Automated HPLC method development and robustness tests for abacavir, lamivudine, dolutegravir and their related compounds in Triumeq drug produ

Significance of Topic

Developing reliable HPLC methods for combination therapies such as Triumeq is critical to ensure product quality and regulatory compliance. Automated method development accelerates this process, reduces human error, and supports Analytical Quality by Design (AQbD) principles by systematically exploring chromatographic parameters.

Study Objectives and Overview

This study aimed to establish a fully automated workflow for HPLC method development and robustness testing of abacavir, lamivudine, dolutegravir, and their related impurities in the Triumeq drug product. The key goals were:

• Automate method scouting, optimization, and robustness assessment.
• Demonstrate significant time savings compared with manual development.
• Achieve chromatographic resolution ≥2.0 and acceptable peak symmetry.

Methodology and Instrumentation

Automated chromatography was performed using ChromSwordAuto Chromeleon Connect software integrated with a Thermo Scientific Vanquish Core HPLC system and automated column switching valves. The workflow comprised:

• Method scouting on five 4.6×150 mm, 3 µm columns (C18, phenyl, PFP chemistries).
• Eluent systems ranging from 0.1% formic acid to ammonium formate buffers (pH 2.7–7.1) and methanol/acetonitrile gradients.
• Rapid and fine optimization experiments varying flow rate, column temperature (25–40 °C), injection volume, and pH.
• Robustness testing via a full factorial design exploring ±5% organic concentration, ±5 °C temperature, ±0.5 pH units, and gradient breakpoint time variation.

Sample preparation involved grinding Triumeq tablets, extracting with 50% methanol, spiking with impurities at defined levels, sonication, and centrifugation. Data acquisition was managed by Chromeleon CDS version 7.3.

Main Results and Discussion

The automated workflow completed scouting, optimization, and robustness testing in approximately 136.9 hours of instrument time with minimal analyst intervention, representing about one-fifth of the time required by traditional manual methods. Key outcomes included:

• Selection of the Acclaim 120 C18 column with pH 5.0 aqueous phase and methanol for optimal separation.
• Final gradient method delivering baseline resolution (USP ≥ 2.0) for all 20 analytes and peak asymmetry factors between 0.9 and 2.1.
• Identification of a robust design space for column temperature, mobile phase pH, and gradient breakpoint that ensures consistent performance.

Benefits and Practical Applications

The automated platform delivered:

• Substantial time and cost reductions through reduced manual setup and experiential iterations.
• Reproducible method development outcomes aligned with AQbD guidelines.
• Enhanced throughput for multi-component drug analysis and impurity profiling in QC and R&D environments.

Future Trends and Potential Applications

Advances likely to shape automated HPLC method development include:

• Integration with real-time process analytical technologies (PAT) for continuous monitoring.
• Application of machine learning and AI to predict optimal chromatographic conditions.
• Miniaturized and high-throughput HPLC platforms for screening large drug libraries.
• Green chemistry approaches using alternative, sustainable solvents.

Conclusion

The demonstrated automated workflow efficiently developed and validated an HPLC method for Triumeq’s active ingredients and impurities, meeting regulatory resolution and symmetry criteria while drastically reducing development time. This approach exemplifies the benefits of AQbD-driven automation in modern pharmaceutical analysis.

Reference

1. Thermo Fisher Scientific. Application Note 000582: HPLC-DAD method for abacavir, lamivudine, dolutegravir impurities in Triumeq.
2. ViiV Healthcare Press Release: FDA approval for Triumeq.
3. Belk D. Pharma’s 50 Best Sellers. True Cost of Health-Care.
4. Tol T. et al. Simultaneous determination of related substances by HPLC using AQbD. J Chromatogr A, 1432, 26–38.