液相色谱串联质谱法测定人血浆中硫酸黏菌素的浓度

Determination of colistin sulfate in human plasma by LC-MS/MS

目的建立液相色谱串联质谱(LC-MS/MS)法测定人血浆中硫酸黏菌素的浓度。方法内标选用多黏菌素B,血浆样品采用甲醇-5%三氯乙酸(50∶50,V/V)沉淀蛋白。色谱柱为Dikma C_(18)(4.6 mm×150 mm,5μm),流动相A为水(含0.1%甲酸),流动相B为乙腈(含0.1%甲酸),梯度洗脱,流速为0.8 mL·min^(-1),柱温为40℃,进样量为10μL。采用电喷雾离子化,正离子模式下进行多反应监测多黏菌素E_(1)(m/z 585.7→101.2)、多黏菌素E_(2)(m/z 578.8→101.2)和内标多黏菌素B(m/z 602.7→241.4)。结果硫酸黏菌素在0.1~10μg·mL^(-1)内与峰面积线性关系良好(r=0.9982),定量限为0.1μg·mL^(-1),日内与日间精密度、准确度、提取回收率、基质效应、稳定性均符合方法学要求。结论本方法简便、快速、灵敏,可用于测定人体内硫酸黏菌素的浓度。

Objective To determine colistin sulfate in human plasma by LC-MS/MS.Methods The plasma samples were treated by the protein precipitation method with methanol-5%trichloroacetic acid solution(50∶50,V/V)and the internal standard was polymyxin B.The Dikma C_(18)(4.6 mm×150 mm,5μm)was used.Gradient elution was conducted with mobile phase A(aqueous solution containing 0.1%formic acid)and mobile phase B(acetonitrile containing 0.1%formic acid).MS detection was used with electrospray ionization source operated in a multi-reaction monitoring mode.The ion transitions were m/z 585.7→101.2(polymyxin E_(1)),m/z 578.8→101.2(polymyxin E_(2)),and m/z 602.7→241.4(polymyxin B).Results The linearity of colistin sulfate was 0.1~10μg·mL^(-1)(r=0.9982).The intra-day and inter-day precision,accuracy,extraction recovery rate,matrix,and stability of colistin sulfate all met the methodological requirement.Conclusion The method is rapid,simple,and sensitive,and can be used to detect the concentration of colistin sulfate in human plasma.