目的：建立人全血中他克莫司浓度的LC-MS/MS 测定法,并用于他克莫司胶囊受试或参比药物在中国健康男性中单剂量空腹给药的一致性预评估。方法：采用单剂量给药,四周期、双序列、完全重复交叉、参比药物校正的平均生物等效性实验（RSABE）设计,12 名健康受试者单剂量交叉口服1 mg 他克莫司胶囊受试或参比药物后,0~120 h 内间隔取血,对全血提取富集后测定他克莫司血药浓度,用WinNonlin7.0 软件计算药动学参数并进行一致性预评估。结果：2 次服用参比药物后主要药动学参数Cmax、AUC0-τ、AUC0-∞个体内标准差均>0.294,用RSABE的判断标准,参比药物与受试药物主要药代动力学参数Cmax、AUC0-τ、AUC0-∞的单侧95% 置信区间上限（critbound）≤ 0,几何均数比值（GMR）点估计值（pointest）不超过0.80~1.25。故可以判定他克莫司胶囊受试与参比药物具有生物等效应。结论：建立的LC-MS/MS 法准确灵敏度高,可用于他克莫司胶囊的药代动力学研究和临床一致性的评价。
Objective: To establish the LC-MS/MS method for the determination of tacrolimus in human whole blood and to test the clinical consistency of tacrolimus test and reference capsules in healthy male Chinese subjects after single oral administration. Methods: Single-dose,4-way,fully replicated crossover design was adopted and the main pharmacokinetic parameters of tacrolimus were analyzed by RSABE. 12 healthy male volunteers were administered respectively with a single oral dose of test or reference capsules containing 1 mg of tacrolimus. The whole blood samples were collected at predetermined time intervals up to 120 h. The concentration of tacrolimus was determined after extraction and enrichment of whole blood. The pharmacokinetic statistical analysis was performed by Phoenix WinNonlin 7.0. Results: The results showed that the within-subject standard deviation of Cmax,AUC0-τ and AUC0-∞ were all greater than 0.294 when the volunteers were taken the reference drug twice. According to RSABE criteria the upper bound of 95% confidence interval（critbound） of Cmax,AUC0-τ and AUC0-∞ of reference and test preparations were all less than 0. And the geometric mean ratio（pointest） of Cmax,AUC0-τ and AUC0-∞ of reference and test preparations were all within the range of 0.80 and 1.25. The results demonstrated that the test and reference capsules were clinically consistent. Conclusion: The LC-MS/MS method has been validated to be accurate and sensitive. The method can be used for pharmacokinetic studies and clinical consistency evaluation of tacrolimus capsules.
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