Strategy for Comprehensive Identification of Acylcarnitines Based on Liquid Chromatography-High-Resolution Mass Spectrometry
Strategy for Comprehensive Identification of Acylcarnitines Based on Liquid Chromatography-High-Resolution Mass Spectrometry
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基于液相色谱-高分辨率质谱的酰基肉碱综合鉴定策略
DOI：
10.1021/acs.analchem.7b05471
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发表时间：
2018
影响因子：
7.4
通讯作者：
Guowang Xu
中科院分区：
文献类型：
--
作者：
Di Yu;Lina Zhou;Qiuhui Xuan;Lichao Wang;Xinjie Zhao;Xin Lu;Guowang Xu
肉毒碱在脂肪酸氧化和支链氨基酸代谢中起重要作用。酰基肉毒碱代谢紊乱与许多疾病的发生、发展有关。对酰基肉毒碱进行全面的鉴定，有利于其靶向检测、后续疾病鉴别诊断和可能机制的研究。在这项研究中，我们开发了一种新的策略，以确定尽可能多的酰基肉毒碱，尽可能基于液相色谱-高分辨率质谱（LC-HRMS）。层-层渐进策略首先整合初始全扫描MS/数据依赖MS/MS监测（ddMS 2）采集和随后的并行反应监测（PRM）以分析合并的生物样品。还确定了733种可能的酰基肉毒碱，含有特征性的高分辨率MS/MS特征。此外，准确的质量，保留规则，和HRMS/MS信息被用来定义亚类和预测未检测到的酰基肉毒碱同系物在每个亚类，导致更多的酰基肉毒碱，我们新构建的数据库。结果，数据库中包含758个酰基肉毒碱，具有精确的质量，保留时间和MS/MS信息，这是迄今为止报道的酰基肉毒碱的最全面的列表。应用这个数据库，241，515，和222酰基肉毒碱在人血浆，人尿和大鼠肝组织中被快速和可靠地注释。这种新的策略能够大规模鉴定酰基肉毒碱，类似的方法也可用于鉴定其他代谢产物。
Carnitines play important roles in fatty acid oxidation and branched chain amino acid metabolism. The disturbance of acylcarnitines is associated with occurrence and development of many diseases. Comprehensive acylcarnitine identification can greatly benefit their targeted detection, following disease differential diagnosis and possible mechanism study. In this study, we developed a novel strategy to identify as many acylcarnitines as possible based on liquid chromatography–high-resolution mass spectrometry (LC–HRMS). The layer–layer progressive strategy first integrated the initial full scan MS/data-dependent MS/MS monitoring (ddMS2) acquisition and the following parallel reaction monitoring (PRM) to analyze a pooled biological sample. Also 733 possible acylcarnitines were identified containing characteristic high-resolution MS/MS features. Further, accurate mass, retention rules, and HRMS/MS information were used to define subclasses and predict undetected acylcarnitine homologues in each subclass, leading to more acylcarnitines to our newly constructed database. As a result, 758 acylcarnitines were contained in the database, having exact mass, retention time, and MS/MS information, which is the most comprehensive list of acylcarnitines reported to date. Applying this database, 241, 515, and 222 acylcarnitines were rapidly and reliably annotated in human plasma, human urine, and rat liver tissue. This novel strategy enables large-scale identification of acylcarnitines, and a similar method can also be used for identification of other metabolites.