Optimized reproducibility using a state of the art UHPLC system for the analysis of catechins found in tea

Importance of the topic

Catechins are key antioxidant compounds abundant in tea and other food and beverages. Accurate and reproducible chromatographic analysis of these labile molecules is critical for quality control, regulatory compliance, and research into health benefits. Ensuring seamless transfer of UHPLC methods between systems reduces revalidation effort and enhances laboratory efficiency.

Objectives and study overview

This study aimed to demonstrate the transfer of a catechin separation method from a Thermo Scientific UltiMate 3000 RS UHPLC system to a Thermo Scientific Vanquish Flex UHPLC system without altering chromatographic parameters. It also evaluated the suitability of the Thermo Scientific Acclaim RSLC 120 C18 column for seven common tea catechins and compared system reproducibility in retention time, peak width, and peak area.

Methodology and instrumentation

Sample preparation involved dissolving seven catechin standards (15 µg/mL each) in 0.1 % TFA aqueous mobile phase. A gradient from 100 % aqueous to 100 % acetonitrile over 25 minutes at 0.45 mL/min was employed at 25 °C using a 150 × 2.1 mm, 2.2 µm Acclaim RSLC 120 C18 column. UV detection was performed at 280 nm.

• UltiMate 3000 RS UHPLC system: LPG-3400RS pump, WPS-3000RS autosampler, TCC-3000RS column oven, DAD-3000RS detector with 13 µL flow cell.
• Vanquish Flex UHPLC system: Quaternary Pump F, Split Sampler FT, Column Compartment H with active pre-heater, Diode Array Detector HL with 10 mm LightPipe flow cell.
• Data acquisition: Chromeleon CDS 7.2 SR4; Virtuoso Vial Identification System for automated sample tracking.

Main results and discussion

Both systems achieved baseline separation of all seven catechins within 13 minutes under identical gradient conditions and column configuration. Retention times were closely matched, requiring no method adjustment for dwell volume differences. Evaluating 15 sequential injections, the Vanquish Flex system improved reproducibility by at least 50 % relative to the UltiMate 3000 system in terms of retention time RSD, peak width at half height RSD, and peak area RSD. Enhanced precision is attributed to SmartInject autosampler technology and optimized low‐pressure mixing pump design.

Benefits and practical applications

• Reliable method transfer reduces validation time and cost across laboratories.
• High reproducibility ensures robust QA/QC workflows for food and beverage analytics.
• The Acclaim RSLC 120 C18 column delivers high efficiency and resolution for catechin profiling.
• Improved autosampler and pump performance extends column life and data confidence.

Future trends and potential applications

Advances may include tighter integration of UHPLC systems with laboratory information management systems, expanded use of quaternary gradients for complex sample matrices, and coupling with mass spectrometry for structural confirmation of catechin derivatives. Emerging detectors and microfluidic technologies will further reduce analysis times and solvent consumption, enhancing sustainability.

Conclusion

The successful method transfer from an UltiMate 3000 RS to a Vanquish Flex UHPLC system using the Acclaim RSLC 120 C18 column demonstrates superior reproducibility and reliability without altering chromatographic conditions. This streamlined approach supports efficient catechin analysis in research and quality control environments.

References

• No specific literature references were provided in the original text.