目的： 建立1种用HPLC同时测定蒙古黄芪中4个黄酮和2个皂苷含量的方法。 方法： 采用Inertsil ODS-SP（250 mm×4.6 mm，5 μm）色谱柱，以乙腈（A）-水（B）为流动相，梯度洗脱（0～9 min，25%A→45%A；9～20 min，45%A→75%A），黄芪甲苷（S1）、3'-羟基-4-甲氧基异黄酮-7-O-β-D-葡萄糖苷（S2）和黄芪皂苷Ⅱ（S3）的检测波长为205 nm（检测时间段0～9 min），4-甲氧基异黄酮-7-O-β-D-葡萄糖苷（S4）、9，10-二甲基紫檀烷-3-O-β-D-葡萄糖苷（S5）和7-羟基-3'，4'-二甲基-异黄烷醌（S6）的检测波长为245 nm（检测时间段9～20 min）；柱温为40℃。 结果： 在选定的实验操作条件下，S1、S2、S3、S4、S5和S6对照液分别在20.00～100.00 μg·mL-1（r=0.9992）、1.00～10.00 μg·mL-1（r=0.9997）、20.00～100.00 μg·mL-1（r=0.9991）、1.00～10.00 μg·mL-1（r=0.9999）、10.00～60.00 μg·mL-1（r=0.9995）和1.00～10.00 μg·mL-1（r=0.9997）浓度范围内具有良好的线性关系，平均回收率分别为96.5%（RSD=2.0%），98.4%（RSD=1.4%），95.5%（RSD=2.1%）， 98.5%（RSD=1.1%），98.8%（RSD=1.3%），98.7%（RSD=1.1%）。 结论： 采用HPLC在多波长检测条件下可同时测定4个黄酮（2个异黄酮、2个黄烷醇）和2个皂苷的含量，方法简便快速，可为评价该药材质量提供依据。
Objective: To develop an HPLC method for the simultaneous determination of four flavonoids and two saponins. Methods: The gradient elution was performed for the sample separation by using Inertsil ODS-SP columns(250 mm×4.6 mm,5 μm)with acetonitrile(A)-water(B)as the mobile phase.The separation was achieved using the following gradient:0-9 min,25%A→45%A;9-20 min,45%A→75%A,and the column temperature was controlled at 40°C.Astragaloside(S1),3'-hydroxy-4-methoxy isoflavone-7-O-β-D-glucosid(S2)and astragaloside Ⅱ(S3)were detected during 0-9 min with a detection wavelength at 205 nm;4-methoxy isoflavone-7-O-β-D-glucosid(S4),9,10-two methyl pterocarpan-3-O-β-D-glucosid(S5)and 7-hydroxy-3',4'-two methyl isoflavan quinine(S6)were detected during 9-20 min with a detection wavelength at 245 nm. Results: The linear ranges of S1,S2,S3,S4,S5 and S6 were 20.00-100.00 μg·mL-1(r=0.9992),1.00-10.00 μg·mL-1(r=0.9997),20.00-100.00 μg·mL-1(r=0.9991),1.00-10.00 μg·mL-1(r=0.9999),10.00-60.00 μg·mL-1(r=0.9995)and 1.00-10.00 μg·mL-1(r=0.9997),respectively.The average recoveries of S1,S2,S3,S4,S5 and S6 were 96.5%(RSD=2.0%),98.4%(RSD=1.4%),95.5%(RSD=2.1%),98.5%(RSD=1.1%),98.8%(RSD=1.3%),and 98.71%(RSD=1.1%),respectively. Conclusion: The content of four flavonoids(two isofl avones and two flavanols)and two saponins can be simultaneously determined by reversed phase high performance liquid chromatography under the multi wavelength conditions.The established method is simple and rapid,which can provide the basis for the quality control of this medicinal material.
