UHPLC-TIMS-PASEF^®-MS for Lipidomics: From Theory to Practice.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4334-1_12

Vanna Denti, Simone Serrao, Eleonora Bossi, Giuseppe Paglia

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引用次数: 0

Abstract

Trapped ion mobility spectrometry (TIMS) using parallel accumulation serial fragmentation (PASEF^®) is an advanced analytical technique that offers several advantages in mass spectrometry (MS)-based lipidomics. TIMS provides an additional dimension of separation to mass spectrometry and accurate collision cross-section (CCS) measurements for ions, aiding in the structural characterization of molecules. This is especially valuable in lipidomics for identifying and distinguishing isomeric or structurally similar compounds. On the other hand, PASEF technology allows for fast and efficient data acquisition by accumulating ions in parallel and then serially fragmenting them. This accelerates the analysis process and improves throughput, making it suitable for high-throughput applications. Moreover, the combination of TIMS and PASEF reduces co-elution and ion coalescence issues, leading to cleaner and more interpretable mass spectra. This results in higher data quality and more confident identifications. In this chapter, a data-dependent TIMS-PASEF^® workflow for lipidomics analysis is presented.

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UHPLC-TIMS-PASEF®-MS用于脂质组学：从理论到实践

使用平行积累序列碎片（PASEF®）的捕获离子迁移率光谱（TIMS）是一种先进的分析技术，在质谱（MS）为基础的脂质组学中提供了几个优势。TIMS为质谱和离子的精确碰撞截面（CCS）测量提供了额外的分离维度，有助于分子的结构表征。这在鉴别和区分同分异构体或结构相似的化合物的脂质组学中特别有价值。另一方面，PASEF技术允许通过并行积累离子然后串行分割它们来快速有效地获取数据。这加快了分析过程，提高了吞吐量，使其适用于高吞吐量应用。此外，TIMS和PASEF的结合减少了共洗脱和离子聚结问题，导致更清洁和更可解释的质谱。这将导致更高的数据质量和更自信的识别。在本章中，提出了用于脂质组学分析的数据依赖的TIMS-PASEF®工作流程。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.