用HPLC-MS/MS法测定儿童微量血浆中地高辛浓度  

HPLC-MS/MS method for monitoring digoxin in children with small volume of plasma

作　　者：杨志梅 冯章英[2] 秦亚彬 YANG Zhi-mei;FENG Zhang-ying;QIN Ya-bin(Department of Emergency,the Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei Province,China;Department of Clinical Pharmacology,the Fourth Affiliated Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei Province,China;Department of Pharmacy,Children's Hospital of Hebei Province,Shijiazhuang 050031,Hebei Province,China)

机构地区：[1]河北医科大学第四医院急诊科,河北石家庄050011 [2]河北医科大学第四医院临床药理研究部,河北石家庄050011 [3]河北省儿童医院药学部,河北石家庄050031

出　　处：《中国临床药理学杂志》2024年第9期1331-1335,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河北省医学科学研究课题计划项目(20221335);河北省卫生健康创新专项基金资助项目(22377729D)。

摘　　要：目的建立液相色谱-串联质谱(HPLC-MS/MS)法检测血浆中地高辛含量,并用于患儿治疗药物监测(TDM)。方法取血浆样本10μL经乙腈沉淀蛋白,用Phenomenex Kinetex EVO C18(30.0 mm×2.1 mm,2.6μm)色谱柱分离,流动相为水(含0.05%甲酸和2 mmol·L^(-1)乙酸铵)和甲醇,梯度洗脱,流速:0.55 mL·min^(-1),进样体积:30μL,柱温:45℃。电喷雾离子源正离子模式,动态多反应监测(dMRM)扫描分析,离子通道分别选择为m/z 798.3→651.3(地高辛)和m/z 801.3→654.3(地高辛-d3)。考察该方法的专属性、标准曲线与定量下限、准确度和精密度、提取回收率、基质效应以及稳定性。结果地高辛及内标的保留时间均为1.39 min,地高辛在0.25~8.00 ng·mL^(-1)线性良好,代表性的标准曲线方程为y=18.08×10^(-2)x+29.71×10^(-4)(r^(2)=0.99577),批内和批间精密度相对标准偏差均小于15%,提取回收率为91.00%~99.50%,稳定性良好,基质效应符合规定。本方法成功用于65例患儿的地高辛血药浓度测定。结论本方法操作简便、快速、准确、专属性强,可用于患儿临床治疗药物监测。Objective To establish a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for therapeutic drug monitoring(TDM)of digoxin in children.Methods The plasma samples 10μL were precipitated protein by acetonitrile.Then,the analytes were gradient eluted on a Phenomenex Kinetex EVO C18 column(30.0 mm×2.1 mm,2.6μm)by HPLC-MS/MS with mobile phase consisted of water(0.05%formic acid and 2 mmol·L^(-1)ammonium acetate)-methanol at the flow rate of 0.55 mL·min^(-1).The mass detection was performed in positive ionization mode using the electrospray ionization(ESI)source.Dynamic multiple reaction monitoring mode(dMRM)was used to quantify at m/z 798.6→651.5(digoxin)and m/z 801.6→654.5(digoxin-d3),respectively.The selectivity,standard curve,precision and extraction recoveries,matrix effect,and stability were investigated.Results The retention time of both digoxin and internal standard was 1.39 min.The linear range of digoxin in human plasma was among 0.25-8.00 ng·mL^(-1),a typical standard curve was y=18.08×10^(-2)x+29.71×10^(-4)(r^(2)=0.99577).The intra-day and inter-day accuracies of digoxin were less than 15%.Extraction recovery rate was 91.00%-99.50%.Stability studies showed good results.Matrix factor normalized by internal standard was within acceptance criteria.The measurement results were in line with the national requirement of External quality evaluation.This method was fully verified and applied to determine the plasma digoxin concentrations of 65 pediatric patients.Conclusion This LC-MS/MS method was simple,rapid and accurate.It is approved appropriate and practical for the therapeutic drug monitoring of digoxin in routine clinical laboratory practice,especially for children.

关 键 词：地高辛 液相色谱-串联质谱法 儿童 治疗药物监测 蛋白沉淀 

分 类 号：R97[医药卫生—药品]