液相色谱-质谱联用和酶增强免疫分析法测定人全血环孢素A浓度比较  

Comparison of Detection of Cyclosporin A in Human Blood by EMIT and LC-MS/MS Methods

作　　者：王海东[1] 倪善红[1] 孙增先[1] 

机构地区：[1]江苏省连云港市第一人民医院临床药学中心,连云港222002

出　　处：《医药导报》2016年第10期1064-1068,共5页Herald of Medicine

基　　金：连云港市第一人民医院青年英才豪森基金项目(QN130301)

摘　　要：目的建立液相色谱-质谱联用(LC-MS/MS)法测定人全血环孢素A(CsA)浓度,并与酶增强免疫分析(EMIT)法进行相关性分析。方法以乙腈为沉淀剂,环孢素D(CsD)为内标,Waters Atlantisd C18(3.9 mm×150 mm,3μm)色谱柱为分析柱,10 mmol·L^(-1)醋酸铵乙腈-10 mmol·L^(-1)醋酸铵为流动相进行梯度洗脱,质谱方法应用电喷雾电离源(ESI源),采用多反应监测方式进行正离子检测,用于定量分析的离子反应为CsA m/z 1 220.0→m/z 1 202.8,内标CsD m/z1 216.8→m/z 119.4。分别采用LC-MS/MS法和EMIT法测定124例患者服用CsA后的全血药物浓度,比较两种方法测定结果的相关性。结果以LC-MS/MS法测定结果(X)与EMIT法测定结果(Y)作线性回归方程,Y=0.999 3X+20.387(r=0.983,n=124,P<0.05),EMIT法测全血CsA的浓度较LC-MS/MS法偏高。结论 LC-MS/MS法和EMIT法测定人全血CsA的浓度相关性良好,在CsA药物浓度监测中,对不同测定方法的测定差异,应予以关注并作相应调整。Objective To establish an LC-MS/MS method for determination of cyclosporin A （CsA） concentrations in human blood,and to evaluate its relationship to concentrations determined by enzyme multiplied immunoassay technique （EMIT）method. Methods Blood sample was protein precipitated with acetonitrile, cyclosporin D （CsD）as the internal standard （IS）.Waters Atlantisd C18 column （3.9 mm×150 mm,3 μm）, 10 mmol·L-1 ammonium acetateacetonitrile-10 mmol·L-1 ammonium acetateas was the mobile phase by gradient elution.An tandem mass spectrometer was operated using an electrosprasy source （ESI） in the positive ion mode,operating in the multiple reaction monitoring （MRM） of the transitions of m/z 1 220.0？ m/z1 202.8 （CsA）,m/z 1 216.8？ m/z119.4 （CsD,IS）.Both EMIT and LC-MS/MS were applied to determine the concentration of CsA in blood of 124 volunteers.The results were analyzed to obtain correlation. Results The regression equation of resuits determined by LC-MS/MS （X） and EMIT （Y） was Y=0.999 3X＋20.387 （r=0.983,n=124,P〈0.05）.The concentration of CsA determined by EMIT was higher than that by LC-MS/MS. Conclusion The results determined by LC-MS/MS and EMIT methods have high correlation.The two detection systems possibly have system error.Difference of CsA monitoring by EMIT and LC-MS/MS during routine clinical pharmacotherapy should pay attention and make some adjustment.

关 键 词：环孢素A 环孢素D 酶增强免疫分析法 液相色谱-质谱联用法 

分 类 号：R979.5[医药卫生—药品]