The Complete and Easy Guide to Configuring Your Thermo Scientific™ NanoLC for Mass Spectrometric Analysis

Significance of the topic

The coupling of nano-scale liquid chromatography (nano-LC) with mass spectrometry (MS) is a cornerstone technique in proteomics, metabolomics, and trace analysis. Optimizing the configuration of nano-LC systems enhances sensitivity, reproducibility, and throughput in complex sample analyses.

Objectives and Overview of the Guide

This application note aims to provide a clear, step-by-step framework for selecting, configuring, and maintaining Thermo Scientific™ nano-LC platforms (EASY-nLC 1200/1000, EASY-nLC II, UltiMate™ 3000 RSLCnano) for MS detection. It presents decision trees for system setup, detailed schematics of fluidic layouts, consumable part numbers, and best practices for fluidic connections and system pressure management.

Methodology and Instrumentation

The guide is structured around three key nano-LC systems:

• EASY-nLC 1200/1000: supports up to 1200 bar, multiple pre-column and analytical column configurations, liquid junction options, and silica or packed emitters.
• EASY-nLC II: a 1000-bar system with selection trees for pre-column, analytical column, and emitter assemblies.
• UltiMate™ 3000 RSLCnano: high-throughput RSLCnano platform with EASY-Spray connection kits, heated columns, and direct-injection options.

Each section includes visual selection trees, detailed tubing and union schematics, tubing sleeves, liquid junction kits, and emitter types with catalog numbers.

Main Results and Discussion

The note delivers comprehensive configuration maps, enabling users to match column inside diameters, emitter styles, and pressure limits to specific analytical goals. It highlights the importance of minimizing dead volume, correct installation of nanoViper/Viper fittings, and applying pressure limits in instrument control software. Maintenance guidelines specify replacement intervals for unions, sleeves, and capillaries to ensure consistent performance.

Benefits and Practical Applications

The structured approach reduces trial-and-error, accelerates system setup, and improves data quality by maintaining optimal fluidic continuity. Laboratories benefit from reduced downtime, standardized procedures, and clear part references for routine maintenance and troubleshooting.

Future Trends and Potential Applications

Emerging trends include integration of higher-pressure nano-LC systems with microfluidic interfaces, automated column switching, and on-the-fly pressure optimization. Advances in emitter materials and biocompatible flow paths will further expand applications in cell signaling and single-cell proteomics.

Conclusion

This guide consolidates best practices for configuring Thermo Scientific nano-LC platforms for mass spectrometric analysis. By following the provided selection trees, schematics, and maintenance protocols, users can achieve reliable, high-sensitivity separation with minimal dead volume and robust system performance.

Used Instrumentation

• Thermo Scientific™ EASY-nLC 1200 and 1000
• Thermo Scientific™ EASY-nLC II
• Thermo Scientific™ UltiMate™ 3000 RSLCnano
• Thermo Scientific™ EASY-Spray™ and Nanospray™ Flex sources
• NanoViper and Viper fittings, Liquid Junction kits, MicroTight® unions

References

Thermo Scientific™. The Complete and Easy Guide to Configuring Your Thermo Scientific™ Nano LC for Mass Spectrometric Analysis. December 2015.