Analysis of Artesunate and Dihydroartemisinin Using a Core Enhanced Technology Accucore HPLC Column

Significance of the Topic

Artesunate and its active metabolite dihydroartemisinin are key antimalarial agents endorsed by the World Health Organization for treating severe and drug-resistant malaria. Rapid, sensitive quantification of these compounds is essential for pharmaceutical quality control and therapeutic monitoring. Conventional HPLC methods rely on non-MS-compatible buffers, which limit detection sensitivity and throughput. This study demonstrates a fast, MS-friendly analytical approach using Core Enhanced Technology columns.

Objectives and Study Overview

The aim of this application note is to:

• Develop a rapid HPLC method for simultaneous determination of artesunate and dihydroartemisinin with an MS-compatible mobile phase.
• Assess the performance of a 2.6 µm Accucore RP-MS column in terms of analysis speed, peak quality, and method precision.

Methodology

Sample preparation involved dissolving artesunate (1 mg/mL) and α/β-dihydroartemisinin isomers (2 mg/mL) in acetonitrile. The mobile phase consisted of water + 0.1% formic acid and acetonitrile + 0.1% formic acid (60:40 v/v). Separation was performed at 30 °C on a 50 × 2.1 mm Accucore RP-MS column (2.6 µm) with a flow rate of 0.6 mL/min and a 1 µL injection. UV detection was set at 210 nm, with mass spectrometry compatibility.

Used Instrumentation

• HPLC System: Thermo Fisher Scientific Accela 600
• Column: Accucore RP-MS, 2.6 µm superficially porous particles, 50 × 2.1 mm
• Vials: NSC Mass Spec Certified 2 mL glass vials with PTFE-silicone caps
• Reagents: Fisher Scientific HPLC-grade water and acetonitrile; formic acid (0.1%)

Results and Discussion

The method achieved baseline separation of artesunate and α/β-dihydroartemisinin in under 2 minutes. Peak shapes were sharp with minimal tailing. Precision studies (n = 6) yielded retention time %RSD values below 0.5% for all analytes, confirming excellent reproducibility. Low UV response for artesunate underscores the advantage of MS detection for trace-level quantification.

Benefits and Practical Applications

• High throughput: analysis cycle under 2 minutes supports large sample batches.
• MS compatibility: elimination of phosphate buffers enables direct LC-MS coupling.
• Robust performance: Core Enhanced Technology reduces backpressure and secondary interactions, extending column lifespan.

Future Trends and Potential Applications

Advances are expected in the development of sub-2 µm superficially porous particles for ultrafast UHPLC-MS workflows, broader applications in clinical pharmacokinetics and therapeutic drug monitoring, and integration with high-resolution mass spectrometry for comprehensive metabolite profiling.

Conclusion

The Accucore RP-MS column combined with an MS-compatible mobile phase delivers a rapid, precise, and sensitive method for artesunate and dihydroartemisinin analysis. This approach streamlines QC procedures and meets the demands of modern antimalarial drug testing.

References

1. World Health Organization. Artesunate: Final Text for Revision of The International Pharmacopoeia. QAS/09.340/Final, December 2009.
2. American Journal of Tropical Medicine and Hygiene, 58(3):365–368, 1998.