超高效液相色谱-高分辨飞行时间质谱法测定人体血浆中雷莫拉宁的残留

Determination of ramoplanin residual in human plasma by ultra-high performance liquid chromatography couple with high resolution time of flight mass spectrometry

目的建立一种人体血浆中雷莫拉宁的超高效液相色谱串联高分辨飞行时间质谱(UHPLC-QTOF-MS)的检测新方法。方法 100μl血浆样品中加入300μl甲醇沉淀蛋白,4℃下反应30 min,离心,取上清液,氮吹至近干,再复溶于100μl 0.2%甲酸水溶液。采用超高效液相色谱串联高分辨飞行时间质谱,以BEH-C_(18)色谱柱分离,甲酸溶液(0.2%)-甲醇为流动相进行梯度洗脱,流速:0.30 ml/min,ESI+模式下进行质谱测试,外标法定量分析。结果雷莫拉宁的浓度在30μg/L~800μg/L时与其峰面积呈良好的线性关系(r=0.999 3),方法的检出限为10μg/L,定量下限为30μg/L,血浆空白样品3个质量浓度水平的加标回收率为84.4%~98.9%,3次测定结果的相对标准偏差(RSD)为0.8%~6.3%。结论该分析方法具有前处理简便、分析快速、灵敏准确的特点,对于雷莫拉宁在临床药物监测及在环境和食品中的残留评价具有重要意义。

Objective To establish a new analytical method for the determination of ramoplanin residual in human plasma by using ultra-high performance liquid chromatography couple with quadrupole time of flight mass spectrometry( UHPLC-QTOF-MS).Methods Typically, 100 μl of plasma sample was precipitated with 300 μl of methanol for 30 min at 4 ℃ and then separated by high speed centrifugation. The supernatant was evaporated by nitrogen to dryness and re-dissolved in 100 μl of 0. 2% formic acid solution. Separation of ramoplanin was achieved by a BEH-C_(18) column with mobile phases as methanol and 0. 2% formic solution at a flow rate of 0. 30 ml/min. Ramoplanin was detected by ultra-high performance liquid chromatography couple with high resolution time of flight mass spectrometry under ESI+mode and quantified by external standard method.Results Good linearity was obtained in the range of 30 μg/L-800 μg/L with the correlation coefficients(r) of 0. 999 3.The detection limit of ramoplanin was 10 μg/L and the quantitation limit was 30 μg/L. The spiked recoveries were in the range of 84. 4%-98. 9%, and the relative standard deviations(RSD) were 0. 8%-6. 3%(n= 3).Conclusion The results demonstrated that this method was simple, fast and sensitive, and was of great significance in clinical drug monitoring and the evaluation of ramoplanin residue in environment and food.