Thermo Scientific UltiMate 3000 RSLCnano UHPLC System - PRODUCT SPECIFICATIONS

Significance of the Topic

The development of robust low-flow UHPLC systems has transformed proteomics, metabolomics and biopharmaceutical analyses by delivering enhanced sensitivity, peak capacity and reproducibility in LC-MS workflows. A versatile nanoflow platform enables researchers to achieve reliable detection of trace analytes while maintaining high throughput and method flexibility.

Objectives and Study Overview

This application note presents the Thermo Scientific UltiMate 3000 RSLCnano system, designed to address the demands of routine and advanced low-flow UHPLC-MS applications. Key goals include demonstrating retention time precision, modular instrument configuration, pressure performance and compatibility with state-of-the-art mass spectrometers.

Methodology and Instrumentation

The modular system integrates several core components to support nano, capillary and micro flow rates:

• Binary high-pressure gradient pumps with dedicated nano, capillary or micro ProFlow flow meters
• Optional ternary micro pump for sample preconcentration and multidimensional gradients
• Column compartment with precise temperature control and low dead-volume switching valves
• Thermostatted autosampler offering full-loop, partial-loop and low-dispersion injection modes
• Variable wavelength UV/Vis detector optimized for low-volume flow cells
• Solvent rack with integrated degasser for stable mobile phase delivery

Main Results and Discussion

Performance benchmarks highlight the system’s capabilities:

• Flow range from 50 nL/min to 50 µL/min (and up to 2.5 mL/min with micro pump) with pressures up to 900 bar
• Retention time repeatability ≤0.2% RSD for 30-minute gradients
• System delay volumes below 350 nL in preconcentration mode and gradient delay <25 nL
• Column temperature stability within ±0.1 °C and rapid heating from 35 to 65 °C in 12 minutes
• Ultra-low dead volume switching valves enabling two-column workflows and uninterrupted nanospray operation
• Autosampler injection precision <0.4% RSD for 1 µL full-loop injections and carryover below 0.02%

Benefits and Practical Applications

The UltiMate 3000 RSLCnano system offers:

• High sensitivity and peak capacity for nano LC-MS analyses using 75 cm×75 µm columns
• Enhanced reproducibility and retention time accuracy via ProFlow technology
• Flexible configuration to tailor flow rates, gradient formation and multi-dimensional separations
• Fast startup, automated leak monitoring and GLP features for routine QA/QC environments

Future Trends and Opportunities

Emerging directions in low-flow UHPLC include integrating artificial intelligence for method optimization, coupling parallelized nanoflow channels for higher throughput, advancing novel monolithic and microfabricated columns, and deeper automation of sample preparation and multi-omic workflows. Continued hardware and software innovation will further enhance data quality and laboratory efficiency.

Conclusion

The Thermo Scientific UltiMate 3000 RSLCnano system represents a comprehensive low-flow UHPLC solution, combining precision fluidics, modular expandability and seamless MS compatibility. Its performance attributes position it as a key tool for researchers seeking high-resolution, reproducible separations in proteomics, metabolomics and biopharmaceutical analysis.