HPLC-ELSD法同时测定河北产道地药材不同物候期知母中知母皂苷AⅢ和知母皂苷BⅡ的含量

Simultaneous determination of timosaponin AⅢ and timosaponin BⅡ in Rhizoma Anemarrhenae from Hebei in the different phenological periods by HPLC-ELSD

目的:建立同时测定知母药材中知母皂苷AⅢ和知母皂苷BⅡ含量的高效液相色谱-蒸发光散射检测法。研究不同物候期知母道地药材中2种皂苷类成分的积累动态,为最佳采收期的制定、品质评价提供参考。方法:采用Thermo C18色谱柱(4.6mm×250mm,5μm),乙腈-水梯度洗脱,流速:1mL/min,柱温:25℃。ELSD检测器雾化器流速:2.8L/min,漂移管温度:100℃。结果:知母皂苷AⅢ在0.6950-4.1700μg范围内(r=0.9993),知母皂苷BⅡ在1.0175-6.1050μg范围内(r=0.9996),进样质量的对数值与峰面积的对数值呈良好的线性关系。知母皂苷AⅢ平均回收率为99.88%,RSD=2.34%;知母皂苷BⅡ平均回收率为103.66%,RSD=0.99%。结论:该方法简便、准确、可靠,可同时测定知母药材中2种皂苷类成分的含量。不同物候期知母道地药材中两种皂苷的含量存在显著差异。

Objective： To establish a HPLC-ELSD method for simultaneous determination of two compounds including timosaponin Am and timosaponin BⅡ in Rhizoma Anemarrhenae. And study the accumulation trends of timosaponin Am and timosaponin B Ⅱ in Rhizoma Anemarrhenae from Hebei in the different phenological periods for defining the best harvest time and quality evaluation of Rhizoma Anemarrhenae. Methods： Two compounds were simultaneously determined by HPLC-ELSD with Thermo C18 （4.6mm×250mm, 5μm） column. The mobile phase was：consisted of acetonitrile （A） and water （B） with gradient elution at the flow rate of lmL/min. The column temperature was set at 25℃. The ELSD conditions were as follows： flow rate of gas 2.8L/min and temperature of drifttube 100℃. Results： The linear ranges of timosaponin A Ⅲ and timosaponin B Ⅱ were 0.6950-4.1700pg （r=0.9993）, 1.0175-6.1050μg （r=-0.9996）. The average recoveries of the method were 99.88%, RSD=2.34%, 103.66%, RSD=0.99%, respectively. Conclusion： The method is simple, reliable and accurate, and has been successfully used to the quantification of two compounds above mentioned in Rhizoma Anemarrhenae. And the quantifications of two compounds of Rhizoma Anemarrhenae in different phenological periods are significant different.