槲皮苷、槲皮素及其甲基化产物的高压液相-质谱测定方法

THE SIMULTANEOUS DETERMINATION OF QUERCITRIN,QUERCETIN AND ITS METHYLATED METABOLITES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY

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 develop a high performance liquid chromatography-mass spectrometry（HPLC-MS） method for determination of quercitrin,quercetin,isorhamnetin and tamarixetin.Methods With buspirone as internal standard,quercitrin,quercetin,isorhamnetin and tamarixetin were determined simultaneously by HPLC-MS.The samples were separated by an Agilent-C18 and analyzed in positive ion electrospray mode.The mobile phase consisted of methanol and acetonitrile（1：1）,containing 0.1% formic acid.Results The calibration curve for quercetin showed good linearity in the range of 2-2000 ng/ml and others in 1-1000 ng/ml,with correlation coefficients of 0.99998,0.99712,0.99862,0.99806 respectively.Both accuracy and precision were satisfactory.Conclusion The method is simple,sensitive,precise and suitable for rapid determination of quercitrin,quercetin,isorhamnetin and tamarixetin simultaneously in cell culture samples.