Analysis of underivatized artemisinin and related sesquiterpene lactones by high‐performance liquid chromatography with ultraviolet detection

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Abstract
Introduction: Although high‐performance liquid chromatography with ultraviolet detection (HPLC‐PAD) is widely available, it has not been used for artemisinin (1) analysis because of the lack of UV absorption reported for this lactone. Increased Artemisia annua cultivation for production of 1 requires an affordable and reliable method to analyse 1 and its precursors dihydroartemisinic acid (2) and artemisinic acid (3) simultaneously from underivatized plant extracts.Objective: To validate HPLC‐PAD for the quantification of underivatized artemisinin from A. annua and artemisinin‐based drugs.Methodology: Dried A. annua leaves were extracted with petroleum ether, dried, reconstituted in acetonitrile, and analysed by HPLC‐PAD at 192 nm using an isocratic mobile phase (60:40, acetonitrile:0.1% acetic acid). HPLC‐PAD was evaluated through accuracy, precision, recovery and comparison with HPLC with evaporative light scattering detection (HPLC‐ELSD).Results: HPLC‐PAD proved accurate, precise and reproducible for the direct quantification of 1 and related compounds, and was more sensitive than ELSD for most of the compounds tested. The limit of quantification of 1–3 from plants was 0.048, 0.024 and 0.008 g/100 g dry weight, respectively. Recoveries were over 98%, with good intra‐ and inter‐day repeatability. HPLC‐PAD correlated significantly (r2 = 0.99, p <0.001) with HPLC‐ELSD for artemisinin analysis. HPLC‐PAD was also reliable for the analysis of dihydroartemisinin, artesunate and artelinic acid.Conclusion: HPLC with ultraviolet detection was validated for the quantification of underivatized 1, 2, and 3 from crude plant samples, and is readily applicable for the quality control of herbals and artemisinin‐related pharmaceutical compounds. Copyright © 2008 John Wiley & Sons, Ltd.

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