UPLC-MS/MS同时测定菟丝子中6种黄酮类成分的含量

Simultaneous Determination of Six Flavonoids in Semen Cuscutae by UPLC-MS/MS

目的:建立同时测定菟丝子中6种黄酮类化合物成分含量的高效快速的超高效液相色谱-串联质谱法(UPLC-MS/MS)方法。方法:采用Waters UPLC BEH C18色谱柱(50 mm×2.1 mm,1.7μm),甲醇-0.1%甲酸水溶液为流动相,梯度洗脱,流速为0.3 ml/min,柱温为30℃;质谱条件采用ESI-离子源,多反应离子检测模式(multiple reaction monitoring,MRM),对菟丝子中6种黄酮类成分金丝桃苷、紫云英苷、异槲皮苷、槲皮苷、山萘酚、槲皮素进行含量测定。结果:在上述条件下测得各化合物分别在5.0~500 ng/ml线性范围内呈良好线性关系(r>0.999);精密度、重复性、回收率、稳定性均满足方法学验证要求。结论:该方法高效、准确、灵敏,可用于同时测定菟丝子中金丝桃苷、紫云英苷、异槲皮苷、槲皮苷、山萘酚、槲皮素的含量。

Objective:To develop a UPLC-MS/MS method for simultaneous determination of six flavonoids in Semen Cuscutae.Methods:The separation of the six flavonoids was performed on an ACQUITY UPLC BEH C18 column(50 mm×2.1 mm,1.7μm)at 30℃.The mobile phase consisted of methanol and aqueous 0.1%formic acid using gradient elution,which was delivered at a flow rate of 0.3 ml/min.The determination of the Hyperoside,Astragalin,Isoquercitrin,Quercitrin,Quercetin,and Kaempferol from Semen Cuscutae was performed on UPLC-MS/MS system with turbo ion spray source in the negative ion and multiple reactionmonitoring mode.Results:The method was linear over the concentration range of 5.0-500 ng/ml for the six analytes.All calibration curves showed good linearity(r>0.999).The precision,repeatability,recoveries were all conformed to the validated requirements of methodology.Conclusion:The developed and validated methods are efficient,accurate and selective for the separation and quantification of the six flavonoids in Semen Cuscutae.