Separate, purify, go! Vanquish Analytical Purification LC System

Significance of the Topic

Purification by fractionation is essential in drug discovery, manufacturing, and quality control for isolating pure compounds from complex matrices. Efficient chromatographic separation and precise fraction collection underpin the identification of active substances, impurity profiling, and preparation of reference standards, directly impacting the reliability of downstream analyses.

Objectives and Overview of the Study

This article evaluates the Thermo Scientific™ Vanquish™ Analytical Purification LC System, illustrating how its integrated design combines HPLC/UHPLC separation power with highly accurate fractionation. The goals are to demonstrate system performance, fluidic innovations, and application flexibility for diverse laboratory workflows.

Methodology and Instrumentation

The system integrates a Vanquish LC platform (Core HPLC, Flex UHPLC or Horizon UHPLC) with an automated Vanquish Fraction Collector featuring:

• Low–volume, fast‐switching fraction collection valve to maintain peak resolution.
• Viper™ Fingertight Fitting fluidics to minimize dead volumes.
• Trigger options based on time or detector signal (UV, fluorescence, charged aerosol, MS via flow split).
• Adjustable needle positioning and configurable movement paths (horizontal, vertical, sawtooth) for various vessel formats.
• Automated delay volume determination using an air‐bubble method within Chromeleon CDS for precise valve timing.
• Chromeleon software with a stepwise method wizard and fractionation simulation for rapid method development.

• Vanquish Core HPLC: up to 10 mL/min.
• Vanquish Flex UHPLC: up to 8 mL/min, biocompatible.
• Vanquish Horizon UHPLC: from 50 µL/min for low‐abundance analytes.

Main Results and Discussion

Experimental evaluation showed:

• Preservation of chromatographic resolution during fractionation due to minimized internal delay volumes.
• High recovery (>95%) and purity across flow rates and trigger modes.
• Minimal carry‐over through optimized flush, rinse, and wash cycles.
• Delay volume determination within ±5% variance, improving fraction valve switching accuracy.
• Seamless handling of multiple container formats (vials, well plates) in a temperature‐ and light‐controlled fraction compartment.
• Intuitive Chromeleon interface enabling real‐time fractionation preview and reduced method setup time.

The sawtooth needle path minimized cross‐contamination and dripping during high‐speed movements.

Benefits and Practical Applications of the Method

• Rapid isolation of target compounds for structural elucidation, bioassays, and scale‐up studies.
• Enhanced impurity profiling during process development and QC workflows.
• Streamlined purification workflows by unifying separation, fractionation, and data processing.
• Customizable fraction sizes tailored to analytical or preparative needs.
• Effortless method transfer between analytical and preparative modes and across varying sample volumes.

Future Trends and Potential Applications

Emerging directions include:

• Further miniaturization for single‐cell and trace‐level fractionation.
• AI‐driven peak detection and adaptive fractionation in real time.
• Integration with orthogonal purification techniques (SPE, SFC) in unified workflows.
• Compatibility with ultra‐high‐throughput formats (e.g., 1536‐well plates).
• Expanded use in targeted metabolomics and proteomics for selective compound isolation.

Conclusion

The Vanquish Analytical Purification LC System delivers pinpoint fractionation across a broad range of chromatographic conditions. Its engineered fluidics, versatile fraction collector, and intuitive software collectively ensure high recovery, minimal carry‐over, and preserved resolution, accelerating purification workflows in research and production settings.

References

1. Thermo Scientific Technical Note 72940: Principles of fraction collection using the Vanquish HPLC and UHPLC systems.