School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Guangzhou, China.
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
Biomed Chromatogr. 2021 Feb;35(2):e4985. doi: 10.1002/bmc.4985. Epub 2020 Nov 17.
Gut microbial phenylalanine, tyrosine, and tryptophan metabolites are closely linked to various diseases. Monitoring the alterations of the related metabolites is vital to facilitate the understanding of pathophysiology of diseases. Herein, a rapid and sensitive assay based on LC-tandem mass spectrometry has been developed to analyze 20 gut microbial metabolites derived from phenylalanine, tyrosine, and tryptophan in rat serum, urine, and faeces. These microbial-derived metabolites were separated on a phenyl-hexyl column and simultaneously determined in a single run of 8 min. The detection limit for analytes ranged between 1.08 and 32.4 ng/mL. All calibration curves exhibited good linear relationships (R ≥ 0.9982). Intra- and inter-assay precision values were below 15% and accuracies ranged from 85% to 115% for all analytes. The selectivity, matrix effect, and recovery of this method were all satisfactory. The validated method was successfully applied to characterize the alterations of these metabolites in type 2 diabetes mellitus rat. In general, the developed assay is suitable for high-throughput monitoring of gut microbial phenylalanine, tyrosine, and tryptophan metabolites and provides a useful approach for exploring the mechanisms of microbial-derived metabolites in diseases.
肠道微生物衍生的苯丙氨酸、酪氨酸和色氨酸代谢物与各种疾病密切相关。监测相关代谢物的变化对于促进疾病病理生理学的理解至关重要。本文建立了一种基于 LC-串联质谱的快速灵敏检测方法，用于分析大鼠血清、尿液和粪便中 20 种源自苯丙氨酸、酪氨酸和色氨酸的肠道微生物代谢物。这些微生物衍生的代谢物在苯基-己基柱上分离，并在 8 分钟的单次运行中同时测定。分析物的检测限在 1.08 至 32.4ng/mL 之间。所有校准曲线均表现出良好的线性关系（R ≥ 0.9982）。所有分析物的内、日间精密度值均低于 15%，准确度在 85%至 115%之间。该方法的选择性、基质效应和回收率均令人满意。所建立的方法成功应用于表征 2 型糖尿病大鼠这些代谢物的变化。总之，该方法适用于高通量监测肠道微生物衍生的苯丙氨酸、酪氨酸和色氨酸代谢物，为探索疾病中微生物衍生代谢物的机制提供了一种有用的方法。
