An HPLC-MS method for simultaneous estimation of alpha,beta-arteether and its metabolite dihydroartemisinin, in rat plasma for application to pharmacokinetic study.
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.
Department of Pharmacokinetics and Metabolism, Central Drug Research Institute, Lucknow-226001, India.
M
M
Rajanikanth
K P
KP
Madhusudanan
R C
RC
Gupta
D000284:Administration, Oral; D000818:Animals; D037621:Artemisinins / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D037621:Artemisinins; C055141:artemotil