超高效液相色谱串联质谱法测定护肝片(胶囊)中柴胡皂苷K含量

Content Determination of Nepasaikosaponin K in Hugan Tablets(Capsules)by UPLC-MS/MS

目的建立测定护肝片(胶囊)中柴胡皂苷K含量的超高效液相色谱串联质谱法。方法色谱柱为Waters Acquity UPLC BEH-C_(18)柱(50 mm×2.1 mm,1.7μm),流动相为乙腈-水(梯度洗脱),流速为0.30 mL/min,柱温为35℃,进样量为1μL。电喷雾离子源,负离子检测,多反应监测模式,离子源温度为150℃,毛细管电压为4.0 kV,脱溶剂温度为500℃,锥孔气流量(氮气)为50 L/h,碰撞气流速(氩气)为0.15 L/h,氮气流量为800 L/h。结果柴胡皂苷K质量浓度在0.2007~5.0176μg/mL范围内与峰面积线性关系良好(r=0.9999,n=5);检测限为0.05 mg/kg,定量限为0.20 mg/kg;精密度、稳定性、重复性试验结果的RSD均小于3.0%;平均加样回收率为98.43%,RSD为0.95%(n=9)。12个厂家47批样品中有28批的柴胡皂苷K含量介于0.5564~3.5899μg/g。结论该方法操作简便、快捷、灵敏度高、重复性好,可用于护肝片(胶囊)的质量控制。

Objective To establish an ultra-high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the determination of nepasaikosaponin K in Hugan Tablets(Capsules).Methods The chromatographic column was Waters Acquity UPLC BEH-C_(18) column(50 mm×2.1 mm,1.7μm),the mobile phase was acetonitrile-water(gradient elution),the flow rate was 0.30 mL/min,the column temperature was 35℃,and the injection volume was 1μL.The electrospray ion source(ESI)was adopted with negative ion detection and multiple reaction monitoring(MRM)mode,the ion source temperature was 150℃,the capillary voltage was 4.0 kV,the desolvent temperature was 500℃,the cone hole gas flow(nitrogen)was 50 L/h,the collision gas flow(argon)was 0.15 L/h,and the nitrogen flow was 800 L/h.Results The linear range of nepasaikosaponin K was 0.2007-5.0176μg/mL(r=0.9999,n=5).The limit of detection was 0.05 mg/kg,and the limit of quantitation was 0.20 mg/kg.The RSDs of precision,stability and repeatability tests were all lower than 3.0%.The average recovery rate of nepasaikosaponin K was 98.43%with an RSD of 0.95%(n=9).Among the 47 batches of samples from 12 manufacturers,the content of nepasaikosaponin K in 28 batches of samples was in the range of 0.5564 to 3.5899μg/g.Conclusion This method is simple,fast,sensitive and repeatable,which can be used for the quality control of Hugan Tablets(Capsules).