MSACL 2026 Abstract

A Novel Tribrid LC-MS Workflow for Concurrent Quantification and Untargeted Profiling in Human Plasma for Evaluating Kidney Transplant Outcomes

Antonis Myridakis (1), Alexandra Parada-Torres (1), Sachin Dubei (1), Elliot Murphy (1), Meixuan Li (1), Romina Pacheco-Tapia (2), Inés Castro-Dionicio (2), Francesc Puig-Castellví (2), Olena Potapenko (3), Hendrik Bartolomaeus (4), Johannes Holle (5), Nicola Wilck (3) and Marc-Emmanuel Dumas (1,2*)
(1) Imperial College London, United Kingdom. (2) University of Lille; INSERM-Institut Pasteur de Lille, France. (3) Charité – Universitätsmedizin Berlin, Germany (4) University Hospital Würzburg, Germany (5) University Hospital Tübingen, Germany

Antonis Myridakis, PhD (Presenter) Imperial College London, United Kingdom

Presenter Bio: Antonis is a postdoctoral research associate at the Division of Systems Medicine. He is currently working on the EU-funded IMMEDIATE project developing and utilising metabolomics to identify individual biomarkers of risk and resilience to chronic inflammation, focusing on the interactions within the diet–microbiome–metabolite–immune axis.

His interdisciplinary expertise spans analytical chemistry, biochemistry, exposure science, mass spectrometry and chemometrics, contributing to critical research in public health and environmental toxicology.

His research has been highly cited (h-index 31 / citations >7300) and published in high-impact journals such as Nature, Nature Medicine, and Gut.

Relevant Financial Disclosures (within past 24 months, reported on Apr 21, 2026) No relevant financial relationship(s) to disclose.

INTRODUCTION:
Advancing metabolomics method development requires bridging the gap between broad chemical coverage and precise measurement. While comprehensive metabolomics provides critical systems-level insights into dynamic biochemical changes, traditional analytical methods often force a compromise between untargeted profiling (discovery/hypothesis generation) and targeted quantification (validation/hypothesis confirmation) [1, 2]. To overcome this fundamental limitation, we report a novel workflow based on a SQUAD (Simultaneous Quantification And Discovery) approach utilizing the capabilities of a tribrid (quadrupole/ion trap/orbitrap) instrument [3, 4]. This integrated method enables the concurrent targeted quantification and global profiling of the human plasma metabolome within a single analytical platform, which we demonstrate through a clinical proof-of-concept application for detecting subclinical graft dysfunction in kidney transplantation.

OBJECTIVE(S):
The primary objective of this study is to develop, optimize, and validate an integrated analytical platform capable of simultaneous targeted quantification and broad untargeted profiling, maximizing data richness while conserving sample volume and resources.

METHODS:
The method employs a four-injection high-performance liquid chromatography–mass spectrometry (LC–MS) sequence combining reversed-phase (RP) and hydrophilic interaction (HILIC) separations in positive and negative electrospray ionisation modes. Using a Thermo IQ-X Tribrid mass spectrometer, we quantitatively and sensitively target 150 known metabolites via the linear ion trap with calibration curves while simultaneously acquiring high-resolution full-scan (MS¹) and data-dependent MS/MS spectra for untargeted analysis via the orbitrap mass analyser. Full details about sequence structure and appropriate measures to ensure high quality data for both untargeted and targeted acquisitions are presented.

RESULTS:
All 150 metabolites exhibit linear responses (typically R² > 0.99) across wide concentration ranges with low limits of detection (low/sub ng/mL), enabled by the incorporation of native standards and selected stable-isotope internal standards. The method is validated for precision, accuracy, and matrix effects following standard guidelines. To demonstrate clinical utility, this analytical pipeline is currently being applied to a kidney transplant patient (KTX) cohort as a proof-of-concept to capture dynamic biochemical changes and subclinical graft dysfunction.

CONCLUSION:
This integrated analytical platform overcomes the traditional bottleneck of running separate targeted and untargeted assays. By providing sensitive absolute quantification alongside comprehensive metabolomic profiling, SQUAD streamlines workflows and is exceptionally well-suited for large-scale epidemiological and exposome studies requiring both robust biomarker measurement and novel feature discovery.

REFERENCES:
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