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High-Throughput Equine Plasma Profiling Using Laser-Assisted Rapid Evaporative Ionisation Mass Spectrometry.
Abstract: Remnants of erythrocyte removal from blood constitute therapeutic plasma-a water-based solution containing mainly platelets, extracellular vesicles (EVs), protein and lipid complexes and ions. Plasma-based therapies, including their use to accelerate wound healing in human and animal patients, are increasingly gaining prominence in regenerative medicine; however, methods for the optimal preparation of individual plasma are still under development. Rapid and accurate analysis of plasma composition is essential for the development of therapeutic applications and for ensuring the quality of plasma products before patient administration. This study of equine plasma presents a novel approach that utilises a newly developed automated well-plate sampling system using laser-assisted rapid evaporative ionisation mass spectrometry (LA-REIMS) for rapid plasma profiling under varying sample preparation conditions. This platform provided comprehensive lipidomic and metabolic analyses that gave insights into the molecular composition of plasma within seconds and without extensive sample preparation. The results indicated that specific phosphatidylcholines (PC(34:2), PC(36:2), PC(36:3)) and fatty acids were sensitive to centrifugation parameters, suggesting their potential as indicators of plasma stability and quality. The results were compared with the platelet and EV content of the plasma. The number density (n) of platelets and cellular fragments (P2 particles) was determined by flow cytometry (FCM), whereas EVs were characterised by interferometric light microscopy (ILM). The samples were imaged by scanning electron microscopy (SEM). A clear correlation was observed between n of P2 particles (but not of EVs) and lipid profiles, suggesting platelets' critical role in plasma composition. These findings emphasise the potential of the LA-REIMS platform for high-throughput, rapid plasma analysis, quality control and diagnostics. The system's ability to quickly assess plasma quality highlights its practical use in validating plasma prior to therapeutic applications.
© 2026 The Author(s). Journal of Mass Spectrometry published by John Wiley & Sons Ltd.
Publication Date: 2026-07-23 PubMed ID: 42489316PubMed Central: PMC13393337DOI: 10.1002/jms.70077Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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Overview
- This research presents a new rapid method for analyzing horse plasma composition using laser-assisted rapid evaporative ionization mass spectrometry (LA-REIMS).
- The study aims to improve the preparation and quality control of plasma used in regenerative therapies by providing fast and detailed molecular profiles.
Introduction and Background
- Plasma is a water-based solution derived after removing red blood cells from blood, containing platelets, extracellular vesicles (EVs), proteins, lipids, and ions.
- Plasma-based therapies are increasingly important in regenerative medicine for accelerating wound healing in humans and animals.
- Optimizing plasma preparation methods is a current challenge that requires rapid and accurate plasma composition profiling.
- Traditional analytical methods for assessing plasma quality can be time-consuming and require extensive sample preparation.
Methodology
- The study uses an innovative automated well-plate system combined with laser-assisted rapid evaporative ionization mass spectrometry (LA-REIMS) for rapid profiling.
- This platform allows for comprehensive lipidomic and metabolic analysis directly from plasma samples within seconds without extensive sample treatment.
- Different sample preparation conditions, mainly centrifugation parameters, are tested to observe their effect on plasma molecular composition.
- Flow cytometry (FCM) is employed to quantify the number density of platelets and cellular fragments (referred to as P2 particles).
- Extracellular vesicles (EVs) are characterized using interferometric light microscopy (ILM).
- Scanning electron microscopy (SEM) images are used to visualize the plasma samples for structural insights.
Key Findings
- Specific lipid molecules, including phosphatidylcholines PC(34:2), PC(36:2), PC(36:3), and fatty acids, are sensitive to changes in centrifugation parameters.
- These lipid molecules act as potential indicators of plasma stability and quality, aiding in assessment of plasma preparations.
- A clear correlation exists between the number density of P2 particles (platelets and fragments) and the lipidomic profiles measured by LA-REIMS.
