高效液相色谱-蒸发光散射检测器法同时测定化瘀祛斑胶囊中4种柴胡皂苷含量

Simultaneous Determination of Four Saikosaponins in Huayu Quban Capsules by HPLC-ELSD Method

目的建立同时测定化瘀祛斑胶囊中柴胡皂苷a,b1,b2,d的高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法。方法色谱柱采用Agilent Zorbax Bonus-RP C18柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)-水(B),梯度洗脱,流速为1.0 m L/min,柱温为30℃,漂移管温度为42℃,载气为氮气(流速为2.5 m L/min)。结果样品中柴胡皂苷a、柴胡皂苷b2、柴胡皂苷b1和柴胡皂苷d进样量分别在0.0821~3.2080μg(r=0.9985),0.2673~10.4400μg(r=0.9998),0.0835~3.2600μg(r=0.9993),0.0909~3.5520μg(r=0.9997)范围内与峰面积平均值的对数值呈良好线性关系;平均回收率分别为101.61%,99.64%,95.36%,95.26%,RSD均小于2%(n=6)。结论该方法操作简单,准确度高,专属性强,可用于化瘀祛斑胶囊中4种柴胡皂苷的含量测定。

Objective To establish a method for simultaneous determination of four saikosaponins a,b1,b2,and d in Huayu Quban Capsules by high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD)method.Methods Agilent Zorbax Bonus-RP C18(250 mm×4.6 mm,5μm)was performed for separation with a gradient program of acetonitrile(A)-water(B)as the mobile phase at the flow rate of 1.0 m L/min;the column temperature was maintained at 30℃.The temperature of drift tube was 42℃ and the flow rare of N2 was 2.5 m L/min.Results The logarithm values of injection volumes exhibited good liner relationship with the logarithm value of peak areas when the injection volumes of saikosaponins a,b2,b1,and d were 0.0821-3.2080μg(r=0.9985),0.2673-10.4400μg(r=0.9998),0.08535-3.2600μg(r=0.9993),0.0909-3.5520μg(r=0.9997).The average recoveries were 101.61%,99.64%,95.36%,95.26%,respectively,and the RSDs were all less than 2%(n=6).Conclusion The method is simple in operation,high in accuracy and strong in specificity,and can be used for the determination of 4 saikosaponins in Huayu Quban Capsules.