Univ Rouen Normandie, INSA Rouen Normandie, CNRS, Normandie Univ, COBRA UMR 6014, INC3M FR 3038, F-76000, Rouen, France.
Université de Lyon, Institut des Sciences Analytiques, UMR 5280 CNRS, 5 rue de la Doua, 69100, Villeurbanne, France.
Anal Chim Acta. 2024 May 22;1304:342535. doi: 10.1016/j.aca.2024.342535. Epub 2024 Mar 26.
The implementation of ion mobility spectrometry (IMS) in liquid chromatography-high-resolution mass spectrometry (LC-HRMS) workflows has become a valuable tool for improving compound annotation in metabolomics analyses by increasing peak capacity and by adding a new molecular descriptor, the collision cross section (CCS). Although some studies reported high repeatability and reproducibility of CCS determination and only few studies reported good interplatform agreement for small molecules, standardized protocols are still missing due to the lack of reference CCS values and reference materials. We present a comparison of CCS values of approximatively one hundred lipid species either commercially available or extracted from human plasma. We used three different commercial ion mobility technologies from different laboratories, drift tube IMS (DTIMS), travelling wave IMS (TWIMS) and trapped IMS (TIMS), to evaluate both instrument repeatability and interlaboratory reproducibility. We showed that CCS discrepancies of 0.3% (average) could occur depending on the data processing software tools. Moreover, eleven CCS calibrants were evaluated yielding mean RSD below 2% for eight calibrants, ESI Low concentration tuning mix (Tune Mix) showing the lowest RSD (< 0.5%) in both ion modes. Tune Mix calibrated CCS from the three different IMS instruments proved to be well correlated and highly reproducible (R > 0.995 and mean RSD ≤ 1%). More than 90% of the lipid CCS had deviations of less than 1%, demonstrating high comparability between techniques, and the possibility to use the CCS as molecular descriptor. We highlighted the need of standardized procedures for calibration, data acquisition, and data processing. This work demonstrates that using harmonized analytical conditions are required for interplatform reproducibility for CCS determination of human plasma lipids.
在液相色谱-高分辨质谱(LC-HRMS)工作流程中实施离子淌度谱(IMS)已成为通过增加峰容量并增加新的分子描述符,碰撞截面(CCS),来改善代谢组学分析中化合物注释的有效工具。尽管一些研究报告称 CCS 测定具有很高的重复性和重现性,并且只有少数研究报告称小分子的跨平台一致性良好,但由于缺乏参考 CCS 值和参考物质,因此仍然缺乏标准化的协议。我们比较了大约一百种商业上可获得的或从人血浆中提取的脂质物种的 CCS 值。我们使用了来自三个不同实验室的三种不同的商业离子淌度技术,即漂移管 IMS(DTIMS)、行波 IMS(TWIMS)和俘获 IMS(TIMS),以评估仪器重复性和实验室间重现性。结果表明,CCS 值可能会因数据处理软件工具而异,差异为 0.3%(平均值)。此外,评估了十一种 CCS 校准标品,对于八种校准标品,ESI 低浓度调谐混合物(Tune Mix)在两种离子模式下的 RSD 均低于 2%。由三种不同的 IMS 仪器校准的 Tune Mix CCS 证明具有很好的相关性和高度重现性(R>0.995,平均 RSD≤1%)。超过 90%的脂质 CCS 的偏差小于 1%,表明技术之间具有高度的可比性,并且可以将 CCS 用作分子描述符。我们强调了需要标准化的程序,包括校准、数据采集和数据处理。这项工作表明,使用协调一致的分析条件对于人血浆脂质 CCS 测定的跨平台重现性是必需的。