Thermo Scientific Q Exactive HF-X Hybrid Quadrupole-Orbitrap MS System - PRODUCT SPECIFICATIONS

Importance of the Topic

High-resolution accurate-mass (HRAM) mass spectrometry has become a cornerstone technique in proteomics, metabolomics and biopharmaceutical analysis. Instruments capable of rapid scanning, high resolving power and precise mass measurements enable confident identification and quantitation of complex mixtures. The ability to analyze intact proteins, protein complexes, peptides and small molecules in a single run supports diverse applications in research, quality control and drug development.

Objectives and Study Overview

This application note presents the performance characteristics and technological innovations of the Thermo Scientific Q Exactive HF-X Hybrid Quadrupole-Orbitrap mass spectrometer. The aim is to highlight its speed, sensitivity, resolution and versatility, demonstrating how these features drive improvements in identification, quantitation and structural confirmation across a broad range of molecules.

Methodology and Instrumentation Used

The Q Exactive HF-X combines an ultra-high-field Orbitrap analyzer with a high-performance quadrupole for precursor selection. Key components include:

• H-ESI II ion source with dual desolvation zones and sweep gas for reduced chemical noise
• High Capacity Transfer Tube (HCTT) and Electrodynamic Ion Funnel for enhanced ion transmission
• Advanced Quadrupole Technology (AQT) featuring the HyperQuad segmented filter for sharp isolation windows
• Active Beam Guide (AABG) for improved sensitivity and robustness
• Straight multipole collision cell for accelerated Higher-Energy Collisional Dissociation (aHCD)
• Ultra-high-field Orbitrap analyzer with 240,000 resolving power (FWHM at m/z 200) and up to 40 Hz MS/MS acquisition
• Differentially pumped vacuum system (seven regions, <1 × 10⁻⁹ mbar)
• Thermo Scientific Xcalibur software for instrument control and data processing

Main Results and Discussion

The instrument achieves up to 40 Hz scan speed at 7,500 resolution and maintains sub-ppm mass accuracy (<1 ppm internal RMS). Sensitivity tests with reserpine demonstrate limits of 50 fg on column (SIM) and 100 fg in MS/MS with S/N ≥150:1. Multiplexed MS/MS enables simultaneous selection of up to ten precursors, boosting throughput in complex samples. Dynamic polarity switching, data-dependent acquisition (DDA), parallel reaction monitoring (PRM), data-independent acquisition (DIA) and all-ion fragmentation (AIF) modes provide flexible workflows for discovery, targeted quantitation and structural confirmation. A dedicated BioPharma option extends the mass range to m/z 8,000 and adds intact protein and complex analysis modes.

Benefits and Practical Applications

The Q Exactive HF-X delivers:

• Unmatched speed for deep proteome coverage and high-throughput screening
• Exceptional resolving power for confident isobar separation and structural elucidation
• Superior sensitivity for low-abundance analytes in complex matrices
• Versatile acquisition methods supporting both qualitative and quantitative experiments
• BioPharma workflows for intact protein characterization, subunit analysis and native complex studies

These capabilities facilitate research in cell biology, biomarker discovery, biotherapeutics development and small molecule quantitation.

Future Trends and Possibilities

Continued advances in Orbitrap technology will likely push resolving power, scan speed and mass range further. Integration with advanced automation, AI-driven method optimization and real-time data analysis could streamline workflows and enhance reproducibility. Expanding the capabilities for native and top-down proteomics, as well as ion mobility integration, will open new avenues for structural biology and multi-omics studies.

Conclusion

The Q Exactive HF-X Hybrid Quadrupole-Orbitrap MS system represents a state-of-the-art platform for high-resolution, high-throughput analysis of proteins, peptides and small molecules. Its combination of speed, sensitivity, resolution and versatile acquisition modes addresses the growing demands of academic and industrial laboratories, supporting deeper insights into complex biological systems and robust quantitation in regulated environments.