Shimadzu LC World Talk 2006

Importance of the Topic

High-throughput liquid chromatography (Fast LC) offers significant advantages for analytical workflows, enabling rapid separation without sacrificing resolution or requiring prohibitively high system pressures. This capability is essential in pharmaceuticals, environmental monitoring, food safety, and biotech, where sample throughput and analytical performance directly impact productivity and data quality.

Study Aims and Overview

This whitepaper examines innovative Fast LC approaches using Shimadzu’s Prominence HPLC system and novel Shim-pack XR-ODS columns packed with uniform 2.2 µm particles. It also explores elevated temperature operation and evaluates the Shimadzu LCMS-IT-TOF for high-resolution, MSn analysis of amoxicillin.

Methodology and Instrumentation

• HPLC configuration: Prominence series with LC-20AD pumps, SIL-20A autosampler, CTO-20AC/CRB-6A column ovens.
• Columns: Shim-pack XR-ODS (2.2 µm, 3 mm × 75–100 mm) and ZirChrom-PBD for elevated-temperature studies.
• Fast LC conditions: Optimized flow rates up to 3.3 mL/min, gradient cycles as short as 23 s, temperature up to 125 °C.
• Mass spectrometry: Shimadzu LCMS-IT-TOF with compressed ion injection, argon cooling, MSn capability (up to MS10), 100 ms polarity switching.

Main Results and Discussion

• 2.2 µm packing achieves similar efficiency to sub-2 µm columns but at half the backpressure, enabling longer columns and higher plate counts under 60 MPa.
• Elevated temperature LC (up to 125 °C) reduces viscosity, lowers backpressure by ~40 %, and accelerates separations via enhanced mass transfer (reduced HETP at high flow rates).
• Fast LC demonstrations: Seven alkylphenones separated at 1.5 mL/min with <0.1 % RSD in retention time; PTC-amino acids in <4 min; DNP-aldehydes/ketones in <5 min.
• Shimadzu Prominence system achieved 23-second gradient cycle without overlapping injections, with excellent peak area and retention reproducibility.
• LCMS-IT-TOF provided high mass accuracy (<5 ppm) for amoxicillin and its characteristic fragments (e.g., loss of NH3, β-lactam ring cleavage) without internal standards.

Benefits and Practical Applications

The combination of 2.2 µm columns, rapid temperature control, and robust HPLC hardware delivers:

• High sample throughput with routine cycle times under 30 s.
• Improved chromatographic resolution without specialized ultra-high-pressure systems.
• Versatile operation for both fast and conventional methods on a single platform.
• Accurate MSn and high-resolution mass data for structural confirmation in pharmaceutical and food analysis.

Future Trends and Opportunities

• Expansion of elevated-temperature LC to novel stationary phases and solvent systems.
• Integration of AI-driven software for method optimization and automated troubleshooting.
• Further miniaturization and automation of Fast LC–MS workflows for clinical and environmental applications.
• Enhanced coupling of Fast LC with orthogonal detectors for comprehensive profiling of complex samples.

Conclusion

Shimadzu’s Fast LC solution—combining Prominence HPLC, 2.2 µm Shim-pack XR-ODS columns, elevated-temperature operation, and LCMS-IT-TOF—achieves high-resolution separations at high throughput with moderate pressures. This integrated approach supports diverse analytical challenges in pharmaceutical, food safety, and biotech laboratories, delivering reliable, reproducible, and fast results.

References

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