HPLC-MS/MS法同时测定紫菀中9种化学成分

Simultaneous determination of nine constituents in Asteris Radix by HPLC-MS/MS

目的建立HPLC-MS/MS法同时测定紫菀中山柰酚、槲皮素、木犀草素、异鼠李素、东莨菪内酯、伞形花内酯、阿魏酸、咖啡酸和绿原酸9种化学成分的方法。方法采用Diamonsil C18柱（150 mm×4.6 mm,5μm）,流动相为水（0.05%甲酸）-乙腈（0.05%甲酸）,梯度洗脱,体积流量0.8 m L/min,进样量10μL,柱温30℃;采用电喷雾离子源进行负离子模式监测,多反应监测模式（MRM）用于定量分析,源喷射电压为-4 500 V,离子源温度为650℃。结果山柰酚、槲皮素、木犀草素、异鼠李素、东莨菪内酯、伞形花内酯、阿魏酸、咖啡酸和绿原酸分别在0.970～30.30μg/m L（r=0.998 7）、0.498～15.55μg/m L（r=0.999 6）、0.014～0.438μg/m L（r=0.999 5）、0.022～21.95μg/m L（r=0.998 5）、0.011～1.100μg/m L（r=0.999 5）、0.008～0.247μg/m L（r=0.999 8）、0.194～6.050μg/m L（r=0.999 6）、0.197～6.150μg/m L（r=0.999 5）和1.459～45.609μg/m L（r=0.999 0）线性关系良好;平均回收率在97.43%～103.87%,精密度RSD值为1.95%～3.09%。结论本法简便、快速、灵敏度高、专属性好,可用于紫菀中9种化学成分的同时测定。

Objective To develop an HPLC-MS/MS method for the simultaneous determination of nine constituents（kaempferol, quercetin, luteolin, isorhamnetin, scopoletin, umbelliferone, ferulic acid, caffeic acid, and chlorogenic acid） in Asteris Radix（the roots and rhizoma of Aster tataricus. Methods Analysis was performed on a Diamonsil C18 column（150mm × 4.6 mm, 5 μm） eluted with acetonitrile（0.05% formic acid） and water（0.05% formic acid） in a gradient program. The flow rate was 0.8 m L/min, the injection volume was 10 μ L, and the column temperature was 30 ℃. The multiple-reaction monitoring scanning（MRM） was employed for the quantification with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at-4 500 V and the turbo spray temperature was maintained at 650 ℃. Results Kaempferol, quercetin, luteolin, isorhamnetin, scopoletin, umbelliferone, ferulic acid, caffeic acid, and chlorogenic acid showed a good linearity in the ranges of 0.970—30.30（r = 0.998 7）, 0.498—15.55（r = 0.999 6）, 0.014—0.438（r = 0.999 5）, 0.022—21.95（r = 0.998 5）, 0.011—1.100（r = 0.999 5）, 0.008—0.247（r = 0.999 8）, 0.194—6.050（r = 0.999 6）, 0.197—6.150（r = 0.9995）, and 1.459 — 45.609 μ g/m L（r = 0.999 0）. The average recoveries varied from 97.43% — 103.87%. The RSD values of precision varied from 1.95%- 3.09%. Conclusion The method is simple, rapid, and sensitive. It could be used as a quantitative determination method for the nine components in A. tataricus.