Time-, Peak-, and Mass-Based Fraction Collection with the Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector

Significance of the Topic

Fraction collection transforms analytical HPLC into a preparative tool, enabling isolation of target compounds for downstream processing. High flow rates (5–100 mL/min), variable fraction volumes (mL to hundreds of mL), and diverse sample loads demand flexible collector configurations. Precise timing, high throughput, and multi-signal triggers are key for efficient purification in pharmaceutical, environmental, and industrial applications.

Objectives and Study Overview

This Technical Overview examines the enhanced features of the Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector. It highlights new fraction modes—time-, peak-, and mass-based—combined with volume or time slices, and advanced UV/MS triggering logic (AND, OR, AND/OR). Representative applications demonstrate performance across scouting gradients and optimized separations.

Methodology and Used Instrumentation

The preparative setup included:

• Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector
• Agilent 1260 Infinity II Preparative Pump Cluster (G1361A/G1391A)
• Agilent 1260 Infinity Quaternary Pump for MSD make-up
• Agilent 1260 Dual-Loop Autosampler with 50 μL and 5 mL loops
• Agilent 1260 Diode Array Detector (3 mm prep cell)
• Agilent 6150 Single Quadrupole LC/MS with Jet Stream
• Active Flow Splitter and OpenLAB CDS ChemStation control software
• ZORBAX SB-C18 preparative (21.2 × 100 mm) and analytical (4.6 × 50 mm) columns

Samples comprised eight model compounds (acetaminophen, sulfamerazine, caffeine, methyl/ethyl/propyl parabens, sulfadimethoxine, benzyl-4-hydroxybenzoate) in DMSO, analyzed with scouting and optimized gradients.

Main Results and Discussion

Time‐based volume slices collected nine crude fractions (0–7 min, 21 mL each) for initial scouting. Peak-based with volume recovery isolated eight distinct compounds while preventing sample loss. Peak-based time slices (0.04 min) “clipped” overlapping peaks into discrete fractions for purity assessment. Targeted peak collection (4.00–7.50 min) using slope and threshold parameters separated close eluting peaks. UV/MS triggers with logical AND provided highest specificity for a target mass (310 Da), while OR and AND/OR modes balanced sensitivity and peak width, ensuring comprehensive recovery without premature cut-off.

Benefits and Practical Applications

Enhancements address preparative challenges:

• Modular fraction bed with six containers for tubes of various diameters and lengths
• Three accessible drawers allowing in-run exchange without loss
• Built-in detector-to-collector delay calibration
• Eight combined fraction modes adapting to sample scarcity or purity demands
• Multi-signal logic for complex matrices and coelutions

These features streamline workflows in purification, sample re-analysis, pooling, and solvent evaporation.

Future Trends and Opportunities for Use

Emerging directions include integration with high-resolution MS, AI-driven method optimization, real-time analytics, and automated fraction quality assessment. Applications will expand in biopharmaceutical purification, natural product isolation, metabolomics, and regulated QA/QC processes.

Conclusion

The Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector combines high capacity, flexible tube handling, precise calibration, and advanced fraction modes—time-, peak-, and mass-based with Boolean trigger logic—to meet diverse preparative demands. Its versatile configuration and intelligent software support efficient, specific, and reproducible isolations.

References

1. Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector – Collect Each and Every Drop, Agilent Technologies Brochure, publication number 5991-7225EN, 2016.
2. Rieck F. Performance Characteristics of the 1290 Infinity II Preparative Open-Bed Fraction Collector – Purity and Recovery of Collected Compounds, Agilent Technologies Technical Overview, publication number 5991-7655EN, 2016.