Systems biology approaches increasingly rely on the integration of metabolomics data across multiple biological matrices to capture systemic metabolic variation. However, a major limitation remains the lack of standardized workflows enabling comparable metabolomic profiling across different human biofluids.
High-field Nuclear Magnetic Resonance (NMR) spectroscopy is a cornerstone of metabolomics, but its high cost and the need for specialized infrastructure limit its translation into clinical practice. Recently, benchtop low-field (LF) NMR spectrometers have emerged as a more accessible alternative, that could break down the cost barrier potentially enabling metabolomic analyses in virtually any setting.
In this study, we developed and validated a standardized benchtop NMR workflow for metabolomic profiling of multiple human biofluids, including serum and urine, to enable harmonized metabolic measurements across matrices.
The workflow includes sample preparation protocols, standardized benchtop NMR acquisition parameters, and automated spectral processing. Protocol robustness was validated through cross-platform comparison between low-field (100 MHz) and high-field (600 MHz) NMR spectroscopy.
As a proof-of-concept, the workflow was applied to a pilot cohort including serum and urine samples collected from patients before and after treatment. Multivariate analyses revealed treatment-associated metabolic shifts, demonstrating the potential of the platform for longitudinal metabolic profiling.

Co-authors: Linda Fantato, Valentina Giraldi, Leonardo Tenori