HPLC法同时测定杜仲叶中6个黄酮类成分的含量

目的：建立高效液相色谱法同时测定杜仲叶中6个黄酮类成分的含量。方法：采用Thermo BDS HPERSIL C₁₈（250 mm×4.6 mm，5 μm）色谱柱，以甲醇为流动相A，以1%磷酸溶液为流动相B，梯度洗脱（0~50 min，28%A→35%A，72%B→65%B；50~65 min，35%A→60%A，65%B→40%B），流速1.0 mL·min^-1，检测波长360 nm，柱温25 ℃。结果：槲皮素-3-O-葡萄糖（1→2）葡萄糖苷、槲皮素-3-O-阿拉伯糖（1→2）葡萄糖苷、槲皮素-3-O-葡萄糖苷、异鼠李素-3-O-β-D-葡萄糖苷、紫云英苷、槲皮素进样量分别在0.11~0.79、0.21~1.58、1.44~10.76、0.055~0.41、0.055~0.41、0.043~0.32 μg范围内线性关系良好；6个黄酮类化合物的平均回收率分别为101.4%、100.8%、101.9%、99.1%、102.8%、102.0%。结论：建立的方法符合方法学验证要求，可用于杜仲叶中槲皮素、槲皮素苷、异鼠李素苷、紫云英苷等6个黄酮成分的含量测定。

Objective: To establish an HPLC method for simultaneous determination of the contents of 6 flavonoids in the leaves of Eucommia ulmoides Oliv.. Methods: Thermo BDS HPERSIL C₁₈ column (250 mm×4.6 mm,5 μm) was adopted;the mobile phase was methanol(A)-1% phosphoric acid solution(B) with gradient elution(0-50 min,28%A→35%A,72%B→65%B;50-65 min,35%A→60%A,65%B→40%B) at a flow rate of 1.0 mL·min^-1.UV detection was set at 360 nm,and the temperature of column was kept at 25 ℃. Results: The contents of quercetin-3-O-glucose (1→2) glucoside,quercetin-3-O-arabinose (1→2) glucoside,quercetin-3-O-glucoside,isorhamnetin-3-O-β-D-glucoside,astragalin and quercetin had good linear relationship in the ranges of 0.11-0.79,0.21-1.58,1.44-10.76,0.055-0.41,0.055-0.41 and 0.043-0.32 μg,respectively,and the average recoveries of the 6 flavonoids were 101.4%,100.8%,101.9%,99.1%,102.8% and 102.0%,respectively. Conclusion: This method conforms to method validation requirements,and it can be used for the content determination of 6 flavonoid components in the Eucommia ulmoides leaves samples,such as quercetin,quercetin glycoside,isorhamnetin glycosides,astragalin.