- No significant correlation is found between EV number density and the lipid profile, highlighting that platelets have a dominant influence on plasma composition.
- SEM imaging supports the observations by visually confirming the presence and condition of cellular fragments and platelets in plasma samples.
Implications and Applications
- The LA-REIMS platform provides a high-throughput, rapid, and minimally invasive method for plasma profiling.
- This approach facilitates quick, accurate evaluation of plasma molecular composition, which is crucial for quality control in therapeutic settings.
- The ability to rapidly assess plasma quality helps ensure that plasma products used in regenerative medicine meet standards for safety and efficacy before patient administration.
- The methodology could be applied broadly in veterinary and human plasma therapies, potentially improving outcomes by tailoring plasma preparation to molecular quality indicators.
- Overall, this research demonstrates the utility of LA-REIMS as a powerful tool in plasma diagnostics, quality control, and therapeutic plasma product development.
Cite This Article
APA
Molnár A, Arko M, Troha K, Marton A, Tauber B, Iglič A, Balogh GT, Balog J, Hočevar M, Kos VK, Battelino S, Kralj-Iglič V, Schlosser G.
(2026).
High-Throughput Equine Plasma Profiling Using Laser-Assisted Rapid Evaporative Ionisation Mass Spectrometry.
J Mass Spectrom, 61(8), e70077.
https://doi.org/10.1002/jms.70077 Publication
Researcher Affiliations
- MTA-ELTE Lendület (Momentum) Ion Mobility Mass Spectrometry Research Group, Faculty of Science, Institute of Chemistry, ELTE Eötvös Loránd University, Budapest, Hungary.
- Hevesy György PhD School of Chemistry, ELTE Eötvös Loránd University, Budapest, Hungary.
- Laboratory of Clinical Biophysics, Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia.
- Department of Otorhinolaryngology and Cervicofacial Surgery, University Medical Centre Ljubljana, Ljubljana, Slovenia.
- Department of Chemical and Environmental Process Engineering, Budapest University of Technology and Economics, Budapest, Hungary.
- Ambimass Kft., Budapest, Hungary.
- Qamcom Central Europe, Budapest, Hungary.
- Laboratory of Physics, Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia.
- Department of Chemical and Environmental Process Engineering, Budapest University of Technology and Economics, Budapest, Hungary.
- Department of Pharmaceutical Chemistry, Semmelweis University, Budapest, Hungary.
- Center for Pharmacology and Drug Research & Development, Semmelweis University, Budapest, Hungary.
- Waters Corporation, Cambridge, Massachusetts, USA.
- Institute of Metals and Technology, Ljubljana, Slovenia.
- Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia.
- Department of Otorhinolaryngology and Cervicofacial Surgery, University Medical Centre Ljubljana, Ljubljana, Slovenia.
- Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
- Laboratory of Clinical Biophysics, Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia.
- MTA-ELTE Lendület (Momentum) Ion Mobility Mass Spectrometry Research Group, Faculty of Science, Institute of Chemistry, ELTE Eötvös Loránd University, Budapest, Hungary.
MeSH Terms
- Animals
- Horses / blood
- Blood Platelets / chemistry
- Plasma / chemistry
- Mass Spectrometry / methods
- High-Throughput Screening Assays / methods
- Lipidomics / methods
- Extracellular Vesicles / chemistry
- Lasers
- Phosphatidylcholines / blood
- Lipids / blood
Grant Funding
- SNN 148580 / National Research, Development and Innovation Fund
- 148580 / National Research, Development and Innovation Fund
- P2-0132 / Slovenian Research and Innovation Agency
- P2-0232 / Slovenian Research and Innovation Agency
- P3-0388 / Slovenian Research and Innovation Agency
- P4-0053 / Slovenian Research and Innovation Agency
- J3-60063 / Slovenian Research and Innovation Agency
- Hungarian Academy of Sciences
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