高效液相色谱-串联质谱法测定儿童微量脑脊液中美罗培南浓度

Determination of Meropenem concentration in children’s trace cerebrospinal fluid by high performance liquid chromatography-tandem mass spectrometry

目的建立高效液相色谱-串联质谱法(HPLC-MS/MS)快速测定儿童微量脑脊液中美罗培南的浓度。方法以厄他培南为内标,10μl脑脊液样品经0.1%甲酸-乙腈沉淀蛋白,液相分离采用Hypersil GOLD^(TM)色谱柱(50 mm×2.1 mm,3μm),以0.1%甲酸-水和0.1%甲酸-乙腈为流动相,梯度洗脱,流速0.5 ml/min,柱温40℃;采用电喷雾离子源,正离子多反应监测模式检测。对2020年2月至3月河北省儿童医院神经内科收集的样品进行分析。结果脑脊液中美罗培南浓度在0.01~50.00μg/ml内线性良好,线性方程为Y=0.880X+0.000168(r=0.9989),定量下限为0.01μg/ml;提取回收率为(93.24±9.48)%~(104.62±4.63)%;美罗培南在脑脊液中无明显基质效应。患儿脑脊液样品中美罗培南浓度在0.10~1.25μg/ml。结论本法样品量少、简单、快速、准确、灵敏,适用于儿童微量脑脊液中美罗培南的浓度监测。

Objective To establish a rapid high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for the rapid determination of Meropenem in children’s trace cerebrospinal fluid.Methods Using Ertapenem as the internal standard,10μl cerebrospinal fluid samples were precipitated by 0.1%formic acid-acetonitrile.The liquid phase separation was performed on a Hypersil GOLD^(TM) column(50 mm×2.1 mm,3μm)with 0.1%formic-water and 0.1%formic-acetonitrile as mobile phases,and gradient elution at the flow rate of 0.5 ml/min,and column temperature was 40℃;the detection was carried out by positive electrospray ionization mass spectrometry in multiple reaction monitoring mode.Samples collected from the Department of Neurology of Hebei Children’s Hospital from February 2020 to March 2020 were analyzed.Results The concentration of Meropenem in cerebrospinal fluid was in the range of 0.01-50.00μg/ml,and the linear equation was Y=0.880X+0.000168(r=0.9989),the lowest detection limit was 0.01μg/ml;the extraction recoveries were(93.24±9.48)%-(104.62±4.63)%;Meropenem had no significant matrix effect in cerebrospinal fluid.The concentration of Meropenem in cerebrospinal fluid samples was 0.10-1.25μg/ml.Conclusion The method is sample-saving,simple,rapid,accurate,and sensitive,and is suitable for the concentration monitoring of Meropenem in children’s trace cerebrospinal fluid.