LC-MS/MS法测定人血浆中地高辛的浓度及其在儿童患者中的应用

Determination of digoxin in human plasma by LC-MS/MS and its application in pediatric patients

建立了简单、快速、可靠的LC-MS/MS法测定人血浆中地高辛的浓度。以地高辛-d3为内标,血浆样品经液液萃取法处理后进样。色谱柱为Kinetex C_(18)(2.1 mm×50 mm,2.6μm;Phenomenex),流动相为均含1 mmol/L醋酸铵和1 mmol/L甲酸的水溶液和乙腈溶液(55∶45),采用等度洗脱,样品运行时间仅为2 min。选择TripleQuad^(TM)4500MD质谱仪,在电喷雾离子化方式下(ESI),以正离子模式进行多重反应监测(MRM),地高辛及其内标的监测离子对分别为m/z 798.5→651.3和m/z 801.6→654.4。结果表明,在0.100~20.0 ng/mL范围内,标准曲线线性关系良好,该方法选择性好,准确度、精密度符合要求,各浓度水平提取回收率平行,稳定性良好,无基质效应等影响。该方法成功用于接受地高辛治疗的患儿的血样检测,可更大程度增加疗效、避免毒性反应,为临床个体化用药提供指导和依据。

The aim of the study was to develop a simple,rapid and accurate LC-MS/MS method for the determi⁃nation of digoxin.Digoxin-d3 was taken as the internal standard(IS),and sample preparation was achieved by liquid-liquid extraction.Chromatographic separation was performed on a Kinetex C_(18) column(2.1 mm×50 mm,2.6μm;Phenomenex)using an isocratic elution with merely 2 min for each sample.The mobile phase consist⁃ed of water and acetonitrile solutions,both containing 1 mmol/L ammonium acetate and 1 mmol/L formic acid(55∶45).The detection was conducted on a TripleQuad^(TM) 4500MD mass spectrometer coupled with electrospray ionization interface under positive-ion multiple reaction monitoring mode.The transitions were m/z 798.5→651.3 and m/z 801.6→654.4 for digoxin and digoxin-d3,respectively.Results showed that the method was linear over the range of 0.100-20.0 ng/mL.The selectivity,accuracy and precision,recovery and stability of the method were all within the acceptable limits with no matrix effect.This method was successfully applied to a girl treated with digoxin with substantial improvement of therapeutic effect and elimination of toxic reaction,so it can provide valuable fuidance and reference for individualized medication in clinical practice.