Increasing plant metabolome coverage using Orbitrap enhanced Dynamic Range (eDR) scan mode on an Orbitrap Excedion Pro mass spectrometer

Importance of Enhanced Dynamic Range in Plant Metabolomics

Effective profiling of plant metabolites is crucial for understanding biochemical pathways, quality control in herbal products, and discovery of bioactive compounds. Enhancing signal‐to‐noise for low‐abundance molecules improves coverage and identification confidence, directly impacting research in phytochemistry and agricultural science.

Study Objectives and Overview

This study evaluates how the Orbitrap Excedion Pro mass spectrometer’s enhanced Dynamic Range (eDR) scan mode expands metabolome coverage at the MS1 level and improves fragment‐based annotation using a data‐dependent AcquireX Deep Scan workflow at the MS2 level. Chamomile tea extracts serve as a complex model sample to benchmark performance against conventional full-scan acquisition.

Methodology

Chromatographic separation was performed on a Thermo Scientific Hypersil GOLD VANQUISH C18 UHPLC column with water/methanol (0.1% formic acid) gradient at 0.3 mL/min. MS1 profiling used 120 000 resolution with eDR on/off. MS2 data acquisition employed a Thermo Scientific AcquireX Deep Scan workflow (10 injections) at 15 000 resolution to collect fragmentation spectra for enhanced annotation.

Instrumental Configuration

The key innovation lies in the Orbitrap Excedion Pro’s intelligent MS1 multiplexing, dividing 67–1000 m/z into two alternating subscans with defined windows. Quadrupole isolation pairs with optimized injection timing to maximize signal for low- and medium-intensity ions during a single Orbitrap injection.

Main Results and Discussion

Comparing full-scan data, eDR ON yielded a 2.6-fold increase in detected compounds (CV ≤20%). Signal‐to‐noise ratios for medium and low-abundance features improved substantially, enabling detection of ions absent in legacy scans. MS2 annotation using mzCloud library revealed a 50% rise in identified compounds, with more than double the number of fragmentation spectra recorded. These gains translated into deeper metabolome coverage and higher-confidence formula assignments.

Practical Benefits and Applications

• Broadens detection range to include trace-level metabolites in complex botanical extracts.
• Enhances reproducibility (reduced CVs) for quality control in industrial and research labs.
• Facilitates more comprehensive untargeted metabolomics studies through improved annotation rates.

Future Trends and Potential Uses

Adoption of eDR strategies is expected to accelerate in plant metabolomics, environmental analysis, and clinical research. Combining eDR with emerging acquisition methods (e.g., parallel reaction monitoring, ion mobility) may further enhance depth and throughput. Software advances in automated background subtraction and spectral stitching will complement hardware improvements.

Conclusion

The Orbitrap Excedion Pro’s eDR mode significantly outperforms conventional full-scan acquisition for untargeted plant metabolomics. By boosting compound discovery and fragment-based annotation, eDR enables more comprehensive studies of complex samples without sacrificing quantitation precision.

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