Development and application of a fast ultra-high performance liquid chromatography-trapped ion mobility mass spectrometry method for untargeted lipidomics
Development and application of a fast ultra-high performance liquid chromatography-trapped ion mobility mass spectrometry method for untargeted lipidomics
复制标题
DOI：
10.1016/j.chroma.2022.463124
复制
发表时间：
2022-05-12
影响因子：
4.1
通讯作者：
Campiglia, Pietro
中科院分区：
文献类型：
--
作者：
Merciai, Fabrizio;Musella, Simona;Campiglia, Pietro
生物样本（如血浆）中脂质谱的改变反映了其体内平衡的异常，并为疾病理解提供了关键信息。需要快速分析方法来突出血脂谱的变化并提供快速结果。本研究建立了一种快速反相超高效液相色谱-离子迁移质谱（RP-UHPLC-TIMS-MS）非靶向脂质组学分析方法。一种短的、窄口径的全多孔颗粒CSH柱（50 mm × 2.1 mm，1.7 μ m），通过选择合适的流速、温度和梯度条件，总分析时间从20分钟减少到4分钟。TIMS以平行累积串联裂解模式（PASEF）操作，其允许选择MS/MS的多种前体并分离共-基于它们不同的迁移率洗脱脂质。通过基于规则的方法进行脂质注释，与LipidBlast光谱库和手动数据管理进行比较，考虑类别特异性碎片模式，准确质量，加合物形式，RP中的保留行为以及比较其碰撞截面（CCS）值以增加置信度。来自21个亚类的306种独特脂质从20 μ L血浆中注释，而它们的浓度通过类别特异性氘代内标物估计。分析方法进行了验证，并最终应用于阐明一个小队列的肌萎缩侧索硬化症（ALS）患者的血脂谱的改变。单变量和多变量统计证明了与对照患者的显著差异，特别是醚连接脂质（PC-O、PE-O、PE-P和LPC-O）、鞘脂（神经酰胺）和三酰甘油水平，表明这种快速方法在提供更长（>= 15 min）非靶向UHPLC-HRMS方法的准确和快速结果方面的有用性。(c)2022爱思唯尔有限公司版权所有。
The alteration of lipid profile in biological specimens, such as plasma, mirrors abnormalities in their homeostasis and offers pivotal information for disease comprehension. Fast analytical methods are needed to highlight changes in plasma lipid profile and deliver rapid results. In this study we developed a fast reversed phase ultra high performance liquid chromatography-trapped ion mobility mass spectrometry (RP-UHPLC-TIMS-MS) method for untargeted lipidomics. A short, narrow-bore fully porous particle CSH column (50 mm x 2.1 mm, 1.7 mu m) was used, and by selecting appropriate flow rate, temperature and gradient conditions, the total analysis time was reduced from 20 to 4 min. TIMS was operated in parallel accumulation serial fragmentation mode (PASEF) which allowed to select multiple precursors for MS/MS and separate co-eluting lipids based on their different mobility. Lipid annotation was performed by rule-based approach, comparison with LipidBlast spectral library and manual data curation, by taking into account class-specific fragmentation pattern, accurate mass, adduct form, retention behavior in RP and comparison of their collision cross-section (CCS) values for increased confidence. 306 unique lipids from 21 subclasses were annotated from 20 mu L of plasma, while their concentration was estimated by class-specific deuterated internal standards. The analytical method was validated and finally applied to elucidate the alteration of plasma lipid profiles in a small cohort of amyotrophic lateral sclerosis (ALS) patients. Univariate and multivariate statistics evidenced significant differences with respect to control patients, particularly in the levels of ether linked lipids (PC-O, PE-O, PE-P and LPC-O), sphingolipids (Ceramides), and triacylglycerols, showing the usefulness of this fast approach in providing accurate and rapid results with respect to longer ( >= 15 min) untargeted UHPLC-HRMS methods. (c) 2022 Elsevier B.V. All rights reserved.