Liver enzymes and transporters play an essential role in xenobiotic metabolism, distribution and elimination. Pre-clinical safety assessment relies on studies on animal models, including the (mini)pig. The pig shares many anatomical and physiological characteristics with humans, and there is currently a gap in information about porcine metabolism and disposition pathways and their similarities and differences from human ones.

Three different sample preparation methods (filter-aided sample preparation (FASP), enhanced FASP (eFASP) and in-solution sample preparation) were used to prepare porcine liver tissue (two samples) for proteomic analysis. The analysis relied on rapid-separation liquid chromatography coupled to Orbitrap mass spectrometry in data-dependent acquisition mode. MASCOT was used for identification and relative label-free quantification was based on spectral counting.

The three sample preparation methods provided complementary results, allowing characterisation of approximately 70 pharmacologically relevant proteins. The main quantified proteins included 16 cytochrome P450 (CYP) enzymes, 5 UGT enzymes, and 11 transporters. In addition, 20 Phase I and 14 Phase II enzymes were also characterised. Inter-operator differences were negligible and the pig liver pies for CYP, UGT and efflux transporter proteins were established. Human homologues of the quantified CYP, UGT and transporter proteins were identified.

肝酶和转运蛋白在异源生物的代谢，分布和消除中起着至关重要的作用。临床前安全性评估依赖于对动物模型的研究，包括（小型）猪。猪与人具有许多解剖和生理特征，目前关于猪的新陈代谢和处置途径以及它们与人的异同的信息尚存在差距。

三种不同的样品制备方法（过滤辅助样品制备（FASP），增强型FASP（eFASP）和溶液中样品制备）用于制备猪肝组织（两个样品）进行蛋白质组学分析。该分析依赖于快速分离液相色谱与Orbitrap质谱联用的数据相关采集模式。MASCOT用于鉴定，相对无标记的定量基于光谱计数。

三种样品制备方法提供了互补的结果，可表征约70种药理相关蛋白。定量的主要蛋白质包括16种细胞色素P450（CYP）酶，5种UGT酶和11种转运蛋白。此外，还鉴定了20种I相和14种II期酶。操作者之间的差异可忽略不计，并建立了猪肝中CYP，UGT和外排转运蛋白的表达。鉴定了定量CYP，UGT和转运蛋白的人同源物。