A rapid method for the determination of artemisinin and its biosynthetic precursors in Artemisia annua L. crude extracts

Abstract

A rapid high-pressure liquid chromatography (HPLC) tandem mass spectrometry (TQD) method for the determination of artemisinin, 9-epi-artemisinin, artemisitene, dihydroartemisinic acid, artemisinic acid and arteannuin B in Artemisia annua extracts is described. Detection and quantification of 9-epi-artemisinin in crude extracts are reported for the first time. In this method all six metabolites are resolved and eluted within 6 min with minimal sample preparation. A recovery of between 96.25% and 103.59% was obtained for all metabolites analysed and the standard curves were linear (r(2)>0.99) over the concentration range of 0.15-10 μg mL(-1) for artemisinin, 9-epi-artemisinin, artemisitene and arteannuin B, and the range of 3.75-120 μg mL(-1) for dihydroartemisinic acid and artemisinic acid. All validation indices were satisfactory, showing the method to be robust, quick, sensitive and adequate for a range of applications including high throughput (HTP) analysis.

Keywords: 9-Epi-artemisinin; Artemisinin; Liquid chromatography; Multiple reaction monitoring (MRM); Tandem mass spectrometry.