知母甾体总皂苷中知母皂苷BⅡ的鉴别及含量测定

Identification and Determination of Anemarrhenasaponin BⅡ in Total Anemarrhenasaponin Extract by TLC and HPLC-ELSD

目的建立知母甾体总皂苷提取物中知母皂苷BⅡ的TLC鉴别及HPLC含量测定的方法。方法 TLC法:展开剂:正丁醇-冰醋酸-水(4∶1∶5)的上层溶液,显色剂:香草醛硫酸试液,105℃加热至斑点清晰。HPLC-ELSD法:采用Agilent HC-C18(5μm,4.6 mm×250 mm),流动相:甲醇-水梯度洗脱,流速:1 ml.min-1,漂移管温度40℃,载气压力3.5 bar。结果知母皂苷BII成分的TLC分离效果较好;HPLC法测得知母皂苷BII进样量在0.506～10.12μg范围内呈良好的线性关系;平均回收率(n=6)为97.44%,RSD为1.67%。结论该方法简便准确,可用于知母甾体总皂苷提取物的质量控制。

Objective To establish methods for indentification and determination of Anemarrhenasaponin B Ⅱ in Total Anemarrhenasaponin extract by TLC and HPLC - ELSD. Methods The Anemarrhenasaponin B Ⅱ was identified on a silica gel thin layer plate developed with n - butanol - acetic acid - water（4： 1： 5 ） , vanillin - sulfuric acid was used for visualization. The separation of Anemarrhenasaponin B Ⅱ was performed on a Agilent HC - C18 （5 μm,4.6 mm × 250 mm） column of HPLC. The mobile phase was methanol - water and the flow rate was 1ml/min. The detector was ELSD. The temperture of drift tube was 40℃ and the pressure was 3.5bar. Results Anemarrhenasaponin B 1] was well separated by TLC. The linear range of the component was 0.506 × 10.12 μg. The average recovery of Anemarrhenasaponin B 11 was 97.44% with RSD of 1.67%. Conclusion The method is reliable, simple and precise, and it can be used for the determination of total Anemarrhenasaponin extraction.