Orbitrap Astral mass spectrometer - Brochure

Significance of the Topic

Mass spectrometry-driven proteomics underpins our understanding of biology and disease by identifying and quantifying proteins in complex samples. However, traditional instruments face trade-offs between throughput, depth, sensitivity, and quantitative precision that limit large-cohort and low-input analyses. The Thermo Scientific Orbitrap Astral mass spectrometer addresses these limitations by combining novel high-speed, high-sensitivity detection with high-resolution analyzers, enabling comprehensive, high-throughput proteomic workflows.

Study Objectives and Overview

This whitepaper introduces the Orbitrap Astral mass spectrometer, detailing how its dual-analyzer architecture and end-to-end Velocity workflow expand experimental capabilities. By integrating an innovative Astral mass analyzer with a proven Orbitrap detector and complementary components, the aim is to demonstrate improvements in throughput, coverage, sensitivity, and quantitative accuracy for diverse proteomic applications.

Methodology and Instrumentation

The instrument architecture comprises three mass analyzers:

• Quadrupole for high-speed precursor selection
• Ultra-high-field Orbitrap for panoramic, high-resolution, high-mass-accuracy full scans
• Astral analyzer for rapid (up to 200 Hz), high-sensitivity, high dynamic range MS2 detection

Key components include:

• Ion source with enhanced transmission and brightness
• Asymmetric ion mirrors and long ion track for resolution ≥80,000
• Ion routing multipole and ion processor for efficient trapping and fragmentation
• High dynamic range detector for low-noise, single-ion sensitivity
• Thermo Scientific Velocity workflow: sample preparation (AccelerOme), chromatography (Vanquish Neo UHPLC, µPAC and EASY-Spray columns), ion mobility (FAIMS Pro Duo), and data processing (Proteome Discoverer with CHIMERYS and Ardia Platform)

Key Findings and Discussion

• Throughput: Achieves >8,000 protein groups in an 8-minute single-shot run (180 samples/day, ~1.4 million protein IDs/day), a 4× speed increase over previous platforms.
• Depth: Identifies ~9,000 proteins in 15 min and ~12,000 in 60 min single injections; enables near-whole-proteome coverage (15,000+ proteins) in 4.5 h using fractionated samples.
• Sensitivity: Detects >5,000 proteins from 250 pg HeLa digest at 80 samples/day; low-input runs show >75% proteins with CV < 10% and data completeness up to 90%.
• Single-cell proteomics: DirectDIA yields >5,300 proteins per 250 pg cell equivalent at 80 runs/day, doubling throughput and coverage versus competing instruments.
• Quantitative precision: LFQ HR-DIA of three-proteome mixes (Human/E. coli/Yeast) identifies ~14,000 proteins with median CV ≤ 10% and accurate ratios across a 1.5–9× dynamic range.
• TMT multiplexing: Resolves TMTpro 18plex reporter ions and peptide fragments simultaneously; quantifies >11,000 proteins at 40 min/sample (36 samples/day), exceeding state-of-the-art throughput and depth.
• Phosphoproteomics: Doubles depth of phosphopeptide coverage versus current Orbitrap MS, reaching ~20,000 Class I sites in 8 min (4× throughput).
• Plasma proteomics: Profiles ~963 proteins in 8 min neat plasma (180 samples/day) and up to 6,045 proteins with nanoparticle enrichment and 60 min runs.
• Robustness: Over 2,200 continuous injections show stable identifications (~9,625 proteins, ~96,459 peptides) with minimal drift.

Benefits and Practical Applications

The Orbitrap Astral mass spectrometer empowers large-scale clinical and cohort studies, deep single-cell analyses, robust PTM profiling, and high-throughput biomarker discovery. Its combination of speed, depth, sensitivity, and quantitation supports translational, industrial, and basic research workflows.

Future Trends and Opportunities

Ongoing developments include integration with single-cell and spatial proteomics, multi-omics platforms, advanced AI-driven data analysis, and cloud-based informatics. These extensions will further accelerate discovery and precision medicine.

Conclusion

The Orbitrap Astral mass spectrometer represents a paradigm shift in proteomic analysis, delivering unprecedented throughput, proteome coverage, sensitivity, and quantitative performance. Its robust design and comprehensive Velocity workflow enable researchers to probe biological complexity at scale and resolution previously unattainable.

References

1. Bekker-Jensen J. et al. An optimized shotgun strategy for the rapid generation of comprehensive human proteomes. Cell Syst. 2017;4(6):587–599.e4.