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目的建立同时测定槲皮苷、槲皮素、异鼠李亭、柽柳黄素的高效液相色谱-质谱(HPLC-MS)测定方法。方法以丁螺环酮为内标,将槲皮苷、槲皮素、异鼠李亭和柽柳黄素溶解于磷酸缓冲液(PBS)中,进行HPLC-MS测定。色谱柱为Agilent-C18柱,流动相为甲醇:乙腈(1:1)(含0.1%甲酸),等梯度洗脱,离子源为ESI(+),采用选择离子监测(SIM)方式检测。结果槲皮素在2～2000ng/ml范围内线性良好,槲皮苷、异鼠李亭、柽柳黄素在1～1000 ng/ml范围内线性良好,相关系数分别为0.99998、0.99712、0.99862、0.99806;方法的准确性和精密度良好。结论该方法操作简单、灵敏度高、准确性好,可应用于细胞培养样品中槲皮苷、槲皮素及其甲基化代谢产物的测定。
Objective To establish a method for the simultaneous determination of quercetin, quercetin, isorhammitin, and otinexanthin by high performance liquid chromatography-mass spectrometry (HPLC-MS). Methods Buspirone was used as an internal standard, and quercetin, quercetin, isorhamipitine and 柽liquidin were dissolved in phosphate buffered saline (PBS), and the contents of quercetin, quercetin, and isotin were determined by HPLC-MS. The column was Agilent-C18. The mobile phase consisted of methanol: acetonitrile (1: 1) with 0.1% formic acid and the isocratic gradient. The ion source was ESI (+) and detected by SIM. Results The calibration curve of quercetin was linear in the range of 2 ~ 2000ng / ml. The linearities of quercetin, isorhamipitine and 柽liuutin in the range of 1 ~ 1000 ng / ml were good with the correlation coefficients of 0.99998, 0.99712, 0.99862 and 0.99806, respectively The accuracy and precision of the method are good. Conclusion The method is simple, sensitive, accurate and can be applied to the determination of quercetin, quercetin and their methylated metabolites in cell culture samples.