Liver transplantation is the definitive treatment for end-stage liver failure, but the scarcity of donor organs remains a significant challenge. Leveraging organs from extended criteria donors (ECD) offers a potential avenue to address worldwide shortages, though these organs are more susceptible to post-reperfusion injury. This study explores the use of normothermic ex vivo liver perfusion (NEVLP) as a method for organ preservation – an approach that sustains liver metabolism and facilitates pre-transplant assessments of organ viability via bile analysis. The focal point of this study revolves on the development of analytical methods for determining the bile acid profile throughout the peritransplantation period as a potential indicator of liver function and viability. The study optimized and validated a high-throughput analytical method to quantify selected bile acids in bile samples using a thin-film microextraction-liquid chromatography-mass spectrometry (TFME-LC-MS) platform. Furthermore, it introduced a solid-phase microextraction-microfluidic open interface-mass spectrometry (SPME-MOI-MS) method for rapid direct analysis of bile acid isobar groups. In the animal study, discernible variations in the concentrations of specific bile acids were observed between donors after circulatory death (DCD) and heart-beating donors (HBD), particularly following normothermic perfusion and reperfusion. Noteworthy fluctuations in individual bile acid concentrations were observed throughout the entire organ transplantation process, with taurocholic acid (TCA), glycocholic acid (GCA), and glycochenodeoxycholic acid (GCDCA) emerging as promising indicators of organ quality. The efficacy of the SPME-MOI-MS platform in corroborating these trends highlights its potential for real-time bile acid analysis during liver transplantation procedures. Our findings underscore the efficacy of NEVLP in tandem with advanced bile acid analysis methods as a reliable strategy for pre-transplant assessments of organ viability, potentially increasing the use of ECD organs and reducing organ shortages. The ability to monitor bile acid profiles in real-time provides crucial insights into liver function and ischemic injury, making significant strides in improving transplant outcomes and patient survival rates.

中文翻译：

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最近基于固相微萃取的分析方法，用于在常温离体肝脏灌注的肝移植物的移植前评估中分析胆汁胆汁酸


肝移植是终末期肝衰竭的最终治疗方法，但供体器官的稀缺仍然是一个重大挑战。利用扩展标准供体 (ECD) 的器官提供了解决全球器官短缺的潜在途径，尽管这些器官更容易受到再灌注后损伤。这项研究探索了使用常温离体肝脏灌注（NEVLP）作为器官保存的方法——一种维持肝脏代谢并通过胆汁分析促进器官活力移植前评估的方法。这项研究的重点是开发分析方法，用于确定整个围移植期的胆汁酸谱，作为肝功能和活力的潜在指标。该研究优化并验证了一种高通量分析方法，使用薄膜微萃取-液相色谱-质谱 (TFME-LC-MS) 平台对胆汁样品中选定的胆汁酸进行定量。此外，它还引入了固相微萃取-微流控开放界面-质谱（SPME-MOI-MS）方法，用于快速直接分析胆汁酸同量异位素基团。在动物研究中，观察到循环死亡（DCD）供体和心跳供体（HBD）之间特定胆汁酸浓度的明显变化，特别是在常温灌注和再灌注后。在整个器官移植过程中，观察到个体胆汁酸浓度的显着波动，其中牛磺胆酸（TCA）、甘氨胆酸（GCA）和甘氨鹅脱氧胆酸（GCDCA）成为有前景的器官质量指标。 SPME-MOI-MS 平台在证实这些趋势方面的功效凸显了其在肝移植过程中进行实时胆汁酸分析的潜力。我们的研究结果强调了 NEVLP 与先进胆汁酸分析方法相结合的功效，作为移植前评估器官活力的可靠策略，有可能增加 ECD 器官的使用并减少器官短缺。实时监测胆汁酸分布的能力为肝功能和缺血性损伤提供了重要的见解，在改善移植结果和患者存活率方面取得了重大进展。