HPLC法同时测定杜仲叶中6个黄酮类成分的含量

Simultaneous determination of six flavonoids in leaves of Eucommia ulmoides Oliv. by HPLC

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（250mm×4.6 mm,5μm）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、0.043~0.32μg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 C18column（ 250 mm × 4. 6 mm,5μm） 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. 0mL·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% and102. 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.