Using the Fraction Collector in Empower 3 Environment - Technical Note

Importance of the Topic

Automated fraction collection is essential for preparative chromatography, method development, and sample purification workflows. Integrating fraction collectors with chromatographic systems streamlines downstream processing, improves reproducibility, and supports complex applications such as bioanalysis, reaction monitoring, and compound isolation.

Objectives and Overview of the Guide

This technical guide aims to detail the configuration and operation of Agilent fraction collectors within Waters Empower 3 software. It covers hardware installation, software preconfiguration, method setup, routine sample runs, configuration changes, and known limitations to ensure a robust and vendor-agnostic fraction collection workflow.

Methodology and Instrumentation Used

The workflow employs Agilent LC modules (fraction collectors G1364A–D and G5664A) connected via CAN bus to a Waters Empower Data System. Firmware requirements must meet or exceed vendor specifications. The PreConfiguration Utility, provided with Agilent ICF Support, is used locally or via the Empower Configuration Manager to detect and configure modules. Key instrumentation elements include:

• Agilent 1100/1260 Infinity fraction collectors (analytical, preparative, micro-scale, bio-inert)
• Agilent LC modules with minimum firmware A.06.53 for fraction collectors
• Waters Empower 3 software and Agilent ICF Support packages v2.1 HF1 through v3.2
• CAN communication cables and optional G1390A Universal Interface Box for MS coupling

Main Procedures and Discussion

Installation and configuration consist of:

• Hardware setup: power down LC, connect fraction collector via CAN cable, load trays or plates, power up system
• Module discovery: launch PreConfiguration Utility locally or through Empower Configuration Manager, auto-detect LC modules and fraction collector
• Software configuration: define communication settings, peak detectors, collection options, inner capillary dimensions, needle type, plate definitions, and delay volume calibration
• Method creation: in Empower method editor, select fraction trigger mode (peak- or time-based), define fraction count or time slices, set start location syntax (e.g. P1-A-1), and configure advanced parameters such as rinse, auxiliary limits, and peak thresholds
• Sample run: verify tray presence and closure, assign plates or tube parameters via module options or tile context menu, and execute sequence
• Configuration changes: replacing trays or plate types triggers reconfiguration; a system reboot and new auto-configuration sync hardware and software

Benefits and Practical Applications

Combining Agilent fraction collectors with Empower software offers:

• Flexible fractionation strategies (discrete or continuous flow, peak- or time-based)
• Support for diverse consumables (vials, tubes, wellplates) and custom plate formats
• Automated synchronization between hardware and software to minimize user error
• Enhanced workflow reproducibility and traceability in QA/QC, pharmaceutical development, and preparative separations

Future Trends and Potential Uses

Emerging directions include mass-based fraction collection integration, real-time decision making via advanced detectors, AI-driven method optimization, high-throughput microfluidic fractionation, and seamless coupling with downstream analytical platforms for automated sample processing.

Conclusion

This guide provides a clear roadmap for setting up and using Agilent fraction collectors within Empower 3, addressing hardware installation, software configuration, method development, routine operation, and troubleshooting. Adhering to these best practices ensures reliable fractionation and maximizes system performance.

Reference

• Agilent Technologies D0002925 Edition 04/2020, Technical Guide for configuration and use of Agilent Fraction Collectors with Waters Empower
• Waters Document TECN134936402, Using the Agilent PreConfiguration Utility with Agilent ICF Support Version 2.2