This manuscript reports, the development and validation of a sensitive and selective assay method for simultaneous determination of alpha,beta-arteether and its metabolite dihydroartemisinin (DHA) in rat plasma by liquid chromatography-mass spectrometry. Chromatographic separations were achieved by gradient elution of the analytes with an initial composition of methanol-potassium acetate buffer (pH 4; 73:27, v/v) to 100% methanol in 3 min and maintained for 5 min on a Spheri-10, RP(18) (100 x 4.6 mm i.d.) column following an RP(18) (30 x 4.6 mm i.d.) guard column. The total effluent from the column was split so that one-tenth was injected into the electrospray LC/MS interface. ESI-MS analysis was performed using a Micromass Quattro II Triple Quadrupole Mass Spectrometer equipped with an electrospray source. The MS analysis was carried out at cone voltage of 22 V with a scan range of 200-500 Da. The analytes were quantified from the [M+ K](+) ion chromatograms of alpha,beta-arteether at m/z 352, DHA at m/z 323, artemisinin at m/z 321 and propyl ether analogue of arteether at m/z 365. Liquid-liquid extractions with a combination of n-hexane and hexane-ethyl acetate (8:2) were used to isolate alpha,beta-arteether and DHA from rat plasma. The method was validated and gave good accuracy and precision for the studied domain. Linearity in serum was observed over the range 4.375-70 ng/mL for alpha-arteether and 10-160 ng/mL for beta-arteether and DHA. Percentage bias (accuracy) and within- and between-assay precision were well within the acceptable range. This method was applied to study the pharmacokinetics following oral administration of alpha,beta-arteether (30 mg/kg) in rats.
Copyright 2003 John Wiley & Sons, Ltd.