柴胡中酰化柴胡皂苷的含量测定

Determination of Acyl-Saikosaponin in Radix Bupleuri

目的建立柴胡中酰化柴胡皂苷含量的测定方法。方法采用单因素分析和正交实验设计法,选择氨水浓度、超声时间、超声功率为因素各取3水平,按L9(34)正交表进行实验,优化柴胡中酰化柴胡皂苷的水解方法。采用HPLC测定柴胡皂苷的含量。色谱条件为Wonda Sil C18色谱柱(4.6 mm×250 mm,5μm);流动相:甲醇-水(53∶47);流速:1.0 m L·min-1;检测波长:210 nm。结果优化的柴胡中酰化柴胡皂苷水解方法为浓氨水的浓度为23%,超声25 min,超声的功率为350 W。基于所测定的样品中酰化柴胡皂苷的含量结果,柴胡药材中酰化柴胡皂苷的含量相低于柴胡皂苷,且在柴胡皂苷含量的70%以下。结论该方法简便可靠,可用于柴胡中酰化柴胡皂苷的含量测定。为准确评价柴胡药材质量提供参考。

OBJECTIVE To establish the determination method of acyl-saikosaponin in Radix Bupleuri. METHODS Single factor analysis and orthogonal experimental design were used to optimize the acyl-saikosaponin hydrolysis parameters such as concentration of aqueous ammonia, time of ultrasound-assisted hydrolysis and uhrasound power by L9 （3^4） orthogonal test. An HPLC method was employed to determine the saikosaponins contents, which was carried out on a WondaSil C18 column （4. 6 mm×250mm,5μm） with the mobile phase of methanol-water （53：47） at the flow rate of 1.0 mL · min^-1 and the wavelength of 210 nm. RESULTS The optimized conditions of acyl-saikosaponin hydrolysis were as follows ： the concentration of aqueous ammonia was 23%, the time of ultrasound-assisted hydrolysis was 25 min, and the ultrasound power was 350 W. The content of acyl-saikosaponin in the Radix Bupleuri samples purchased in 12 different drug stores was lower than that of saikosaponin in Radix Bupleuri. CONCLUSION This method is simple and reliable, which can be used for the determination of acyl-saikosaponin in Radix Bupleuri and provides clues for the quality assessment of Radix Bupleuri.