Thermo Scientific™ Orbitrap Exploris™ 240 MS - Versatility–one system with leading performance for proteomics and metabolomics

Importance of Metabolomics Methodology

Metabolomics plays a crucial role in life science research by enabling comprehensive profiling of small molecules in biological systems. Accurate identification and quantitation of metabolites support biomarker discovery, disease diagnostics, and understanding of cellular pathways. A versatile, high‐performance mass spectrometer maximizes data quality across targeted, semi‐targeted, untargeted, and stable isotope labeling workflows, streamlining research and increasing confidence in biological interpretations.

Objectives and Overview of the Study

This proof‐of‐performance study evaluates the Thermo Scientific™ Orbitrap Exploris™ 240 mass spectrometer for broad‐spectrum metabolomics applications. Key goals include demonstrating sub‐ppm mass accuracy, high resolution up to 240,000, extended low‐mass detection, and intelligent data acquisition that together improve compound identification and workflow efficiency.

Methodology and Instrumentation

Sample Sets and Preparation

• Neat standard mixtures of 25 amino acids and vitamins
• Cambridge Isotope Laboratories E. coli cell extract
• NIST SRM1950 human plasma

Instrumentation

• Thermo Scientific Vanquish™ UHPLC system with Hypersil GOLD™ column (15 cm × 2.1 mm, 1.9 µm)
• OptaMax™ NG ion source with EASY-IC internal calibration
• Orbitrap Exploris 240 mass spectrometer
• AcquireX™ intelligent data acquisition workflow
• TraceFinder™ and Compound Discoverer™ software

Chromatography

• Gradient elution from 0% to 98% methanol with 0.1% formic acid over 13 min
• Flow rate: 0.3 mL/min

Main Results and Discussion

Sub‐ppm Mass Accuracy and High Resolution

• Repeated analyses of standard mixtures over five days on three instruments achieved mass errors below 1 ppm in both positive and negative mode.
• Resolution up to 240,000 resolved fine isotope patterns and heteroatom contributions in unknown compounds, expediting elemental formula prediction.

Extended Low‐Mass Range

• Detection down to m/z 40 recovered additional fragment ions, enabling differentiation of isomeric metabolites (e.g., 2-aminobutyric acid vs. dimethylglycine).

Intelligent AcquireX Acquisition

• Automated blank exclusion and iterative sampling increased unique MS/MS identifications in human plasma compared to traditional DDA.
• Background ions, adducts, and redundancies are filtered out on‐the‐fly, maximizing biologically relevant spectra.

Benefits and Practical Applications

The Orbitrap Exploris 240 MS integrates proteomics and metabolomics on a single platform, delivering reproducible high‐quality data. Sub‐ppm mass accuracy and fine isotope resolution support confident compound annotation. Extended low‐mass coverage and AcquireX intelligence enhance detection of low‐abundance metabolites and isomers, improving throughput and depth in biomarker discovery and QA/QC workflows.

Future Trends and Opportunities

Emerging developments include further automation of data processing, integration with ion mobility separations, and application of machine‐learning algorithms for annotation and pathway mapping. Improvements in acquisition strategies will continue to expand metabolome coverage, enabling large‐scale clinical and environmental studies.

Conclusion

The Orbitrap Exploris 240 mass spectrometer redefines metabolomics performance by combining ultra‐high resolution, sub‐ppm accuracy, extended low‐mass detection, and advanced AcquireX acquisition. This platform empowers researchers with rapid, reliable metabolite profiling for diverse life science applications.