Abstract
We developed a cryo-HPLC/UV method for the simultaneous determination of artemisinin ( 1), α-dihydroartemisinin ( 2α), β-dihydroartemisinin ( 2β), and a ubiquitous thermal decomposition product of 2 (designated as diketoaldehyde, 3), starting from the International Pharmacopoeia monograph on dihydroartemisinin. The method takes for the first time the on-column epimerization process of 2 into consideration. Chromatographic separation was obtained under reversed-phase conditions on a Symmetry C18 column (3.5 μm particle size) with a mobile phase consisting of acetonitrile–water 60:40 (v/v), delivered at 0.60–1.00 ml/min flow-rates, with ultraviolet detection at low wavelength ( λ = 210 nm). Low temperatures ( T = 0–10 °C) were selected on the grounds of a diastereoselective dynamic HPLC (DHPLC) study performed at different temperatures, aimed at identifying the best experimental conditions capable of minimizing the on-column interconversion process.
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