利用SWATH質譜技術尋找小鼠肝臟中受聚苯乙烯塑膠微粒誘導之差異性代謝體

Abstract

近年來，塑膠微粒污染引起了國際的關注。 有報導稱塑膠微粒可能對人類產生肝毒性。代謝體學是評估此類污染物威脅的有力策略，可以更直接地反映其表型。 在本研究中，我們應用液相層析-串聯質譜 (LC-MS/MS) 和SWATH 數據獲取模式評估暴露於聚苯乙烯塑膠微粒的小鼠肝臟差異性代謝體學。在差異性代謝體學分析之前， 8 種不同 LC 條件的覆蓋範圍透過使用混合樣品，並使用代謝物標準混合物評估兩個親水相互作用液相層析 (HILIC) 柱的定量性能。結果表明，Amide管柱在ESI(+)和ESI(-)模式下最適合小分子分析，而流動相中含有0.1% FA的PFP柱最適合ESI(-)模式的磷脂分析，總共能夠鑑定923種代謝物。 LC-SWATH-MS 發現了 71 種差異代謝物，並且發現嘌呤和嘧啶代謝、氨基酸代謝和磷脂代謝受到聚苯乙烯塑膠微粒的干擾。這項研究提供了一個通用的代謝體學圖譜，可以揭示聚苯乙烯塑膠微粒暴露對小鼠肝臟的影響，這是血清化學分析和組織學分析無法給出的。

Microplastic (MP) pollution has gained international attention in recent years. It has also been reported that MPs could induce hepatotoxic in humans. Metabolomics is a powerful strategy for evaluating the threat of such pollutants, which can reflect the phenotype in a more direct way. In this study, we applied liquid chromatography-tandem mass spectrometry (LC-MS/MS) with sequential window acquisition of all theoretical mass spectra (SWATH) data acquisition mode to assess the differential metabolomics in mouse liver exposed to polystyrene MPs. Before differential metabolomics analysis, the coverage of 8 different chromatographic conditions was evaluated using the pooled sample, and the quantitative performance of the two hydrophilic interaction liquid chromatography (HILIC) columns was evaluated using metabolite standard mixtures. The results showed that the amide column was the best for small molecule analysis in the ESI(+) and the ESI(-) mode and the PFP column with 0.1% FA in the mobile phase was the best for phospholipids analysis in the ESI(-) mode. By combining the selected conditions, our method can identify 923 metabolites in summary. There were 70 differential metabolites discovered by the LC-SWATH-MS and the purine and pyrimidine metabolism, amino acid metabolism, and phospholipid metabolism were found to be disturbed by the PS-MPs. This study provides a universal metabolomics profile that can reveal the effects of PS-MPs exposure on the mouse liver that cannot be given by serum chemical analysis and histological analysis.