THE HIGH THROUGHPUT PREPARATION OF AMINO ACIDS USING WATERS TECHNOLOGIES NEW KAIROS AMINO ACID 500+ AUTOMATION KIT ON THE FREEDOM EVO 100 PLATFORM

Importance of Topic

Amino acid profiling plays a critical role in clinical diagnostics, pharmaceutical development, food quality control and fundamental research. Traditional manual workflows for sample derivatization and preparation are time consuming, labor intensive and prone to variability. By adopting a high-throughput automated solution, laboratories can significantly reduce hands-on time, improve reproducibility and accelerate data turnaround.

• Amino acids serve as biomarkers and quality indicators in many industries.
• Manual derivatization workflows are bottlenecks for large-scale studies.
• Automation enhances consistency, throughput and traceability.

Aims and Study Overview

This work describes the development and evaluation of Waters Technologies’ Kairos Amino Acid 500+ automation kit paired with a custom script on the Tecan Freedom EVO 100 platform. Key objectives included:

• Establish a full-service kit supporting calibrators, QC, internal standard and reagents.
• Implement barcode tracking and file conversion for seamless integration with MassLynx software.
• Demonstrate preparation of 96 samples in under two hours with average CV <2.6%.
• Compare automated performance against the established manual protocol over three days.

Methodology and Instrumentation

The automated workflow integrates sample handling, derivatization and dilution steps in a single script requiring minimal user intervention (centrifuge pause and basic inputs). Key components:

• Automation platform: Tecan Freedom EVO 100 with barcode reader and custom liquid classes.
• Kit: Waters Kairos Amino Acid 500+ containing all needed reagents, calibrators, QCs, internal standard and consumables.
• Analytical system: Waters ACQUITY UPLC I-Class coupled to XEVO TQ-S micro tandem MS.
• Throughput: 96 sample preparations completed in under two hours.

Main Results and Discussion

The automated method delivered accuracy and precision comparable to manual preparation across 20 essential and non-essential amino acids over three days:

• Average accuracy ≥87.5% for all kit amino acids, with individual values often exceeding 95%.
• Precision (mean CV) ≤4.5% across five concentration levels (20–2500 µM).
• Calibration curves showed linearity (r2≥0.99) with acceptance criteria met at all points.
• Automated and manual protocols produced equivalent performance profiles, confirming robustness.

Benefits and Practical Applications

Implementing the automated kit and script offers several advantages:

• Substantial reduction in operator time and technical skill requirements.
• Minimized risk of pipetting errors and sample mix-ups via barcode tracking.
• High reproducibility supports regulatory and quality assurance demands.
• Scalable to routine workflows in clinical, pharmaceutical and food testing laboratories.

Future Trends and Applications

Ongoing developments and potential expansions include:

• Porting scripts to additional robotic platforms to broaden accessibility.
• Extension of amino acid panels or integration with other bioanalytical assays.
• Enhanced software connectivity for real-time data management and LIMS integration.
• Potential adoption in diagnostic laboratories as workflows reach clinical validation.

Conclusion

The Waters Kairos Amino Acid 500+ automation kit on the Freedom EVO 100 platform provides a turnkey solution for high-throughput amino acid sample preparation. It matches manual methods in accuracy and precision while dramatically improving efficiency and reproducibility. Future work will focus on cross-platform compatibility and expanded assay capabilities.

References

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