Agilent RapidFire 400 high-throughput mass spectrometry system

Importance of the Topic

The Agilent RapidFire 400 high-throughput mass spectrometry system addresses critical bottlenecks in drug discovery, metabolomics research, forensic testing, and clinical analysis. By eliminating conventional chromatography and using online solid-phase extraction (SPE), it accelerates sample throughput, reduces reagent consumption, and maintains analytical integrity. This capability supports faster decision-making, lower cost per sample, and more efficient use of laboratory resources.

Objectives and Study Overview

This application note introduces the design, performance, and practical advantages of the RapidFire 400 system. Key goals include:

• Demonstrating integration with triple quadrupole and time-of-flight mass spectrometers.
• Comparing throughput and data quality against conventional LC/MS workflows.
• Highlighting case studies in permeability assays, intact protein analysis, and high-capacity screening.

Methodology and Instrumentation

The RapidFire 400 employs online SPE to replace traditional HPLC separation, coupling directly to QQQ, Q-TOF, or TOF mass analyzers. Core components include:

• Temperature-controlled sample drawer (up to 64 plates; up to 138,240 samples).
• Automated plate handler and barcode reader for unattended operation.
• Three-pump fluidics based on Agilent 1260 Infinity II quaternary pumps for rapid solvent switching and method development.
• Agilent MassHunter and RapidFire Integrator/Analyzer software for data acquisition, automated quality checks, peak detection, and batch reporting.

Main Results and Discussion

• Cycle times are reduced from 38.5 h (traditional LC/MS) to 16.5 h with RapidFire-QQQ and to 4.5 h with RapidFire-TOF, achieving a 10-fold reduction in analysis time.
• Validation of PAMPA permeability assays on 81 drug candidates showed strong correlation (R2 ≈ 0.89) between RapidFire/TOF and standard LC/MS/MS data.
• Sensitivity and reproducibility were demonstrated for intact monoclonal antibodies (5–500 ng/µL) with consistent signal response in triplicate injections.
• Each SPE cartridge supports thousands of injections, reducing solvent use to <0.5 mL per sample and lowering consumable costs.

Benefits and Practical Applications

The RapidFire 400 system offers:

• High throughput: weekend-long, unattended runs of thousands of injections.
• Cost efficiency: minimal solvent consumption and reusable SPE cartridges.
• Space savings: compact footprint that can accommodate an Agilent Ultivo QQQ within the same chassis area.
• Broad applicability: drug binding assays, metabolite profiling, ADME permeability testing, protein/antibody characterization, and forensic compound screening.

Future Trends and Opportunities

Emerging directions include integration with machine-learning workflows for automated method optimization, expansion of SPE cartridge chemistries to cover new analyte classes, and further miniaturization for single-cell and microsample analysis. The RapidFire platform may also be combined with novel ionization sources and advanced high-resolution detectors to broaden its application in biopharma, environmental monitoring, and clinical diagnostics.

Conclusion

The Agilent RapidFire 400 high-throughput MS system delivers rapid, reliable, and cost-effective analysis by replacing chromatography with online SPE and high-capacity automation. Its integration with QQQ and TOF mass spectrometers, coupled with intuitive software, supports diverse applications and accelerates the path from sample to insight.

Reference

No formal literature list was provided in the original document.