# Enhancing detection of low‑abundance metabolites in proton NMR through band‑selective suppression and presaturation

## TL;DR

This paper introduces a new NMR method to better detect low-abundance metabolites by suppressing strong signals from common compounds like sugars.

## Contribution

A novel 1D presat-1H-ES NMR method combining presaturation and excitation sculpting to enhance low-abundance metabolite detection.

## Key findings

- The method provides 2–4 times signal enhancement for low-abundance metabolites in sugar-rich samples.
- It improves reproducibility and discrimination in multivariate analyses compared to conventional methods.
- The approach is broadly applicable to various systems where dominant metabolites obscure weaker signals.

## Abstract

Metabolomics provides powerful means to analyze metabolite profiles in biological samples, enabling insights into biochemical changes under genetic, environmental, or pathological conditions. Nuclear Magnetic Resonance (NMR) spectroscopy is central to metabolomics, but its utility is often constrained by the strong and overlapping resonances of abundant components, such as sugars in plant‑ and food‑derived materials, which obscure signals of lower‑abundance metabolites. Here, we introduce a modified NMR acquisition method that increases sensitivity and specificity by selectively suppressing dominant signals, while enhancing weaker metabolite signals across the spectrum. The method integrates water presaturation with excitation sculpting (ES), yielding a robust 1D presat‑1H‑ES pulse sequence. Validation on a range of sugar-rich samples demonstrated 2–fourfold signal enhancement for low‑abundance metabolites compared with conventional 1H‑ES. Multivariate analyses show the method improves reproducibility and discrimination, enabling detection and comparison of low‑abundance metabolites not accessible with conventional approaches’. Beyond sugar‑rich systems, the method is broadly applicable to other spectral regions where dominant metabolite classes obscure lower‑concentration compounds, including primary metabolites and structurally diverse natural products. Overall, the 1D presat‑1H‑ES significantly enhances resolution and sensitivity of NMR‑based metabolomics, shortens analysis time, and supports more precise profiling for both fundamental studies and translational applications in metabolomics and natural‑products discovery.

The online version contains supplementary material available at 10.1007/s13659-025-00570-3.

Robust 1D presat‑1H‑ES NMR method integrating presaturation and excitation sculpting to selectively suppress dominant signals.Validated on plant‑derived and food materials, improving sensitivity, reproducibility, and multivariate discrimination.Fast, user‑friendly, high‑throughput solution for metabolic profiling with broad applications in food quality, nutrition, and natural‑product research.

Robust 1D presat‑1H‑ES NMR method integrating presaturation and excitation sculpting to selectively suppress dominant signals.

Validated on plant‑derived and food materials, improving sensitivity, reproducibility, and multivariate discrimination.

Fast, user‑friendly, high‑throughput solution for metabolic profiling with broad applications in food quality, nutrition, and natural‑product research.

The online version contains supplementary material available at 10.1007/s13659-025-00570-3.