HPLC-ELSD法同时测定商陆中四种皂苷的含量

Simultaneous Determination of Four Triterpenes in Radix Phytolaccae by HPLC-ELSD

目的:建立HPLC-ELSD法同时测定商陆中商陆皂苷A、商陆皂苷B、商陆皂苷C和商陆皂苷H的含量。方法:采用Diamonsil C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇(A)-0.1%冰乙酸水溶液(B),梯度洗脱(0~20 min,50%A^90%A),流速1.0 mL·min^(-1),用蒸发光散射检测器进行检测,柱温30℃,气体流速2.0 L·min^(-1),漂移管温度50℃。结果:商陆皂苷A、商陆皂苷B、商陆皂苷C和商陆皂苷H分别在0.719~8.628、0.376~4.518、0.258~3.102、0.503~6.036μg范围内,进样量的对数与峰面积对数呈现良好的线性关系(r=0.9992~0.9996),平均加样回收率(n=6)在99.52%~100.17%之间,RSD为1.83%~2.34%。30批商陆样品中商陆皂苷A、商陆皂苷B、商陆皂苷C和商陆皂苷H的含量(n=3)测定结果分别为1.53~4.58、0.16~1.40、0.10~0.61、0.13~4.98 mg·g^(-1)。结论:该方法准确且重复性好,适用于商陆中四种三萜皂苷的含量测定。

To develop the HPLC-ELSD method for simultaneous determination of four triterpenes,including esculentoside A,esculentoside B,esculentoside C,and esculentoside H. Methods: The Diamonsil C_(18) column( 250 mm × 4. 6 mm,5 μm) was used. The mobile phase was methanol( A) and 0. 1% acetic acid( B) with a gradient elution( 0-20 min,50% A-90% A),the flow rate was 1. 0 mL·min^(-1). HPLC-EISD was applied to determine the content,the column temperature was 30℃,gas flow rate was 2. 0 L·min^(-1),and drift tube temperature was 50℃. Results: The calibration curves of esculentoside A,esculentoside B,esculentoside C and esculentoside H showed good liner relationship between logarithm of the peak area and logarithm of the content of each compound over 0. 719-8. 628,0. 376-4. 518,0. 258-3. 102,and 0. 503-6. 036 μg respectively( r = 0. 9992-0. 9996). The average recoveries( n = 6)were in the ranges of 99. 52% -100. 17% and RSD was in the ranges of 1. 83% -2. 34% respectively. The contents in 30 batches of samples of Radix Phytolaccae were 1. 53-4. 58 mg·g^(-1) for esculentoside A,0. 16-1. 40 mg·g^(-1) for esculentoside B,0. 10-0. 61 mg·g^(-1) for esculentoside C,and 0. 13-4. 98 mg·g^(-1) for esculentoside H. Conclusion: The method is precise and repeatable,which can be used to determine the contents of four triterpenoid saponins in Radix Phytolaccae.