ELBLAB GmbH Zentrum für Labor Medizin Meißen Riesa Radebeul, Weinbergstraße 8, 01589 Riesa, Germany.
Department of Analytical Chemistry, Institute of Chemistry, Faculty of Science, Pavol Jozef Šafárik University in Košice, Moyzesova 11, SK-041 54 Košice, Slovak Republic.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Oct 1;1128:121772. doi: 10.1016/j.jchromb.2019.121772. Epub 2019 Aug 20.
A rapid, simple and robust HPLC-MS/MS method for simultaneous determination of immunosuppressants Cyclosporine A, Tacrolimus, Sirolimus and Everolimus has been developed and validated. Sample of whole blood with volume of 50 μL was prepared by a protein precipitation with methanol and 0.5 mol. L ZnSO. Chromatographic separation was achieved on a Phenyl-Hexyl column by a gradient elution using 20 mmol.L ammonium formate/0.1% (v/v) formic acid in water (mobile phase A) and 20 mmol.L ammonium formate/0.1% (v/v) formic acid in methanol (mobile phase B) with flow rate 1 mL.min. The run time was 3.5 min. Electrospray ionization and multiple reaction monitoring was used. The lower limit of quantification (LLOQ) was set at 0.5 μg.L for Tacrolimus, Sirolimus and Everolimus and 5 μg.L for Cyclosporine A. The method demonstrated adequate accuracy and precision with sufficient linearity range.
建立并验证了一种快速、简单、稳健的 HPLC-MS/MS 法，用于同时测定免疫抑制剂环孢素 A、他克莫司、西罗莫司和依维莫司。用甲醇和 0.5mol·L ZnSO4 沉淀全血 50μL 作为样品。采用苯基-己基柱，以 20mmol·L 甲酸铵/0.1%（v/v）甲酸水溶液（流动相 A）和 20mmol·L 甲酸铵/0.1%（v/v）甲醇（流动相 B）梯度洗脱，流速为 1mL·min。运行时间为 3.5min。采用电喷雾电离和多反应监测模式。他克莫司、西罗莫司和依维莫司的定量下限（LLOQ）设定为 0.5μg·L，环孢素 A 的LLOQ 设定为 5μg·L。该方法具有足够的线性范围，准确度和精密度良好。
