在中国心脏移植患者中高效液相色谱串联质谱法和化学发光微粒子免疫分析法测定他克莫司血药浓度的比较

Quality consistency evaluation for determination of whole-blood tacrolimus concentration in heart-transplant recipients by two methods: High performance liquid chromatography-tandem mass spectrometry method and chemiluminescence microparticle immuno assay

目的比较高效液相色谱-串联质谱法(HPLC-MS/MS)和化学发光微粒子免疫检测技术(CMIA)测定人全血他克莫司(FK506)浓度结果的相关性,以及2种方法在他克莫司浓度监测工作中的应用。方法收集253例心脏移植患者(n=581)术后服用他克莫司的全血样本,采用HPLC-MS/MS法测定人全血他克莫司血药浓度,通过专属性、标准曲线与定量下限、精密度、准确度、基质效应和提取回收率以及稳定性试验进行方法学验证;并与CMIA法的测定结果进行比较,观察、比较2种检测手段的测定结果及相关性。结果用HPLC-MS/MS测定人全血他克莫司浓度,线性范围2~30 ng·mL^(-1),定量下限为2ng·mL^(-1);日内及日间精密度(RSD%)均小于15%。按照Bland-Altman法计算,95.52%Lo A=(0.22,5.62),95.52%的差值都位于一致限内,两种方法测定值一致性良好。回归方程为FK506_(HPLC-MS/MS)=0.70×FK506_(CMIA)-0.17,Pearson相关系数为0.902 4(P<0.05),说明两种方法具有良好的相关性。结论HPLC-MS/MS法测定他克莫司血药浓度定量特异性强,灵敏度高。与CMIA法相比,HPLC-MS/MS测定值低,对他克莫司母药有高度的选择性。

Objective To analyze the correlation between high performance liquid chromatography-tandem mass spectrometry（ HPLC-MS/MS） method and chemiluminescence microparticle immuno assay（ CMIA） for determination of tacrolimus concentration in human blood,and explore the application of the two methods in clinical therapeutic drug monitoring. Methods A total of 253 heart transplant patients（ n = 581）were enrolled. The concentration of tacrolimus in whole blood was determined by HPLC-MS/MS. The specificity,standard curve and quantification limit,precision,accuracy,matrix effect and extraction recovery rate and stability test was performed. CMIA was used simultaneously,the results of two methods were observed and compared. Results The linear range by HPLC-MS/MS was 2-30 ng·mL^（-1）,and the lower limit of quantification was 2 ng · mL^（-1）. The intra-and inter-day precision（ RSD%） by HPLC-MS/MS was less than 15%. According to theBland-Altman method,the percentage of the limit can be obtained： 95. 52% Lo A =（ 0. 22,5. 62）. And 95. 52%difference value was within the limit and the consistency of the two methods was good. According to the Pearson linear regression analysis,the regression equation was FK506_（HPLC-MS/MS）= 0. 70 × FK506_（CMIA）-0. 17 and correlation coefficient was 0. 902 4（ P 0. 05） with good correlation. Conclusion The method of HPLC-MS/MS for the determination of tacrolimus blood concentration shows a high specificity and sensitivity. Compared with the results of CMIA,the measured value by HPLC-MS/MS is low,but HPLC-MS/MS is highly selective for parent drug of tacrolimus.