Quantification of artemisinin in human plasma using liquid chromatography coupled to tandem mass spectrometry |
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Authors: | N. Lindegardh J. Tarning P.V. Toi T.T. Hien J. Farrar P. Singhasivanon N.J. White M. Ashton N.P.J. Day |
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Affiliation: | 1. Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand;2. Nuffield Department of Clinical Medicine, Centre for Tropical Medicine, University of Oxford, Oxford, UK;3. Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam;4. Department of Pharmacology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden |
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Abstract: | A liquid chromatographic tandem mass spectroscopy method for the quantification of artemisinin in human heparinised plasma has been developed and validated. The method uses Oasis HLB™ μ-elution solid phase extraction 96-well plates to facilitate a high throughput of 192 samples a day. Artesunate (internal standard) in a plasma–water solution was added to plasma (50 μL) before solid phase extraction. Artemisinin and its internal standard artesunate were analysed by liquid chromatography and MS/MS detection on a Hypersil Gold C18 (100 mm × 2.1 mm, 5 μm) column using a mobile phase containing acetonitrile–ammonium acetate 10 mM pH 3.5 (50:50, v/v) at a flow rate of 0.5 mL/min. The method has been validated according to published FDA guidelines and showed excellent performance. The within-day, between-day and total precisions expressed as R.S.D., were lower than 8% at all tested quality control levels including the upper and lower limit of quantification. The limit of detection was 0.257 ng/mL for artemisinin and the calibration range was 1.03–762 ng/mL using 50 μL plasma. The method was free from matrix effects as demonstrated both graphically and quantitatively. |
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Keywords: | Antimalarial Artemisinin High throughput Liquid chromatography/tandem mass spectrometry (LC/MS/MS) Solid phase extraction (SPE) |
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