Nazdrajić Emir, Tascon Marcos, Rickert Daniel A, Gómez-Ríos German A, Kulasingam Vathany, Pawliszyn Janusz B
Department of Chemistry, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
Department of Laboratory Medicine and Pathology, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Clinical Biochemistry, University Health Network, Toronto, ON M5G 2C4, Canada.
Anal Chim Acta. 2021 Feb 1;1144:53-60. doi: 10.1016/j.aca.2020.11.056. Epub 2020 Dec 4.
Immunosuppressive drugs are administered to decrease immune system activity (e.g. of patients undergoing solid organ transplant). Concentrations of immunosuppressive drugs (ISDs) in circulating blood must be closely monitored during the period of immunosuppression therapy due to adverse effects that take place when concentration levels fall outside of the very narrow therapeutic concentration range of these drugs. This study presents the rapid determination of four relevant immunosuppressive drugs (tacrolimus, sirolimus, everolimus, and cyclosporine A) in whole human blood by directly coupling solid-phase microextraction to mass spectrometry via the microfluidic open interface (Bio-SPME-MOI-MS/MS). The BioSPME-MOI-MS/MS method offers ≤ 10% imprecision of in-house prepared quality controls over a 10-day period, ≤ 10% imprecision of ClinCal® Recipe calibrators over a three-day period, and single total turnaround time of ∼ 60 min (4.5 min for high throughput). The limits of quantification were determined to be 0.8 ng mL for tacrolimus, 0.7 ng mL sirolimus, 1.0 ng mL for everolimus, and 0.8 ng mL for cyclosporine. The limits of detection were determined to be 0.3 ng mL for tacrolimus, 0.2 ng mL for sirolimus, 0.3 ng mL for everolimus, and 0.3 ng mL for cyclosporine A. The R values for all analytes were above 0.9992 with linear dynamic range from 1.0 mL to 50.0 ng mL for tacrolimus, sirolimus, and everolimus while from 2.5 ng mL to 500.0 ng mL for cyclosporine A. To further evaluate the performance of the present method, 95 residual whole blood samples of tacrolimus and sirolimus from patients undergoing immunosuppression therapy were used to compare the Bio-SPME-MOI-MS/MS method against a clinically validated reference method based on chemiluminescent microparticle immunoassay, showing acceptable results. Our results demonstrated that Bio-SPME-MOI-MS/MS can be considered as a suitable alternative to existing methods for the determination of immunosuppressive drugs in whole blood providing faster analysis, better selectivity and sensitivity, and a wider dynamic range than current existing approaches.
免疫抑制药物用于降低免疫系统活性（如实体器官移植患者）。在免疫抑制治疗期间，必须密切监测循环血液中免疫抑制药物（ISDs）的浓度，因为当浓度水平超出这些药物非常狭窄的治疗浓度范围时会产生不良反应。本研究介绍了通过微流控开放接口（Bio-SPME-MOI-MS/MS）将固相微萃取直接与质谱联用，快速测定全血中四种相关免疫抑制药物（他克莫司、西罗莫司、依维莫司和环孢素A）的方法。BioSPME-MOI-MS/MS方法在10天内对内部制备的质量控制品的不精密度≤10%，在三天内对ClinCal®配方校准品的不精密度≤10%，单次总周转时间约为60分钟（高通量时为4.5分钟）。他克莫司的定量限为0.8 ng/mL，西罗莫司为0.7 ng/mL，依维莫司为1.0 ng/mL，环孢素为0.8 ng/mL。他克莫司的检测限为0.3 ng/mL，西罗莫司为0.2 ng/mL，依维莫司为0.3 ng/mL，环孢素A为0.3 ng/mL。所有分析物的R值均高于0.9992，他克莫司、西罗莫司和依维莫司的线性动态范围为1.0 ng/mL至50.0 ng/mL，环孢素A为2.5 ng/mL至500.0 ng/mL。为了进一步评估本方法的性能，使用了95份免疫抑制治疗患者的他克莫司和西罗莫司残留全血样本，将Bio-SPME-MOI-MS/MS方法与基于化学发光微粒子免疫测定的临床验证参考方法进行比较，结果令人满意。我们的结果表明，Bio-SPME-MOI-MS/MS可被视为全血中免疫抑制药物测定现有方法的合适替代方法，与现有方法相比，它提供了更快的分析速度、更好的选择性和灵敏度以及更宽的动态范围。
