Zhou Juntuo, Li Yuhua, Chen Xi, Zhong Lijun, Yin Yuxin
Department of Pathology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China.
Medical and Health Analytical Center, Peking University Health Science Center, Beijing 100191, China.
Talanta. 2017 Mar 1;164:128-136. doi: 10.1016/j.talanta.2016.11.048. Epub 2016 Nov 21.
Untargeted metabolomic profiling has been widely used in recent years. However, the low reproducibility of the data-dependent acquisition (DDA) strategy presents a major bottleneck that considerably limits the reliability of metabolomic studies in biological and clinical research. The data-independent acquisition (DIA) strategy is proposed to solve the above-mentioned problem, and it is gaining popularity. This paper presents a novel approach for performing metabolomic analysis using an untargeted, liquid chromatography-data independent-mass spectrometry (LC-DIA-MS) strategy on a quadrupole-Orbitrap platform. Using chemical standards and metabolites extracted from serum samples, we optimized the LC-DIA-MS parameters to analyze hydrophilic metabolites and lipids. The quantitative performance and analytical reliability were evaluated, and the performances of DIA, DDA, and all-ion fragmentation mode were compared. Finally, as a proof of concept, we applied the constructed DIA workflow to a comparative metabolomic study of papillary thyroid carcinoma (TC) serum samples. Several metabolites, including carnitine, trimethylamine N-oxide, and some amino acids, significantly changed between patients and healthy controls. This study demonstrated the feasibility and advantage of the DIA strategy on untargeted metabolomic analysis for biological study and clinical biomarker screening.
近年来，非靶向代谢组学分析已被广泛应用。然而，数据依赖型采集（DDA）策略的低重现性成为一个主要瓶颈，极大地限制了代谢组学研究在生物和临床研究中的可靠性。为了解决上述问题，人们提出了数据非依赖型采集（DIA）策略，且该策略正日益受到欢迎。本文介绍了一种在四极杆-轨道阱平台上使用非靶向液相色谱-数据非依赖型质谱（LC-DIA-MS）策略进行代谢组学分析的新方法。利用化学标准品和从血清样本中提取的代谢物，我们优化了LC-DIA-MS参数以分析亲水性代谢物和脂质。评估了定量性能和分析可靠性，并比较了DIA、DDA和全离子碎裂模式的性能。最后，作为概念验证，我们将构建的DIA工作流程应用于甲状腺乳头状癌（TC）血清样本的比较代谢组学研究。包括肉碱、氧化三甲胺和一些氨基酸在内的几种代谢物在患者和健康对照之间有显著变化。本研究证明了DIA策略在非靶向代谢组学分析用于生物学研究和临床生物标志物筛选方面的可行性和优势。
