Wang Aimin, Fan Yuting, Fu Jun, Song Fengrui, Liu Zhiqiang, Liu Shu
State Key Laboratory of Electroanalytical Chemistry, National Center of Mass Spectrometry in Changchun, Jilin Province Key Laboratory of Chinese Medicine Chemistry and Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, China.
School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei, 230029, China.
Anal Bioanal Chem. 2023 Oct;415(25):6345-6353. doi: 10.1007/s00216-023-04910-5. Epub 2023 Aug 25.
Metabolomics is a biochemical analysis tool for identifying metabolic phenotypes and used to reveal the pathogenic mechanisms of disease and to inform drug-targeted therapies. Carboxyl-containing metabolites (CCMs) account for an important proportion of the metabolome, but because of the diversity of physical and chemical properties of CCMs in biological samples, traditional liquid chromatography-mass spectrometry (LC-MS) targeted metabolome analysis methods cannot achieve simultaneous quantification of multiple types of CCMs. Therefore, we proposed for the first time a targeted metabolomics strategy using isoniazid derivatization combined with LC-MS/MS to simultaneously quantify 39 CCMs of 5 different types (short-chain fatty acids, amino acids, bile acids, phenylalanine and tryptophan metabolic pathway acids) with large polarity differences associated with Alzheimer's disease (AD) and significantly improve the detection coverage and sensitivity. The yields of isoniazid derivative CCMs were high and could guarantee the accuracy of CCM quantification. The LODs of CCMs increased significantly (1.25-2000-fold) after derivatization. The method showed good selectivity, intra-day and inter-day accuracies and precisions, and repeatability. There was no significant effect on the determination of CCMs in terms of matrix effect and recovery. CCMs showed good stability. And CCMs showed good stability under short-term storage and freeze-thaw cycles. At the same time, the regulatory effects of Schisandrae chinensis Fructus and Ginseng Radix et Rhizoma (SG) herb pair on CCM metabolic disorders in feces, urine, serum, and the brain of AD rats were elucidated from the perspective of targeted metabolomics. In combination with pharmacodynamic evaluation and gut microbiota analysis, the mechanism of SG herb pair on AD rats was comprehensively understood. In summary, this innovative isoniazid derivatization combined with a targeted metabolomics method has great potential for trace biological lineage analysis.
代谢组学是一种用于识别代谢表型的生化分析工具，可用于揭示疾病的致病机制并为药物靶向治疗提供依据。含羧基代谢物（CCMs）在代谢组中占重要比例，但由于生物样品中CCMs理化性质的多样性，传统的液相色谱-质谱联用（LC-MS）靶向代谢组分析方法无法实现多种类型CCMs的同时定量。因此，我们首次提出了一种靶向代谢组学策略，即利用异烟肼衍生化结合LC-MS/MS同时定量5种不同类型（短链脂肪酸、氨基酸、胆汁酸、苯丙氨酸和色氨酸代谢途径酸）、与阿尔茨海默病（AD）相关且极性差异较大的39种CCMs，并显著提高检测覆盖率和灵敏度。异烟肼衍生化CCMs的产率较高，可保证CCMs定量的准确性。衍生化后CCMs的检测限显著提高（1.25-2000倍）。该方法具有良好的选择性、日内和日间准确性及精密度以及重复性。在基质效应和回收率方面对CCMs的测定无显著影响。CCMs表现出良好的稳定性，在短期储存和冻融循环下也具有良好的稳定性。同时，从靶向代谢组学的角度阐明了五味子果实与人参（SG）药对对AD大鼠粪便、尿液、血清和脑中CCM代谢紊乱的调节作用。结合药效学评价和肠道微生物群分析，全面了解了SG药对对AD大鼠的作用机制。综上所述，这种创新的异烟肼衍生化结合靶向代谢组学方法在痕量生物谱系分析方面具有巨大潜力。
