Agilent Infinity UHPLC Nanodapter

Significance of the topic

Nano-flow liquid chromatography (nano-LC) is critical for high-sensitivity proteomic analyses, enabling detection of low-abundance peptides from limited sample amounts. The agility to switch between analytical and nano-flow regimes enhances lab productivity and broadens method applicability, particularly in biomarker discovery and quality control scenarios.

Objectives and Study Overview

This data sheet introduces the Agilent Infinity UHPLC Nanodapter, designed to adapt the 1290 Infinity II Binary Pump for nano-LC applications. It outlines system reconfiguration procedures, performance parameters, and compatibility with the G1992A Nanospray Ion Source for proteomics workflows.

Methodology and Instrumentation

The Nanodapter passively splits the pump flow into 100–1,000 nL/min for columns 0.050–0.150 mm I.D. Key features include:

• In-line electronic flow sensor with PC software for real-time monitoring
• Optional divert valve for rapid transfer to a guard column
• Maximum operating pressure up to 1,000 bar (14,500 psi)
• Switch time between conventional and nano-flow modes under 30 minutes

Key Results and Discussion

Retention time precision is maintained below 0.25 min (SD), and system dead volume yields approximately 5 min delay at 350 nL/min. Base peak chromatograms of 500 ng E. coli digest demonstrate robust performance at 340 nL/min. Extending gradients to 120 min with 500 ng injections produced ~1,260 proteins and ~8,700 unique peptides per run, indicating high proteome coverage and reproducibility.

Benefits and Practical Applications

• Enhanced sensitivity for low-level analytes in proteomics
• Flexible workflow transitions between analytical and nano scales
• Minimal carryover and reproducible retention times
• High-pressure capability supports efficient sub-2 µm column chemistries

Future Trends and Opportunities

Integration with automation platforms and microfluidic sample preparation will drive higher throughput. Developments in column technology and ultra-low flow ESI sources may further improve detection limits, enabling single-cell proteomics and advanced metabolomics studies.

Conclusion

The Agilent Infinity UHPLC Nanodapter offers a versatile and rapid solution for converting high-flow UHPLC systems to nano-LC, delivering precise flow control, high pressure tolerance, and reproducible chromatographic performance vital for modern proteomic and bioanalytical applications.

Used Instrumentation

• Agilent 1290 Infinity II Binary Pump
• Agilent Infinity UHPLC Nanodapter module
• G1992A Nanospray Ion Source
• Agilent AdvanceBio Peptide Map C18, 0.075 × 150 mm, 2.7 µm column