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Simultaneous determination of tryptophan and its 31 catabolites in mouse tissues by polarity switching UHPLC-SRM-MS
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2018-12-01 , DOI: 10.1016/j.aca.2018.02.026 Guan-Yuan Chen 1 , Wei Zhong 1 , Zhanxiang Zhou 2 , Qibin Zhang 3
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2018-12-01 , DOI: 10.1016/j.aca.2018.02.026 Guan-Yuan Chen 1 , Wei Zhong 1 , Zhanxiang Zhou 2 , Qibin Zhang 3
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Tryptophan (TRP) and its catabolites have attracted a lot of attention because of their clinical significance to human health. Recently, microbiome-gut-brain axis was found to have links to many diseases based on the imbalance of TRP catabolism. By using ultra-high performance liquid chromatography coupled to electrospray ionization triple quadrupole mass spectrometry, we present a rapid, robust and comprehensive method to determine 31 TRP catabolites covering three major pathways - kynurenic, serotonergic and bacterial degradation - within 5 min. Polarity switching was employed to analyze catabolites in both ionization modes simultaneously for greatly improved analytical throughput. The intra-day and inter-day precision were 0.5-15.8% and 1.5-16.7%, respectively. Accuracy was between 75.8 and 126.9%. The developed method was applied to study the tissue level of TRP catabolites in the liver, ileum, ileal contents, brain and plasma samples from 8 mice, and clear differences in the distribution of TRP catabolites were observed in different tissues. Ratios of key catabolites to TRP were used to evaluate the activities of specific enzyme and pathway in respective tissues. This method has potential in high throughput analysis of TRP catabolites in biological matrices, which can facilitate understanding the influence of TRP catabolites on microbiome-gut-brain axis and on human health.
中文翻译：
极性转换 UHPLC-SRM-MS 同时测定小鼠组织中色氨酸及其 31 种分解代谢物
色氨酸（TRP）及其分解代谢物因其对人类健康的临床意义而受到广泛关注。最近，基于 TRP 分解代谢的不平衡，发现微生物组-肠-脑轴与许多疾病存在联系。通过使用超高效液相色谱与电喷雾电离三重四极杆质谱联用，我们提出了一种快速、稳健且全面的方法，可在 5 分钟内测定涵盖三种主要途径（犬尿酸、血清素能和细菌降解）的 31 种 TRP 分解代谢物。采用极性切换同时分析两种电离模式下的分解代谢物，从而大大提高了分析通量。日内和日间精度分别为0.5-15.8%和1.5-16.7%。准确度在 75.8% 至 126.9% 之间。应用所开发的方法研究了8只小鼠的肝脏、回肠、回肠内容物、脑和血浆样本中TRP分解代谢物的组织水平，并观察到不同组织中TRP分解代谢物的分布存在明显差异。关键分解代谢物与 TRP 的比率用于评估各个组织中特定酶和途径的活性。该方法在生物基质中 TRP 分解代谢物的高通量分析方面具有潜力，有助于了解 TRP 分解代谢物对微生物组-肠-脑轴和人类健康的影响。
更新日期：2018-12-01
中文翻译：
极性转换 UHPLC-SRM-MS 同时测定小鼠组织中色氨酸及其 31 种分解代谢物
色氨酸（TRP）及其分解代谢物因其对人类健康的临床意义而受到广泛关注。最近，基于 TRP 分解代谢的不平衡，发现微生物组-肠-脑轴与许多疾病存在联系。通过使用超高效液相色谱与电喷雾电离三重四极杆质谱联用，我们提出了一种快速、稳健且全面的方法，可在 5 分钟内测定涵盖三种主要途径（犬尿酸、血清素能和细菌降解）的 31 种 TRP 分解代谢物。采用极性切换同时分析两种电离模式下的分解代谢物，从而大大提高了分析通量。日内和日间精度分别为0.5-15.8%和1.5-16.7%。准确度在 75.8% 至 126.9% 之间。应用所开发的方法研究了8只小鼠的肝脏、回肠、回肠内容物、脑和血浆样本中TRP分解代谢物的组织水平，并观察到不同组织中TRP分解代谢物的分布存在明显差异。关键分解代谢物与 TRP 的比率用于评估各个组织中特定酶和途径的活性。该方法在生物基质中 TRP 分解代谢物的高通量分析方面具有潜力，有助于了解 TRP 分解代谢物对微生物组-肠-脑轴和人类健康的影响。
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