摘要
目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)法同时测定芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚在大鼠血浆中浓度的方法.方法采用AB SciexQtrap5500三重四极杆液质联用系统,色谱柱为Waters UPLC BEH C18柱(100.0 mm× 2.1mm,1.7 μm),体积流量为0.3 mL/min,柱温为25.0 ℃,进样量为3 μL,流动相为0.1％甲酸-乙腈梯度洗脱.采用电喷雾离子源,多反应监测模式进行检测.结果:芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚分别在 1.000～64.000、1.000～1 000.000、0.300～32.000、1.000～250.000和 1.000～64.000 µg/mL质量浓度范围内与峰面积呈良好的线性关系,标准曲线的相关系数均在0.99～1.00之间,大黄素的定量限为0.3 μg/mL,其余4种大黄蒽醌成分定量限均为1.0µg/mL;精密度和稳定性试验的RSD值均小于8.7％;平均加样回收率为88.7％～98.8％,RSD值为0.9％～2.4％;基质效应在89.8％～96.2％之间.结论:本研究建立的UPLC-MS/MS法简单、快速、准确灵敏,可用于大黄蒽醌苷元在大鼠体内的含量测定.
Abstract
Objective:To establish a UPLC-MS/MS method for the simultaneous measurement of rat plasma concentrations of aloe-emodin,rhein,emodin,chrysophanol and physcion.Methods:The AB SciexQtrap5500 triple quadrupole LC/MS system was used for the Waters UPLC BEH C18 column(100.0 mm×2.1 mm,1.7 μm),with a volume flow rate of 0.3 mUmin,a column temperature of 25.0 ℃,an injection volume of 3 µL,and a mobile phase of 0.1％formic acid-acetonitrile gradient elute.The electron spray ionization was adopted,and the working mode was a multi-reaction monitoring mode.Results:Aloe-emodin,rhein,emodin,chrysophanol and physcion showed good linear relationships with peak areas in the concentration ranges of 1.000-64.000,1.000-1 000.000,0.300-32.000,1.000-250.000 and 1.000-64.000 μg/mL,respectively,with the correlation coefficients of the standard curves in the range of 0.99-1.00.The limits of quantification(LOQs)of emodin were 0.3 μg/mL,and those of the other four rhubarb anthraquinones were 1.0 μg/mL.The RSD of the precision and stability tests were less than 8.7％.The average recoveries of spiked standards ranged from 88.7％to 98.8％,with RSD of 0.9％-2.4％,and the matrix effects ranged from 89.8％to 96.2％.Conclusion:The UPLC-MS/MS approach developed in this study is straightforward,quick,precise,and sensitive,and it may be utilized to estimate the anthraquinonesa-pogenins concentration of Dahuang(rhubarb)in rats.
基金项目
河北省中央引导地方科技发展资金项目科技创新基地项目(216Z2502G)
河北省自然科学基金中医药联合基金(H2022406043)
承德医学院学科建设项目(2023LJ-1)
承德医学院高层次人才科研启动基金(202210)
维普
万方数据