{"title":"通过优化流动相改性剂和色谱柱选择,使用基于超高效液相色谱-质谱的方法提高非生物小鼠模型的肠道脂质体覆盖率","authors":"Habiba Selmi , Alesia Walker , Laurent Debarbieux , Philippe Schmitt-Kopplin","doi":"10.1016/j.jchromb.2024.124188","DOIUrl":null,"url":null,"abstract":"<div><p>Lipidomics is focusing on the screening of lipid species in complex mixtures using mass spectrometry-based approaches. In this work, we aim to enhance the intestinal lipidome coverage within the Oligo-Mouse-Microbiota (OMM<sup>12</sup>) colonized mouse model by testing eight mobile phase conditions on five reversed-phase columns. Our selected mobile phase modifiers included two ammonium salts, two concentrations, and the addition of respective acids at 0.1 %. We compared two columns with hybrid surface technology, two with ethylene bridged hybrid technology and one with core–shell particles. Best performance was attained for standards and intestinal lipidome, using either ammonium formate or acetate in ESI(+) or ammonium acetate in ESI(−) for all column technologies. Notably, a concentration of 5 mM ammonium salt showed optimal results for both modes, while the addition of acids had a negligible effect on lipid ionization efficiency. The HST BEH C18 column improved peak width and tailing factor parameters compared to other technologies. We achieved the highest lipid count in colon and ileum content, including ceramides, phosphatidylethanolamines and phosphatidylcholines, when using 5 mM ammonium acetate in ESI(−). Conversely, in ESI(+) 5 mM ammonium formate demonstrated superior coverage for diacylglycerols and triacylglycerols.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":null,"pages":null},"PeriodicalIF":2.8000,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1570023224001971/pdfft?md5=6717b93a2a88787f28adc4a713820064&pid=1-s2.0-S1570023224001971-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Improving the intestinal lipidome coverage in a gnotobiotic mouse model using UHPLC-MS-based approach through optimization of mobile phase modifiers and column selection\",\"authors\":\"Habiba Selmi , Alesia Walker , Laurent Debarbieux , Philippe Schmitt-Kopplin\",\"doi\":\"10.1016/j.jchromb.2024.124188\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Lipidomics is focusing on the screening of lipid species in complex mixtures using mass spectrometry-based approaches. 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引用次数: 0
摘要
脂质组学侧重于利用基于质谱的方法筛选复杂混合物中的脂质种类。在这项工作中,我们旨在通过在五种反相色谱柱上测试八种流动相条件,提高寡小鼠-微生物群(OMM12)定植小鼠模型中肠脂质体的覆盖率。我们选择的流动相改良剂包括两种铵盐、两种浓度以及各自添加 0.1% 的酸。我们比较了两种采用混合表面技术的色谱柱、两种采用乙烯桥接混合技术的色谱柱和一种采用核壳颗粒的色谱柱。在所有色谱柱技术中,使用ESI(+)中的甲酸铵或醋酸铵或ESI(-)中的醋酸铵,标准物质和肠道脂质体的性能最佳。值得注意的是,浓度为 5 mM 的铵盐在两种模式下均可获得最佳结果,而添加酸对脂质离子化效率的影响微乎其微。与其他技术相比,HST BEH C18 色谱柱改善了峰宽和尾随因子参数。在 ESI(-)模式下使用 5 mM 乙酸铵时,结肠和回肠中的脂质含量最高,包括神经酰胺、磷脂酰乙醇胺和磷脂酰胆碱。相反,在 ESI(+)条件下,5 mM 甲酸铵对二酰甘油和三酰甘油的覆盖率更高。
Improving the intestinal lipidome coverage in a gnotobiotic mouse model using UHPLC-MS-based approach through optimization of mobile phase modifiers and column selection
Lipidomics is focusing on the screening of lipid species in complex mixtures using mass spectrometry-based approaches. In this work, we aim to enhance the intestinal lipidome coverage within the Oligo-Mouse-Microbiota (OMM12) colonized mouse model by testing eight mobile phase conditions on five reversed-phase columns. Our selected mobile phase modifiers included two ammonium salts, two concentrations, and the addition of respective acids at 0.1 %. We compared two columns with hybrid surface technology, two with ethylene bridged hybrid technology and one with core–shell particles. Best performance was attained for standards and intestinal lipidome, using either ammonium formate or acetate in ESI(+) or ammonium acetate in ESI(−) for all column technologies. Notably, a concentration of 5 mM ammonium salt showed optimal results for both modes, while the addition of acids had a negligible effect on lipid ionization efficiency. The HST BEH C18 column improved peak width and tailing factor parameters compared to other technologies. We achieved the highest lipid count in colon and ileum content, including ceramides, phosphatidylethanolamines and phosphatidylcholines, when using 5 mM ammonium acetate in ESI(−). Conversely, in ESI(+) 5 mM ammonium formate demonstrated superior coverage for diacylglycerols and triacylglycerols.
期刊介绍:
The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis.
Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches.
Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.