Alternative method for determination of pyrimethamine in plasma by high-performance liquid chromatography |
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Authors: | K Na-Bangchang P Tan-ariya R Ubalee B Kamanikom J Karbwang |
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Institution: | aClinical Pharmacology Unit, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Pyathai, Bangkok 10400, Thailand;bDepartment of Microbiology, Faculty of Science, Mahidol University, 420/6 Rajvithi Road, Pyathai, Bangkok 10400, Thailand |
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Abstract: | A rapid, selective, sensitive and reproducible reversed-phase high-performance liquid chromatography (HPLC) procedure for the quantitative determination of pyrimethamine (PYR) in plasma is described. The procedure involved the two-step extraction of PYR and the internal standard, quinine (QN) with acetonitrile and dichloromethane at basic pH. Chromatographic separation consisted of the mobile phase (methanol-water containing 0.005 M octanesulfonic acid, 50:50, v/v) running through the column (Techopak-10 C18) at a flow-rate of 1.6 ml/min. Detection was at UV wavelength of 240 nm. The mean recoveries of PYR and QN at a concentration range of 50 and 500 ng/ml were 98.9 and 89%, and 94.7 and 96% for PYR and QN. The within-day coefficients of variation were 2.1–5.1% for PYR and 5.9% for QN. The day-to-day coefficients of variation were 2.1–4.1% for PYR and 5% for QN. The minimum detectable concentrations for PYR and QN in plasma were 3 and 10 ng/ml. The method was found to be suitable for use in clinical pharmacokinetic study. |
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Keywords: | Pyrimethamine |
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