Agilent 6560 Ion Mobility Q-TOF LC/MS System

Significance of the Topic

The Agilent 6560 Ion Mobility Q-TOF LC/MS system introduces a third dimension of separation by combining liquid chromatography mass spectrometry with gas-phase ion mobility
This enhancement delivers greater sensitivity selectivity and structural insight across proteomics metabolomics lipidomics glycomics and biotherapeutic development

Study Objectives and Overview

Assess the performance of the Agilent 6560 system in resolving complex mixtures and characterizing molecular conformations
Demonstrate the system’s ability to increase metabolite coverage detect low-level analytes and map pan-omic profiles
Highlight the unique ion funnel and uniform-field drift tube design underlying performance gains

Methodology and Instrumentation

The setup integrates a 1200 Infinity Series HPLC front end with a multi-segment iFunnel assembly and a uniform-field drift tube preceding a high-resolution Q-TOF analyzer
Each segment (front funnel trapping funnel drift tube rear funnel) is optimized for maximal ion transmission resulting in only two-fold signal loss versus standalone Q-TOF
Collision cross-section values are calculated directly under controlled temperature pressure and field conditions achieving <2% accuracy without external calibration
MassHunter software enhancements enable linked 3-D visualization drift-time filtering and automated CCS determination

Main Results and Discussion

Baseline separation of isobaric trisaccharides and permethylated oligosaccharides by CCS and m/z demonstrated resolving power up to 60
Drift-time resolved All Ions MS/MS in serum allowed identification of low-level peptides unreachable by conventional LC/MS
Protein conformers including IgG2 isomers were differentiated by distinct drift times revealing fine structural details at pg/mL sensitivities
Pan-omic mapping in CCS versus m/z space separated lipids peptides carbohydrates and other biomolecules in a single infusion

Benefits and Practical Applications

Enhanced peak capacity and orthogonal selectivity differentiate structural isomers conformations and charge states not resolvable by LC or MS alone
Rapid untargeted surveys support systems biology biomarker discovery food safety QA/QC and biomanufacticuring process monitoring
Reliable CCS measurements facilitate biotherapeutic characterization including glyco-isoform analysis and antibody drug conjugate profiling

Future Trends and Possibilities

Expansion of compound databases and libraries integrated with All Ions MS/MS for high-throughput structural annotation
Next-generation ion funnel and drift tube refinements to push sensitivity and throughput limits
Adoption in polymeromics environmental monitoring and real-time bioprocess control
Application of machine learning to interpret high-dimensional mobility-mass data enabling predictive molecular modeling

Conclusion

The Agilent 6560 Ion Mobility Q-TOF LC/MS system redefines analytical boundaries by delivering unmatched separation sensitivity and structural insight
Its innovative ion funnel design uniform-field drift tube and direct CCS measurement empower breakthrough discoveries across multiple omics disciplines
Coupled with advanced software pan-omic workflows researchers can now access previously hidden molecular details with confidence and ease

References

• May J.C. et al. Anal Chem 2014 86(4) 2107-2116
• Campuzano I. et al. Anal Chem 2012 84(2) 1026-1033