Thermo Scientific Orbitrap Fusion Lumos Tribrid Mass Spectrometer

Importance of the topic

Mass spectrometry is a cornerstone of analytical chemistry, enabling precise measurement of molecular structure, abundance and modifications. The evolution of hybrid instruments offers enhanced sensitivity, mass accuracy and dynamic range, essential for proteomics, metabolomics, lipidomics, pharmaceutical analysis and biomarker discovery.

Objectives and study overview

This article introduces the Thermo Scientific Orbitrap Fusion Lumos Tribrid mass spectrometer and demonstrates its performance across diverse applications. Key goals include multiplexed targeted proteomics, data independent acquisition for large-scale quantitation, top-down analysis of intact proteins, comprehensive glycopeptide characterization, lipid profiling with polarity switching and small molecule identification at femtogram levels.

Methodology and instrumentation

The Tribrid architecture integrates:

• High capacity transfer tube and electrodynamic ion funnel for efficient ion transmission
• Advanced Active Ion Beam Guide to reject neutrals
• Dual-pressure linear ion trap for fast MSn fragmentation (CID, HCD, ETD, EThcD)
• Quadrupole for high-resolution precursor isolation
• Ion routing multipole enabling parallel fragmentation and analysis
• Ultra-high field Orbitrap analyzer offering >500,000 FWHM and scan rates up to 30 Hz
• Advanced vacuum technology for optimal transmission
• Integrated intuitive method editor with preconfigured templates

Main results and discussion

1. Multiplexed targeted proteomics: The TMT SPS tMS3 workflow quantifies ten samples simultaneously with improved accuracy even at low peptide levels. Monitoring 131 peptides across 69 proteins in NCI-60 cell lines (180 samples in 48 hours) showed reproducible fold changes and drug-response correlations.
2. Data independent acquisition: Step-wise DIA with 15 amu windows delivered high resolution MS2 data for a HeLa digest, identifying and quantifying over 92% of protein groups and 85% of peptides with CV <7% using a spectral library approach.
3. Parallel reaction monitoring: PRM assays achieved attomole limits of quantitation (1 amol) for spiked peptides in complex matrices, with CVs <11% at low levels.
4. Top-down protein isoform analysis: Advanced Vacuum Technology and improved quadrupole isolation resolved histone H3 methylation isoforms and delivered high sequence coverage via ETD HD, enabling detection of disease-relevant proteoforms.
5. Intact monoclonal antibody characterization: Baseline resolution of mAb glycoforms, isotopic resolution of heavy and light chains and combined CID, HCD and ETD HD fragmentation yielded up to 91% sequence coverage for light chains and 62% for heavy chains.
6. Glycoproteomics: HCD product-dependent EThcD/CID workflows provided comprehensive glycopeptide identification, enhanced sequence coverage and precise glycosylation site localization in a single analysis.
7. Lipid profiling: Fast polarity switching and intelligent HCD/CID acquisition enabled simultaneous identification of phosphatidylcholines and co-eluting triacylglycerol isomers in one LC-MS run, improving throughput and confidence.
8. Small molecule analysis: High resolution MS2 of norfloxacin fine isotopic structure unambiguously confirmed elemental composition. Quantitation of Irganox 1035 in food simulant achieved a LOQ of 100 fg with five orders of linear dynamic range and CVs <10%.

Benefits and practical applications

The Orbitrap Fusion Lumos MS delivers superior sensitivity, selectivity and versatility, reducing analysis time, improving quantitative accuracy at low concentrations and enabling detailed characterization of complex biomolecules. Laboratories benefit from streamlined workflows for high-throughput proteomics, QA/QC in pharmaceutical development, environmental screening and translational research.

Future trends and opportunities

Emerging directions include integration of data-driven acquisition strategies, artificial intelligence for real-time method optimization, microflow and single-cell platforms, novel fragmentation chemistries and expanding multiplexing capabilities. Advances in informatics and cloud-based analysis will further accelerate biological discovery.

Conclusion

The Thermo Scientific Orbitrap Fusion Lumos Tribrid mass spectrometer represents a significant advance in mass spectrometry technology. Its combination of high resolution, versatile fragmentation modes and intelligent acquisition tools addresses critical analytical challenges and opens new horizons across life sciences and small molecule research.

References

• Everley et al. WP 663, ASMS 2016
• Zheng Y et al. Mol. Cell Proteomics 2016, 15, 776–90