HPLC法同时测定中亚鸢尾中3种抗炎活性成分的含量

Simultaneous Determination of Three Anti-inflammatory Active Ingredients in Iris Bloudowii by HPLC

目的:建立HPLC法同时测定中亚鸢尾中3种具有抗炎活性的黄酮类成分4'-羟基-5,3'-二甲氧基-6,7-亚甲二氧基异黄酮-4'-O-β-D-吡喃葡萄糖苷、尼鸢尾黄素-4'-O-β-D-吡喃葡萄糖苷、4'-羟基-5,3'-二甲氧基-6,7-亚甲二氧基异黄酮含量。方法:色谱柱为HC-C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.3%磷酸溶液,流速:1.0 ml·min^-1,检测波长:265 nm,柱温:25℃,进样量:5μl。结果:4'-羟基-5,3'-二甲氧基-6,7-亚甲二氧基异黄酮-4'-O-β-D-吡喃葡萄糖苷、尼鸢尾黄素-4'-O-β-D-吡喃葡萄糖苷、4'-羟基-5,3'-二甲氧基-6,7-亚甲二氧基异黄酮质量浓度分别在5.25~84.00μg·ml^-1(r=0.9995)、3.25~52.00μg·ml-1(r=0.9996)和3.25~52.00μg·ml^-1(r=0.9998)的范围内与峰面积呈良好的线性关系;平均加样回收率分别为98.07%(RSD=1.58%)、98.65%(RSD=2.17%)和97.21%(RSD=3.46%)(n=9)。结论:该方法准确度好,精密度高,可用于中亚鸢尾中3种黄酮类成分的同时测定。

Objective:To establish an HPLC method for the simultaneous determination of three flavonoids with anti-inflammatory activity in Iris bloudowii.Methods:The method was performed on an HC-C18 column(250 mm×4.6 mm,5μm)with the mobile phase consisting of acetonitrile-0.3%phosphoric acid solution with gradient elution at the flow rate of 1.0 ml·min^-1,the detection wavelength was 265 nm,the column temperature was 25℃and the sample size was 5μl.Results:The linear range of nigricanin-4'-O-β-D-glucoside,nigricin-4'-O-β-D-glucoside and nigricanin was 5.25-84.00μg·ml^-1(r=0.9995),3.25-52.00μg·ml^-1(r=0.9996)and 3.25-52.00μg·ml^-1(r=0.9998),respectively.The average recovery was 98.07%(RSD=1.58%),98.65%(RSD=2.17%)and 97.21%(RSD=3.46%)(n=9),respectively.Conclusion:The method is accurate with high precision,which can be applied to the simultaneous determination of three flavonoids in Iris bloudowii.