Quantification of artemisinin in human plasma using liquid chromatography coupled to tandem mass spectrometry.
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 mu-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 microL) 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 x 2.1 mm, 5 microm) column using a mobile phase containing acetonitrile-ammonium acetate 10mM 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 microL plasma. The method was free from matrix effects as demonstrated both graphically and quantitatively.
Lindegardh
N
N
Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand. niklas@tropmedres.ac
Tarning
J
J
Toi
P V
PV
Hien
T T
TT
Farrar
J
J
Singhasivanon
P
P
White
N J
NJ
Ashton
M
M
Day
N P J
NP
D000704:Analysis of Variance; D000962:Antimalarials / Q000097:blood* / Q000493:pharmacokinetics; D037621:Artemisinins / Q000097:blood* / Q000493:pharmacokinetics; D000077332:Artesunate; D001682:Biological Availability; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007202:Indicators and Reagents; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D000962:Antimalarials; D037621:Artemisinins; D007202:Indicators and Reagents; D000077332:Artesunate; C031327:artemisinin