Paniagua-González Lucía, Lendoiro Elena, Otero-Antón Esteban, Molina-Pérez Esther, Varo-Pérez Evaristo, López-Rivadulla Manuel, Cruz Angelines, de-Castro-Ríos Ana
Servizo de Toxicoloxía, Instituto de Ciencias Forenses, Universidade de Santiago de Compostela, Rúa San Francisco, s/n, 15782 Santiago de Compostela, Spain.
Unidade de Transplante Abdominal, Complexo Hospitalario Universitario de Santiago de Compostela, Rúa da Choupana, s/n, 15706 Santiago de Compostela, Spain.
Bioanalysis. 2019 Aug;11(16):1509-1521. doi: 10.4155/bio-2019-0143. Epub 2019 Sep 11.
This study aimed: to develop and validate an LC-MS/MS method for mycophenolic acid, tacrolimus, sirolimus, everolimus and cyclosporin A in oral fluid (OF), as an essential tool to study the usefulness of OF as an alternative matrix for immunossuppressants' therapeutic drug monitoring; and to find the best OF collector for these analytes. Chromatographic separation was achieved using an XBridge Shield RP18 analytical column maintained at 65ºC, using 2 mM ammonium formate and 0.1% formic acid in water (A) and acetonitrile (B) as mobile phase. OF sample was extracted with solid phase extraction after sonication and protein precipitation. Method validation met all the acceptance criteria. LODs were 0.05-1 ng/ml, and LOQs 0.1-5 ng/ml. Silanized tubes offered the best recoveries. The method was successfully applied to 31 OF specimens, describing everolimus detection in OF for the first time. The proposed method is sensitive enough for the detection of OF trough concentrations in patients receiving immunosuppressants when using an appropriate OF collector.
开发并验证一种用于测定口腔液（OF）中霉酚酸、他克莫司、西罗莫司、依维莫司和环孢素A的液相色谱-串联质谱（LC-MS/MS）方法，作为研究OF作为免疫抑制剂治疗药物监测替代基质的有用性的重要工具；并找到针对这些分析物的最佳OF收集器。使用保持在65ºC的XBridge Shield RP18分析柱进行色谱分离，以2 mM甲酸铵和0.1%甲酸的水溶液（A）和乙腈（B）作为流动相。OF样品经超声处理和蛋白沉淀后用固相萃取法进行提取。方法验证符合所有验收标准。检测限为0.05 - 1 ng/ml，定量限为0.1 - 5 ng/ml。硅烷化管的回收率最佳。该方法成功应用于31份OF标本，首次描述了依维莫司在OF中的检测情况。当使用合适的OF收集器时，所提出的方法对于检测接受免疫抑制剂治疗的患者的OF谷浓度足够灵敏。
