RP-HPLC法同时测定雪松松针中杨梅素、槲皮素和山柰酚的含量

目的:建立同时测定雪松松针中3个黄酮类化合物杨梅素、槲皮素和山柰酚含量的反相高效液相色谱法。方法:样品用50倍60%甲醇回流提取1 h,提取液加10%盐酸水解1 h。采用TC-C₁₈(250 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速1.0 mL·min^-1,检测波长360 nm,柱温30℃。结果:雪松松针中杨梅素、槲皮素和山柰酚3个黄酮类化合物浓度在0.4~8.0 μg·mL^-1、0.5~9.5 μg·mL^-1和0.8~15.2 μg·mL^-1范围内呈良好线性关系,相关系数在0.9998~0.9999之间;加样回收率(n=9)分别为98.4%,97.5%,98.9%。结论:该方法简单、快速、高效,可用于雪松松针中黄酮类化合物的测定。

Objective:To develop a RP-HPLC method for the simultaneous determination of three flavonoids(myricetin,quercetin,kaempferol) in pine needles of Cedrus deodara . Methods:The samples were pretreated,including reflux extraction with 60% methanol for 1 h(with the ratio of liquor to material of 50:1),and then extracting solution hydrolysis with 10% hydrochloric acid for another 1 h.Agilent TC-C₁₈ (250 mm×4.6 mm,5 μm) column was adopted;The mobile phase was acetonitrile-0.1% phosphoric acid solution with the gradient elution at a flow rate of 1.0 mL·min^-1;The column temperature was 30℃ and the detection wavelength was 360 nm. Results:Good linear correlations were found in the ranges of 0.4-8.0 μg·mL^-1 for myricetin,0.5-9.5 μg·mL^-1 for quercetin and 0.8-15.2 μg·mL^-1 for kaempferol,respectively.The correlation coefficients were between 0.9998-0.9999.The average recoveries (n=9) of the three flavonoids were 98.4%,97.5% and 98.9%,respectively. Conclusions:The method is simple,rapid and efficient,which is applicable for determination of flavonoids in pine needles of C.deodara .