未经证实:免疫抑制剂的治疗药物监测(TDM)对于移植患者的最佳护理至关重要。免疫分析和液相色谱-质谱(LC-MS)是最常用的TDM方法。然而,免疫测定可能受到来自异源性抗体和结构相似的药物和代谢物的干扰。此外,标称质量LC-MS测定可能难以优化,并且在可检测化合物的数量上受到限制。
UNASSIGNED:这项研究的目的是使用在线固相萃取(SPE)和精确质量全扫描单离子监测(FS-SIM)数据采集模式对免疫抑制剂TDM进行基于质谱的测试。
UNASSIGNED:LC-MS分析在具有Q-ExactivePlus质谱仪的TLX-2多通道HPLC上进行。TurboFlow在线SPE用于样品清理。将精确质量MS设置为具有FS-SIM的正电喷雾电离模式,以定量他克莫司,西罗莫司,依维莫司,和环孢菌素A。MS2片段模式用于化合物确认。
未经评估:该方法在精度方面进行了验证,分析偏差,定量极限,线性度结转,样品稳定性,和干扰。他克莫司的定量,西罗莫司,依维莫司,和环孢菌素A与独立参考实验室的结果密切相关(r=0.926-0.984)。
UNASSIGNED:准确质量FS-SIM可成功用于免疫抑制剂TDM,与标准方法产生的结果具有良好的相关性。TurboFlow在线SPE允许简单的“蛋白质崩溃和射击”样品制备方案。与传统MRM相比,通过FS-SIM的分析物定量促进了流线型的测定优化过程。
UNASSIGNED: Therapeutic drug monitoring (TDM) of immunosuppressants is essential for optimal care of transplant patients. Immunoassays and liquid chromatography-mass spectrometry (LC-MS) are the most commonly used methods for TDM. However, immunoassays can suffer from interference from heterophile antibodies and structurally similar drugs and metabolites. Additionally, nominal-mass LC-MS assays can be difficult to optimize and are limited in the number of detectable compounds.
UNASSIGNED: The aim of this study was to implement a mass spectrometry-based test for immunosuppressant TDM using online solid-phase extraction (SPE) and accurate-mass full scan-single ion monitoring (FS-SIM) data acquisition mode.
UNASSIGNED: LC-MS analysis was performed on a TLX-2 multi-channel HPLC with a Q-Exactive Plus mass spectrometer. TurboFlow online SPE was used for sample clean up. The accurate-mass MS was set to positive electrospray ionization mode with FS-SIM for quantitation of tacrolimus, sirolimus, everolimus, and cyclosporine A. MS2 fragmentation pattern was used for compound confirmation.
UNASSIGNED: The method was validated in terms of precision, analytical bias, limit of quantitation, linearity, carryover, sample stability, and interference. Quantitation of tacrolimus, sirolimus, everolimus, and cyclosporine A correlated well with results from an independent reference laboratory (r = 0.926-0.984).
UNASSIGNED: Accurate-mass FS-SIM can be successfully utilized for immunosuppressant TDM with good correlation with results generated by standard methods. TurboFlow online SPE allows for a simple \"protein crash and shoot\" sample preparation protocol. Compared to traditional MRM, analyte quantitation by FS-SIM facilitates a streamlined assay optimization process.