Enhanced Lipidomics Analysis of Breast Cancer Cells Using Three-phase Liquid Extraction and Ultra High-performance Liquid Chromatography Coupled With Quadrupole Time-of-Flight Tandem Mass Spectrometry

IF 2.8 3区 工程技术 Q2 CHEMISTRY, ANALYTICAL Journal of separation science Pub Date : 2024-11-04 DOI:10.1002/jssc.70014
Binhong He, Fengying Ye, Jieqing Feng, Ting Zhou
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Abstract

Lipid extraction of complex biological samples is essential for high-quality data in liquid chromatography-mass spectrometry (LC-MS)-based lipidomics. This study introduces a three-phase liquid extraction (3PLE)-ultra-high-performance LC coupled with quadrupole time-of-flight tandem MS method. This method was successfully applied to lipidomics analysis of breast cancer cells, including highly metastatic MDA-MB-231 and slightly metastatic MCF7 cells. The 3PLE method employed an n-hexane/methyl tert-butyl ether/acetonitrile/water solvent system that formed one aqueous and two organic phases. Neutral and polar lipids were enriched in the upper and middle organic phases, respectively, and combined for detection, thereby reducing analysis time. Compared with the Bligh and Dyer method, 3PLE achieved higher sensitivity and detected more features, with over a 50% increase in the relative abundance of nearly 50% of the differential lipids. In total, 21 differential lipids were identified in the MDA-MB-231 group and 22 in the MCF7 group compared to normal breast epithelial cells (MCF10A). Pathway analysis suggested that lipid changes in breast cancer cells were associated with glycerophospholipid metabolism, arachidonic acid metabolism, sphingolipid metabolism, and linoleic acid metabolism. The study presents a highly efficient lipidomics method, providing a scientific foundation for understanding breast cancer pathogenesis and aiding in diagnosis.

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利用三相液相萃取和超高效液相色谱-四极杆飞行时间串联质谱法加强乳腺癌细胞脂质组学分析
在基于液相色谱-质谱联用技术(LC-MS)的脂质组学研究中,复杂生物样品的脂质提取对于获得高质量的数据至关重要。本研究介绍了一种三相液体萃取(3PLE)-超高效液相色谱与四极杆飞行时间串联质谱联用方法。该方法被成功应用于乳腺癌细胞的脂质组学分析,包括高转移性MDA-MB-231和轻微转移性MCF7细胞。3PLE 方法采用正己烷/甲基叔丁基醚/乙腈/水溶剂系统,形成一个水相和两个有机相。中性脂质和极性脂质分别富集在上层和中层有机相中,并合并在一起进行检测,从而缩短了分析时间。与 Bligh 和 Dyer 方法相比,3PLE 的灵敏度更高,检测到的特征更多,近 50%的差异脂质的相对丰度提高了 50%以上。与正常乳腺上皮细胞(MCF10A)相比,MDA-MB-231 组共鉴定出 21 种差异脂质,MCF7 组鉴定出 22 种差异脂质。通路分析表明,乳腺癌细胞的脂质变化与甘油磷脂代谢、花生四烯酸代谢、鞘脂代谢和亚油酸代谢有关。该研究提出了一种高效的脂质组学方法,为了解乳腺癌的发病机制和辅助诊断提供了科学依据。
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来源期刊
Journal of separation science
Journal of separation science 化学-分析化学
CiteScore
6.30
自引率
16.10%
发文量
408
审稿时长
1.8 months
期刊介绍: The Journal of Separation Science (JSS) is the most comprehensive source in separation science, since it covers all areas of chromatographic and electrophoretic separation methods in theory and practice, both in the analytical and in the preparative mode, solid phase extraction, sample preparation, and related techniques. Manuscripts on methodological or instrumental developments, including detection aspects, in particular mass spectrometry, as well as on innovative applications will also be published. Manuscripts on hyphenation, automation, and miniaturization are particularly welcome. Pre- and post-separation facets of a total analysis may be covered as well as the underlying logic of the development or application of a method.
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