HPLC法同时测定红参中6种稀有人参皂苷的含量

Determination of 6 rare ginsenosides in red ginseng by HPLC

目的:采用高效液相色谱法同时测定红参中6种稀有人参皂苷的含量。方法:以人参皂苷为对照品,采用Agilent ZORBAX SB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-水,梯度洗脱,流速1.0 mL/min;检测波长203 nm;柱温30℃;进样量10μL。结果:6种稀有人参皂苷的色谱峰分离良好,Rk_(3)、Rh_(4)、20-(S)-Rg_(3)、20-(R)-Rg_(3)、Rk_(1)、Rg_(5)质量浓度分别在8.7~259.8μg/mL(r=0.9990)、12.7~240.0μg/mL(r=0.9990)、9.3~185.8μg/mL(r=0.9995)、16.6~198.0μg/mL(r=0.9996)、13.9~220.7μg/mL(r=0.9992)和10.3~164.5μg/mL(r=0.9993)范围内线性关系良好,方法的平均加样回收率分别为96.90%(RSD=1.89%)、97.37%(RSD=1.87%)、101.95%(RSD=3.29%)、105.68%(RSD=2.83%)、102.38%(RSD=1.76%)和92.84%(RSD=4.00%)。10批市售红参药材均含6种稀有人参皂苷且含量均较低,各批次间含量差异较大。结论:建立的含量测定方法准确灵敏,稳定性和重复性好,可为红参的质量研究提供依据。

Objective:To establish the HPLC method for the determination of six rare ginsenosides in red ginseng.Methods:With ginsenoside as reference substances,HPLC analysis was performed on an Agilent ZORBAX SB-C_(18)column(4.6 mm×250 mm,5μm).The mobile phase was acetonitrile-water with gradient elution.The flow rate was 1.0 mL/min,the detection wavelength was 203 nm and the column temperature was 30℃,the quantity of sampling was 10μL.Results:The chromatographic peaks of six rare ginsenosides were well separated.The linear ranges of ginsenoside Rk 3,ginsenoside Rh_(4),20-(S)-ginsenoside Rg_(3),20-(R)-ginsenoside Rg_(3),ginsenoside Rk_(1)and ginsenoside Rg 5 were 8.7-259.8μg/mL(r=0.9990),12.7-240.0μg/mL(r=0.9990),9.3-185.8μg/mL(r=0.9995),16.6-198.0μg/mL(r=0.9996),13.9-220.7μg/mL(r=0.9992)and 10.3-164.5μg/mL(r=0.9993),respectively.The average recovery rate were 96.90%(RSD=1.89%),97.37%(RSD=1.87%),101.95%(RSD=3.29%),105.68%(RSD=2.83%),102.38%(RSD=1.76%),92.84%(RSD=4.00%),respectively.Ten batches of red ginseng all contain 6 kinds of rare ginsenosides and their contents were low,and the contents vary greatly among batches.Conclusion:The established content determination method is accurate,sensitive,stable and reproducible,which could provide a basis for the quality study of red ginseng.