Orbitrap Astral mass spectrometer - Rethink what is possible (2023 year in review)

Importance of the Topic

Mass spectrometry underpins modern analytical chemistry, providing precise identification and quantitation of biomolecules. The advent of combined analyzers like quadrupole, Orbitrap, and Astral enhances sensitivity, resolution, and dynamic range, critical for applications ranging from single-cell omics to plasma proteomics and industrial quality control.

Objectives and Overview of the Study

This body of work reviews the performance of the Thermo Scientific Orbitrap Astral mass spectrometer, focusing on its high-resolution accurate mass (HRAM) capabilities, throughput, and applicability across proteomics, phosphoproteomics, metabolomics, and single-cell analyses. It consolidates results from peer-reviewed studies, technical notes, application white papers, and preprints to evaluate depth of coverage, quantitation precision, and flexibility of experimental workflows.

Methodology and Instrumentation

• Hybrid analyzer design integrating a high-resolution quadrupole, Orbitrap mass analyzer, and Astral mass analyzer.
• Data-independent acquisition (DIA) workflows with narrow-window isolation to maximize peptide identification and quantitation.
• Automated sample preparation via AccelerOme platform for reproducible low-input analyses.
• Complementary front-end solutions: Vanquish Neo UHPLC, μPAC Neo columns, FAIMS Pro Duo interface.
• Software processing through Proteome Discoverer powered by Ardia Platform for streamlined data analysis.

Main Results and Discussion

• Proteomics studies demonstrated identification of over 5,000 proteins per cell in single-cell workflows and deep coverage in bulk samples.
• DIA performance achieved rapid acquisition of HRAM spectra with high sensitivity and dynamic range, reducing cycle times and improving throughput.
• Phosphoproteomics workflows enabled one-hour analyses with robust enrichment strategies, maintaining high site localization confidence.
• Plasma proteomics gained quantitative precision through nanoparticle protein coronas, enabling large cohort studies.
• Metabolomics SQUAD approach combined targeted and untargeted analyses in a single LC-MS run, enhancing discovery potential.

Benefits and Practical Applications

• Unprecedented depth and precision in quantitative proteomics across scales from single cells to tissues and biofluids.
• Streamlined high-throughput workflows for large-cohort biomarker discovery and QA/QC laboratories.
• Enhanced dynamic range and sensitivity supports low-abundance analyte detection in complex matrices.
• Flexible analyzers and software enable simultaneous targeted and untargeted approaches, reducing total analysis time.

Future Trends and Potential Applications

Advancements in multiplexed acquisition, ion mobility integration, and machine learning algorithms for data acquisition and processing are expected to further increase throughput and depth. Emerging applications include immunopeptidomics, clinical proteogenomics, and real-time process monitoring in biomanufacturing.

Conclusion

The Orbitrap Astral mass spectrometer represents a significant leap in mass analyzer technology, offering unmatched HRAM performance, sensitivity, and flexibility. Its integration into diverse workflows empowers researchers and industrial labs to achieve deeper insights, higher throughput, and robust quantitation across proteomics and metabolomics applications.

References

Heil LR, Damoc E, Arrey TN, et al. Evaluating the Performance of the Astral Mass Analyzer for Quantitative Proteomics Using Data-Independent Acquisition. J Proteome Res. 2023;22(10):3290–3300.
Stewart HI, Grinfeld D, Giannakopulos A, et al. Parallelized Acquisition of Orbitrap and Astral Analyzers Enables High-Throughput Quantitative Analysis. Anal Chem. 2023;95(42):15656–15664.
Grinfeld D, Stewart H, Balschun W, et al. Multi-reflection Astral Mass Spectrometer with Isochronous Drift in Elongated Ion Mirrors. Sci Dir. 1060;2024:169017.
Lancaster NM, Sinitcyn P, Forny P, et al. One-Hour Phosphoproteome Analysis with the Orbitrap Astral Mass Spectrometer. bioRxiv. 2023.11.21.568149.
Petrosius V, Arrey TN, Uresin N, et al. Evaluating the Capabilities of the Astral Mass Analyzer for Single-Cell Proteomics. bioRxiv. 2023.06.06.543943.