High Efficiency in Workflow from Preparative HPLC to Analytical HPLC by Nexera™ Prep System

Importance of the Topic

The integration of preparative and analytical liquid chromatography–mass spectrometry (LC-MS) workflows addresses a critical need in pharmaceutical and industrial analytics for rapid compound purification and purity confirmation. Automated MS-triggered fractionation and seamless reinjection for analysis significantly reduce manual intervention, turnaround time, and risk of contamination.

Objectives and Study Overview

This study presents a convertible LC-MS system (Nexera Prep) combining preparative separation and analytical confirmation within a single platform. The goal is to demonstrate high-efficiency workflow from preparative fractionation to purity assessment using MS triggers and high-throughput analytical reanalysis.

Methodology and Instrumentation

• System Configuration: LH-40 liquid handler with parallel preparative and analytical flow paths controlled by LabSolutions™ workstation.
• Preparative LC: Shim-pack Scepter C18 column (75 × 30 mm, 5 μm), dual LC-20AP pumps at 40 mL/min, gradient from 10 % to 90 % acetonitrile (1 % formic acid), UV detection at 250 nm, and LCMS-2020 for MS detection.
• Splitter Piping: Seven-segment PEEK/PTFE tubing split fraction stream between MS detector and fraction collector.
• Analytical LC: Shim-pack Scepter C18 column (50 × 4.6 mm, 3 μm), LC-20AB analytical pump at 2.5 mL/min, rapid gradient (10 % to 90 % B in 2–3 min), UV detection and MS scan (m/z 50–1000).

Main Results and Discussion

• Model compounds ketoprofen and indomethacin were separated by preparative LC-MS and collected based on extracted ion chromatograms (XICs), yielding two distinct fractions.
• Automated reinjection of fractions into the analytical flow path enabled purity assessment within 3 min per sample, achieving ≥99 % purity by UV area normalization for both compounds.
• The workflow eliminated manual transfer steps, demonstrating seamless transition between preparative and analytical modes.

Benefits and Practical Applications

• Fully automated end-to-end process reduces operator workload and potential errors.
• High-throughput capability facilitates rapid screening and quality control of multiple fractions.
• Compact setup conserves laboratory space and simplifies method management.

Future Trends and Potential Applications

• Application to diverse sample types, including complex natural products and biologics.
• Integration with high-resolution mass spectrometry for enhanced selectivity and identification.
• Implementation of AI-driven method development and real-time decision algorithms for fraction collection.
• Development of micro-preparative and multiplexed systems for ultra-high throughput workflows.

Conclusion

The Nexera Prep convertible LC-MS system successfully streamlines preparative purification and analytical confirmation into a unified, automated workflow. MS-triggered fractionation combined with reinjection for high-speed analysis offers significant gains in efficiency, accuracy, and throughput, making it a valuable tool for pharmaceutical research, QA/QC operations, and industrial analytical laboratories.