Stettin Daniel, Poulin Remington X, Pohnert Georg
Institute for Inorganic and Analytical Chemistry, Bioorganic Analytics, Friedrich Schiller University Jena, 07743 Jena, Germany.
Metabolites. 2020 Apr 4;10(4):143. doi: 10.3390/metabo10040143.
The development of improved mass spectrometers and supporting computational tools is expected to enable the rapid annotation of whole metabolomes. Essential for the progress is the identification of strengths and weaknesses of novel instrumentation in direct comparison to previous instruments. Orbitrap liquid chromatography (LC)-mass spectrometry (MS) technology is now widely in use, while Orbitrap gas chromatography (GC)-MS introduced in 2015 has remained fairly unexplored in its potential for metabolomics research. This study aims to evaluate the additional knowledge gained in a metabolomics experiment when using the high-resolution Orbitrap GC-MS in comparison to a commonly used unit-mass resolution single-quadrupole GC-MS. Samples from an osmotic stress treatment of a non-model organism, the microalga , were investigated using comparative metabolomics with low- and high-resolution methods. Resulting datasets were compared on a statistical level and on the level of individual compound annotation. Both MS approaches resulted in successful classification of stressed vs. non-stressed microalgae but did so using different sets of significantly dysregulated metabolites. High-resolution data only slightly improved conventional library matching but enabled the correct annotation of an unknown. While computational support that utilizes high-resolution GC-MS data is still underdeveloped, clear benefits in terms of sensitivity, metabolic coverage, and support in structure elucidation of the Orbitrap GC-MS technology for metabolomics studies are shown here.
改进型质谱仪及相关计算工具的发展有望实现对整个代谢组的快速注释。与之前的仪器直接比较，识别新型仪器的优缺点对于这一进展至关重要。轨道阱液相色谱（LC）-质谱（MS）技术现已广泛应用，而2015年推出的轨道阱气相色谱（GC）-MS在代谢组学研究中的潜力仍未得到充分探索。本研究旨在评估与常用的单位质量分辨率单四极杆GC-MS相比，使用高分辨率轨道阱GC-MS进行代谢组学实验时所获得的额外知识。使用低分辨率和高分辨率方法，通过比较代谢组学对非模式生物微藻经渗透胁迫处理后的样品进行了研究。对所得数据集在统计层面和单个化合物注释层面进行了比较。两种质谱方法均成功区分了胁迫和未胁迫的微藻，但使用的是不同的显著失调代谢物集。高分辨率数据仅略微改善了传统的库匹配，但能够正确注释一种未知物。虽然利用高分辨率GC-MS数据的计算支持仍不发达，但本文展示了轨道阱GC-MS技术在代谢组学研究的灵敏度、代谢覆盖范围和结构解析支持方面的明显优势。
