Performance Characteristics of the Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector

Importance of the Topic

Preparative-scale HPLC fraction collection is essential for isolating pure compounds in pharmaceutical, chemical and biochemical research. Achieving high recovery and purity requires precise synchronization between detector signals and the fraction collector to minimize sample loss and cross-contamination.

Objectives and Study Overview

This study evaluates the performance characteristics of the Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector under peak-based fraction collection triggered by ultraviolet (UV) detection alone, combined UV and mass-selective detection (MSD), and time-slice collection mode. Purity and recovery metrics are determined for model compounds, and the impact of fraction trigger strategies is compared.

Methodology

Samples of eight analytes were separated on a preparative C18 column (21.2×100 mm, 5 µm) with a gradient of 10–98 % acetonitrile containing 0.1 % formic acid at 25 mL/min. Peak-based collection parameters included UV threshold (50 mAU), upslope (50 mAU/s), and downslope (5 mAU/s). Mass-based triggers used target ion masses with 50 000 cps thresholds. Time-slice mode employed 0.06 min slices triggered by lower thresholds to segment peaks for selective pooling. Re-analysis of fractions was performed on an analytical C18 column (4.6×50 mm, 5 µm) to quantify purity (UV 254 nm) and recovery.

Instrumentation

• Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector
• Agilent 1260 Infinity II Binary LC with diode array detector
• Agilent 1260 Infinity II Quaternary Pump for MSD makeup
• Agilent 1260 Infinity Dual-Loop Autosampler
• Agilent 6150 Single Quadrupole LC/MS (Jet Stream)
• Controlled by Agilent OpenLAB CDS ChemStation

Results and Discussion

Peak-based UV-triggered collection yielded purities of 98.9–99.6 % and recoveries of 97.5–100 % for three closely eluting compounds. Combining UV and MSD triggers improved purity slightly (99.2–100 %) with recoveries above 96 %, at the expense of a marginally narrower collection window. Time-slice collection segmented peaks into multiple fractions, achieving pooled purities of ~99 % with recoveries above 92 % despite unavoidable dead time during tube switching.

Practical Benefits and Applications

The Agilent 1290 Infinity II preparative collector offers flexible fractionation modes to balance purity and yield. UV-only, UV+MSD, and time-slice strategies enable users to optimize workflows for complex separations, targeted compound isolation, and impurity profiling in preparative HPLC settings.

Future Trends and Applications

Advances may include real-time data processing for adaptive trigger settings, integration of additional detection modes, and miniaturized fraction collectors for microflow systems. Machine learning could further refine fraction coordination and predictive yield optimization.

Conclusion

The Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector delivers high-purity, high-recovery fractionation in peak-based and time-slice modes. Its precise delay calibration and flexible triggering ensure reliable performance for preparative HPLC purification.

References

1. Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector – Collect Each and Every Drop, Agilent Technologies Brochure 5991-7225EN, 2016.
2. Rieck F. Time-, Peak-, and Mass-Based Fraction Collection with the 1290 Infinity II Preparative Open-Bed Fraction Collector. Agilent Technical Overview, 5991-7654EN, 2016.