r = 0.9999. The methodological study showed that the minimum detectable limit of flavonoids was 0.083μg · mL -1, the recoveries were 96.60%, 98.7%, 97.5%, RSD 1.09%, 1.20%, 1.16% (n = 3) ).