Department of Chemistry - BMC, Science for Life Laboratory, Uppsala University, 75124 Uppsala, Sweden.
Department of Nutritional Sciences, School of Life Course and Population Sciences, Faculty of Life Sciences and Medicine, King's College London, London SE1 9NH, UK.
Anal Chem. 2023 Aug 22;95(33):12565-12571. doi: 10.1021/acs.analchem.3c02988. Epub 2023 Aug 8.
The human body has evolved to remove xenobiotics through a multistep clearance process. Non-endogenous metabolites are converted through a series of phase I and different phase II enzymes into compounds with higher hydrophilicity. These compounds are important for diverse research fields such as toxicology, nutrition, biomarker discovery, doping control, and microbiome metabolism. One of the challenges in these research fields has been the investigation of the two major phase II modifications, sulfation and glucuronidation, and the corresponding unconjugated aglycon independently. We have now developed a new methodology utilizing an immobilized arylsulfatase and an immobilized β-glucuronidase to magnetic beads for treatment of human urine samples. The enzyme activities remained the same compared to the enzyme in solution. The separate mass spectrometric investigation of each metabolite class in a single sample was successfully applied to obtain the dietary glucuronidation and sulfation profile of 116 compounds. Our new chemical biology strategy provides a new tool for the investigation of metabolites in biological samples with the potential for broad-scale application in metabolomics, nutrition, and microbiome studies.
人体通过多步清除过程进化出消除异生物的能力。非内源性代谢物通过一系列的 I 相和不同的 II 相酶转化为亲水性更高的化合物。这些化合物在毒理学、营养、生物标志物发现、兴奋剂控制和微生物组代谢等多个研究领域都很重要。这些研究领域的挑战之一是分别研究两种主要的 II 相修饰，磺化和葡萄糖醛酸化，以及相应的未结合的糖苷配基。我们现在开发了一种新的方法，利用固定化芳基硫酸酯酶和固定化β-葡萄糖醛酸酶处理人尿液样本。与溶液中的酶相比，酶活性保持不变。成功地将每种代谢物类别的单独质谱分析应用于单个样品中，以获得 116 种化合物的膳食葡萄糖醛酸化和磺化谱。我们的新化学生物学策略为研究生物样本中的代谢物提供了一种新工具，具有在代谢组学、营养和微生物组研究中广泛应用的潜力。
