Orbitrap Ascend MultiOmics Tribrid mass spectrometer

Importance of the Topic

Multiomics analysis integrates proteomics, metabolomics, lipidomics and glycomics to capture complex biological systems and accelerate discoveries in basic research, drug development and clinical applications.

Study Objectives and Overview

This whitepaper presents innovations in the Orbitrap Ascend MultiOmics Tribrid mass spectrometer designed to scale up multiomics workflows by improving throughput, sensitivity and flexibility. The focus spans multiplexed quantitative proteomics, targeted and untargeted metabolomics, lipidomics, glycoproteomics and glycomics.

Methodology and Instrumentation

The platform leverages a dual ion-routing multipole design that enables parallel HCD fragmentation at MS2 and MS3 stages, coupled with an Ultra-High-Field Orbitrap analyzer delivering >480,000 resolution at up to 45 Hz acquisition. Optional modules include FAIMS Pro Duo, UVPD, ETD/EThcD, PTCR and Native MS modes. Integration with Vanquish Neo UHPLC and μPAC Neo columns, plus an Auto-Ready ion source, enables automated, remote calibration and robust ion delivery.

Main Results and Discussion

• TMT multiplexing with SPS MS3 and Real-Time Search improved quantitative ratio accuracy and increased peptide identifications while reducing fractionation by ~30%.
• DIA workflows achieved coefficients of variance below 10% across sample loads from 50 pg to 500 ng, enabling high-depth, label-free quantitation.
• ETD and EThcD fragmentation raised phosphopeptide and glycopeptide identifications by up to 25% and improved site localization of labile PTMs.
• AcquireX software doubled lipid species coverage through iterative inclusion/exclusion DDA strategies and facilitated double-bond localization with UVPD.
• The SQUAD workflow provided single-injection targeted quantitation and untargeted discovery with femtomole limits of quantitation and six orders of dynamic range.

Benefits and Practical Applications

• Enhanced throughput and reduced experimental costs in quantitative proteomics.
• Comprehensive characterization of labile post-translational modifications and complex lipids.
• Single-analysis workflows for combined targeted and untargeted metabolomics, preserving limited samples.
• Flexibility to analyze sample loads from single cells to bulk preparations without hardware changes.
• Automated maintenance routines for consistent performance, data quality and precise quantitation.

Future Trends and Potential Applications

Next-generation multiomics will leverage AI-driven acquisition strategies, expanded multiplexing chemistries and real-time data analytics. Advances in ion mobility separation, UVPD fragmentation and native MS promise deeper structural insights into isomeric metabolites, lipids and protein complexes.

Conclusion

The Orbitrap Ascend MultiOmics Tribrid mass spectrometer offers a unified, high-performance solution for scaling multiomics research. Its innovative architecture, advanced fragmentation capabilities and automated features deliver unprecedented throughput, sensitivity and versatility, empowering researchers to uncover new biological insights.

Used Instrumentation

• Orbitrap Ascend MultiOmics Tribrid mass spectrometer
• FAIMS Pro Duo interface
• Vanquish Neo UHPLC system with Accucore C30 and μPAC Neo columns
• Dual ion-routing multipoles for parallel HCD at MS2/MS3
• Ultra-High-Field Orbitrap analyzer (>480,000 resolution, 45 Hz)
• Optional UVPD, ETD/EThcD, PTCR and Native MS modules
• Auto-Ready ion source for automated, remote calibration

References

No formal literature references were provided in the source document.