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目的建立液相色谱-质谱联用(LC-MS)法测定人血清中槲皮素浓度。方法人血清样品经丙酮-甲醇提取,β-葡萄糖醛酸酶水解;采用色谱柱Agilent Poroshell 120 EC C_(18)(50 mm×4.6 mm,2.7μm),以乙腈(B)和1%甲酸水溶液(A)为流动相,梯度洗脱;流速1.0 m L/min,柱温40℃;电喷雾离子源,单四级杆质谱检测器,选择性负离子监测模式检测(ESI~-),检测离子质荷比为301.0(槲皮素),285.1(漆黄素);测定人血清中槲皮素浓度。结果槲皮素在1~200 ng/m L范围内线性关系良好,相关系数为0.9998;检出限为0.025 ng/m L;日内RSD为4.98%~8.35%,日间RSD为7.62%~9.73%;回收率为97.4%~112.8%(n=8)。结论 LC-MS法灵敏度较高、重复性良好,操作简单、快速,可用于人血清中槲皮素浓度的分析测定。
Objective To establish a method for the determination of quercetin in human serum by liquid chromatography-mass spectrometry (LC-MS). Methods The human serum samples were extracted with acetone-methanol and hydrolyzed by β-glucuronidase. The column was eluted with acetonitrile (B) and 1% formic acid aqueous solution using Agilent Poroshell 120 EC C 18 (50 mm × 4.6 mm, 2.7 μm) (A) as the mobile phase, gradient elution; flow rate 1.0 m L / min, column temperature 40 ℃; electrospray ionization source, single quadrupole mass spectrometry detector, selective negative ion monitoring mode detection The mass-to-charge ratio was 301.0 (quercetin), 285.1 (emodin); the concentration of quercetin in human serum was determined. Results The linearity of quercetin in the range of 1 ~ 200 ng / m L was good with a correlation coefficient of 0.9998. The detection limit was 0.025 ng / m L. The intraday RSD was 4.98% ~ 8.35% and the intraday RSD was 7.62% ~ 9.73 %; Recovery was 97.4% ~ 112.8% (n = 8). Conclusion LC-MS method has high sensitivity, good repeatability, simple and rapid operation, and can be used for the determination of quercetin in human serum.