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Dec 23, 2025 | Blogs, Life Science Research, ZenoTOF 7600 system, ZenoTOF 8600 system | 0 comments
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In the field of food chemistry and health, Prof. Nils Helge Schebb and his team at the University of Wuppertal are at the forefront of applying cutting-edge analytical methods to investigate how dietary components affect inflammation and chronic disease. Their work focuses on lipid mediators, particularly oxylipins, and how these molecules can be precisely measured and interpreted using liquid chromatography-tandem mass spectrometry (LC-MS).
Why is LC-MS the gold standard for oxylipin analysis?
Oxylipins are bioactive lipid mediators present in low concentrations and prone to degradation. This makes their analysis particularly challenging. LC-MS provides the sensitivity, specificity, and resolution necessary to quantify these compounds in complex biological matrices accurately. Prof. Schebb’s team has developed and refined robust LC-MS workflows that allow researchers to detect and quantify oxylipins with high precision, enabling deeper insights into how diet modulates inflammation at the molecular level.
How should samples be prepared for reliable oxylipin quantitation?
Sample preparation is a critical step in any LC-MS workflow. Prof. Schebb recommends using solid-phase extraction (SPE) and liquid-liquid extraction (LLE) techniques to effectively isolate oxylipins from biological matrices such as plasma, serum, or tissue. To preserve the integrity of these labile molecules, it is essential to minimize their exposure to light, heat, and oxygen throughout the preparation process. Additionally, incorporating antioxidants and enzyme inhibitors during extraction helps prevent the artificial formation or degradation of oxylipins, ensuring that the results reflect true biological conditions.
Best practice: Careful sample handling and preparation are essential to avoid false positives and ensure reproducibility.
What are the optimal chromatographic conditions for oxylipin separation?
To achieve optimal separation of oxylipins, Prof. Schebb’s team employs reverse-phase liquid chromatography (RPLC) using C18 columns, which are well-suited for lipid analysis. The chromatographic method employs gradient elution with a mixture of water and acetonitrile, both acidified with formic acid, to improve peak shape and retention. Run times are carefully optimized to strike a balance between analytical depth and throughput, allowing for efficient separation of structurally similar oxylipins, an essential factor for accurate quantitation and biological interpretation.
How does tandem mass spectrometry improve detection?
Tandem mass spectrometry (MS/MS), particularly when operated in multiple reaction monitoring (MRM) mode, is a central component of Prof. Schebb’s analytical strategy. This approach enables the selective detection of target oxylipins based on their unique fragmentation patterns. It also offers high sensitivity, allowing for the detection of oxylipins at picomolar concentrations. When paired with stable isotope-labelled internal standards, MS/MS provides the quantitative accuracy needed for reliable lipid mediator profiling.
Key insight: MS/MS in MRM mode is essential for distinguishing oxylipins from isobaric interferences and background noise.
How is data quality ensured in oxylipin analysis?
Prof. Schebb emphasizes the importance of rigorous quality control throughout the analytical pipeline. The use of internal standards helps correct for variability in both extraction and ionization efficiency. Calibration curves are employed to ensure linearity and accuracy across a wide range of concentrations. Additionally, system suitability tests and method validation procedures are conducted regularly to maintain consistent instrument performance and data reliability.
Quantitation of oxylipins, especially in complex matrices, has long been a topic of debate. Recently published guidelines now provide a framework for reporting oxylipin measurements. To ensure compliance, the lower limit of quantitation (LLOQ) for each compound was defined as the lowest standard injection yielding a signal-to-noise ratio (S/N) greater than 5. Crucially, this determination must be based on raw, unsmoothed data. Applying smoothing techniques can artificially reduce noise levels, resulting in inflated and potentially misleading signal-to-noise (S/N) values. This is particularly important in complex matrices, where background noise may be misinterpreted as a true analyte signal.
Pro tip: High-quality data starts with validated methods and ends with robust QC protocols.
What role does chiral separation play in lipid mediator research?
Many oxylipins are chiral molecules, meaning they exist as mirror-image forms (enantiomers) that can have distinct biological activities. Prof. Schebb’s team utilizes chiral high-performance liquid chromatography (HPLC) to separate these enantiomers, enabling a more nuanced understanding of their physiological roles. This approach enables researchers to determine the stereospecific effects of lipid mediators on inflammation and disease, thereby enhancing the biological relevance of lipidomics data.
Why it matters: Without chiral separation, researchers risk overlooking critical differences in bioactivity between enantiomers.
How do these methods advance personalized nutrition and health?
By combining targeted LC-MS workflows, oxylipin metabolomics, and chiral analysis, Prof. Schebb’s research provides a molecular framework for understanding how diet influences health. This integrated approach facilitates the development of personalized dietary recommendations tailored to individual lipid mediator profiles. It also enables the discovery of novel biomarkers for inflammation and chronic disease and offers mechanistic insights into how specific nutrients modulate physiological pathways.
How are SCIEX ZenoTOF systems advancing research on lipid mediators?
To push the boundaries of lipidomics and oxylipin analysis, researchers are increasingly turning to high-resolution mass spectrometry (HRMS) platforms. The SCIEX ZenoTOF 7600 and 8600 systems are at the forefront of this analytical evolution, offering unmatched capabilities for lipid mediator profiling.
What makes the ZenoTOF 7600/8600 systems ideal for oxylipin analysis?
The ZenoTOF series combines ultra-high resolution, sensitivity, and speed, making it ideal for detecting low-abundance, labile lipid mediators such as oxylipins. Key advantages include:
Performance Edge: The ZenoTOF 8600 system offers up to 20 times greater sensitivity in MS/MS mode, enabling confident detection of low-abundance oxylipins that conventional QTOF systems may miss.
How does electron-activated dissociation (EAD) enhance structural characterization?
One of the most powerful features of the ZenoTOF 7600 and 8600 systems is electron-activated dissociation (EAD), a novel fragmentation technique that provides rich structural information without over-fragmenting labile molecules. For lipid mediators, EAD enables the precise localization of double bonds and hydroxyl groups, which are critical for identifying oxylipin isomers. It also preserves labile modifications, such as epoxides and hydroperoxides, that are often lost during traditional collision-induced dissociation (CID). Additionally, EAD supports the differentiation of chiral and regioisomers, allowing for more accurate biological interpretation.
Analytical breakthrough: EAD empowers researchers to go beyond quantitation and into structural elucidation, revealing the subtle differences that define oxylipin function.
How do these technologies support translational research?
By integrating ZenoTOF HRMS with targeted lipidomics workflows, scientists can monitor diet-induced changes in oxylipin profiles with high confidence. These technologies also enable the discovery of novel lipid biomarkers for inflammation, aging, and chronic disease. Most importantly, they help translate molecular insights into personalized nutrition strategies and clinical applications, bridging the gap between analytical chemistry and real-world health outcomes.
Big picture: SCIEX ZenoTOF platforms are not just analytical tools; they’re enablers of discovery in systems biology, nutrition science, and precision medicine.
Find out more about Prof. Nils Helge Schebb and his team’s work analysing oxylipins, or how advanced analytical techniques have the potential to revolutionize their analysis here:
Technical recommendations for analyzing oxylipins by liquid chromatography–mass spectrometry https://www.science.org/doi/10.1126/scisignal.adw1245
Quantitative and qualitative analysis of oxylipins using the ZenoTOF 8600 system
Quantitative analysis of lipid mediators using the ZenoTOF 7600 system
Learn more about EAD: https://sciex.com/technology/electron-activated-dissociation
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