High-Throughput Amino Acid Analysis using Tecan Automated Preparation

Importance of the Topic

Amino acids are vital biomolecules that define protein structure and function. Accurate profiling supports quality control in pharmaceutical development and optimizes nutritional conditions in cell culture media. High throughput methods address growing demands for reproducible and efficient analysis.

Objectives and Overview

This study evaluates the equivalence of manual versus automated sample preparation using a Tecan Freedom EVO 100/4 platform combined with Waters AccQ·Tag Ultra derivatization and a specialized Amino Acid Cell Culture Standard Kit. The goal is to demonstrate comparable performance in precision, accuracy, and linearity while highlighting throughput gains.

Methodology

Both workflows use pre-column derivatization with the AccQ·Tag Ultra kit followed by UPLC separation and UV detection at 260 nm. Robustness was assessed across three lots of reagents and columns and by four different analysts. Sample preparation covered three concentration panels (10, 200, 400 µM) with triplicate injections to determine retention time, peak area, recovery, and calibration linearity over 0.5–500 µM.

Instrumentation

• ACQUITY UPLC H-Class Bio system with tunable UV detector
• AccQ·Tag Ultra 1.7 µm 2.1×100 mm column
• AccQ·Tag Ultra Derivatization Automation Kit and Amino Acid Cell Culture Standard Kit
• Tecan Freedom EVO 100/4 automated liquid handler
• Empower 3 chromatography data system

Main Results and Discussion

Automated preparation matched manual methods in retention time precision (CV ≤0.3%) and improved peak area repeatability (CV ≤1.8% vs ≤2.8%). Recovery for automation met ±20% at low concentration and ±15% at mid/high levels. Linearity was excellent with r2>0.995 across all amino acids. Representative chromatograms confirmed resolution of 17 hydrolysate and nine cell culture amino acids.

Benefits and Practical Applications

• Reduction of hands-on time: preparation of 96 samples in under one hour
• Minimized human error and cross-contamination
• Elimination of analyst-to-analyst variability and streamlined method transfer
• Freeing analysts for other laboratory tasks

Future Trends and Potential Applications

Integration of automated amino acid analysis with laboratory information management systems and AI-driven data review will further enhance throughput and quality assurance. Expansion to multiplexed assays and miniaturized formats may enable broader adoption in QC and research settings. Ongoing development of new derivatization reagents could extend the method to modified amino acids and related metabolites.

Conclusion

Automation of AccQ·Tag derivatization on the Tecan Freedom EVO 100/4 achieves equivalent precision, accuracy, and linearity to manual workflows while significantly increasing efficiency and reproducibility. This platform is recommended for laboratories requiring high-throughput standardized amino acid analysis.

References

• Salazar A Keusgen M von Hagen J Amino Acids in the Cultivation of Mammalian Cells Amino Acids 2016 48 5 1161–1171
• Saitoh S Yoshimori T Fully Automated Laboratory Robotic System for Automating Sample Preparation and Analysis to Reduce Cost and Time in Drug Development Process JALA 2008 13 5 265–274