毛冬青根中4个绿原酸类及4个皂苷类成分的含量测定

目的：建立HPLC法同时测定毛冬青根中绿原酸，异绿原酸A、B、C，毛冬青皂苷A₁、B₁、B₂，冬青素A共8个成分含量。方法：采用Inertsil ODS-4 C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈为流动相A，0.1%磷酸水溶液为流动相B，梯度洗脱，流速为1 mL·min^-1，检测波长327 nm（绿原酸类成分）、210 nm（皂苷类成分）。结果：在上述条件下，绿原酸，异绿原酸B、A、C，毛冬青皂苷B₂、A₁、B₁，冬青素A的质量浓度分别在1.25~12.5、2.96~29.6、3~30、3.45~34.5、13.05~130.5、32.4~324、16.95~169.5、72.8~728 μg·mL^-1范围内与峰面积呈良好的线性关系，平均加样回收率（n=9）均高于95%，RSD均小于3.0%。3批样品中上述8个成分测定结果分别为0.13~0.15、0.23~0.27、0.31~0.37、0.27~0.28、1.24~1.65、2.42~2.86、2.18~2.81、8.29~10.44 mg·g^-1。结论：该方法适用于同时测定毛冬青根中8个成分的含量。

Objective:To establish an HPLC method for simultaneous determination of chlorogenic acid,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,ilexsaponin A₁,ilexsaponin B₁,ilexsaponin B₂ and ilexgenin A in the roots of Ilex pubescens.Methods:HPLC separation was carried on an Inertsil ODS-4 C₁₈ column（4.6 mm×250 mm，5 μm）with the mobile phase consisting of acetonitrile as mobile phase A and 0.1% phosphoric acid as mobile phase B in a program of gradient elution.The flow rate was 1 mL·min^-1 and the detection wavelengths were set at 327 nm for chlorogenic acids and 210 nm for saponins.Results:Under the above conditions,the linear ranges of chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,ilexsaponin B₁,ilexsaponin A₁,ilexsaponin B₁ and ilexgenin A were 1.25-12.5 μg·mL^-1,2.96-29.6 μg·mL^-1,3-30 μg·mL^-1,3.45-34.5 μg·mL^-1,13.05-130.5 μg·mL^-1,32.4-324 μg·mL^-1,16.95-169.5 μg·mL^-1,72.8-728 μg·mL^-1,respectively. The average recoveries(n=9)were above 95.0% with RSD values less than 3.0%.Contents of the analytes in the samples were 0.13-0.15 mg·g^-1,0.23-0.27 mg·g^-1,0.31-0.37 mg·g^-1,0.27-0.28 mg·g^-1,1.24-1.65 mg·g^-1,2.42-2.86 mg·g^-1,2.18-2.81 mg·g^-1,8.29-10.44 mg·g^-1,respectively.Conclusion:The established method was applicable in the determination of chlorogenic acids and saponins in Radix Ilicis Pubescentis.