Tao Xin, Liu Jia-Yue, Zhou Jun-Yi, Dai Jiang-Kun, Xiao Zeyu, Li Hou-Kai, Wan Jian-Bo
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macao SAR 999078, China.
Collaborative Translational Medicine Collaborative Innovation Center, Department of Pharmacology and Chemical Biology, Institute of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Anal Chem. 2024 Dec 31;96(52):20414-20424. doi: 10.1021/acs.analchem.4c04037. Epub 2024 Dec 19.
Numerous bioactive compounds containing carbonyl groups, including aldehydes and ketones, are widely acknowledged as potential biomarkers for several diseases and are implicated in the development of metabolic disorders. However, the detection of carbonyl metabolites is hindered by challenges, such as poor ionization efficiency, low biological concentration, instability, and complexity of the sample matrix. To overcome these limitations, we developed a Girard derivatization-based enrichment (GDBE) strategy for capturing and comprehensively profiling carbonyl metabolites in biological samples. A functionalized resin, named carbonyl capture and reporter-ion installation (CCRI) resins, was synthesized to selectively capture carbonyl metabolites via a Girard reaction. After unwanted metabolites were removed, the hydrazone derivatives were cleaved from the solid-phase resins and subjected to LC-MS analysis. The proposed GDBE strategy exhibits exceptional selectivity for capturing and enriching carbonyl metabolites. Moreover, this method surpasses current detection limits by enhancing the MS sensitivity and facilitating structural characterization of hydrazone derivatives by a specific MS/MS fragmentation signature. Using the GDBE method, 957 potential carbonyl metabolites were successfully identified in liver tissue from alcohol-fed mice. Among them, 76 carbonyl metabolites were annotated, indicating the potential of this strategy for the efficient nontargeted profiling of the carbonyl submetabolome in complex biological samples.
许多含有羰基的生物活性化合物，包括醛类和酮类，被广泛认为是多种疾病的潜在生物标志物，并与代谢紊乱的发展有关。然而，羰基代谢物的检测受到诸多挑战的阻碍，如电离效率低、生物浓度低、稳定性差以及样品基质复杂等。为克服这些限制，我们开发了一种基于吉拉德衍生化的富集（GDBE）策略，用于捕获和全面分析生物样品中的羰基代谢物。合成了一种功能化树脂，名为羰基捕获与报告离子安装（CCRI）树脂，通过吉拉德反应选择性地捕获羰基代谢物。去除不需要的代谢物后，腙衍生物从固相树脂上裂解下来并进行液相色谱-质谱分析。所提出的GDBE策略在捕获和富集羰基代谢物方面表现出卓越的选择性。此外，该方法通过提高质谱灵敏度并借助特定的二级质谱碎片特征促进腙衍生物的结构表征，超越了当前的检测限。使用GDBE方法，在酒精喂养小鼠的肝脏组织中成功鉴定出957种潜在的羰基代谢物。其中，76种羰基代谢物得到注释，表明该策略在复杂生物样品中对羰基亚代谢组进行高效非靶向分析的潜力。
