title: Simultaneous determination of primaquine and carboxyprimaquine in plasma using solid phase extraction and LC-MS assay.
abstract: Sensitive bioanalytical methods are required for pharmacokinetic studies in children, due to the small volume and modest number of samples that can be obtained. We sought to develop a LC-MS assay for primaquine and its active metabolite, carboxyprimaquine, following simultaneous, solid phase extraction of both analytes from human plasma. The analysis was conducted on a single-quad LC-MS system (Shimadzu Model 2020) in ESI+ mode, with quantitation by selected ion monitoring. Primaquine, carboxyprimaquine and 8-aminoquinoline (internal standard) were separated using a mobile phase of 80:20 methanol:water with 0.1% (v/v) formic acid and a Luna C(18) HPLC column, at ambient temperature. Solid phase extraction of the analytes from plasma (0.5 mL) was achieved with Oasis(®) HLB cartridges. The retention times for primaquine, 8-aminoquinoline and carboxyprimaquine were 3.3, 5.7 and 8.5 min, respectively. The calibration curve range (2-1500 μg/L) was appropriate for the limits of quantification and detection for primaquine (2 μg/L and 1μ g/L, respectively) and carboxyprimaquine (2.5 μg/L and 1 μg/L) and the anticipated plasma concentrations of the analytes. Intra- and inter-day precision for both primaquine and carboxyprimaquine was <10% across the concentration range 5-1000 μg/L. Accuracy for both analytes was <15% (5-500 μg/L). This validated LC-MS method with solid phase extraction facilitates the simultaneous analysis of primaquine and carboxyprimaquine from small volumes of human plasma, with run time <10 min, recovery >85% and sensitivity of 1-2 μg/L.
authors: School of Pharmacy, Curtin University, Bentley, Western Australia, Australia.
authors: Madhu
authors: M
authors: Page-Sharp
authors: Kenneth F
authors: KF
authors: Ilett
authors: Inoni
authors: I
authors: Betuela
authors: Timothy M E
authors: TM
authors: Davis
authors: Kevin T
authors: KT
authors: Batty
mesh_terms: D002648:Child; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D011319:Primaquine / Q000031:analogs & derivatives* / Q000097:blood* / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: C035150:8-(3-carboxy-1-methylpropylamino)-6-methoxyquinoline; D011319:Primaquine