高效液相色谱法测定血浆中乙胺嘧啶的替代方法。
Alternative method for determination of pyrimethamine in plasma by high-performance liquid chromatography.
作者信息
Na-Bangchang K, Tan-ariya P, Ubalee R, Kamanikom B, Karbwang J
机构信息
Clinical Pharmacology Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
出版信息
J Chromatogr B Biomed Sci Appl. 1997 Feb 21;689(2):433-7. doi: 10.1016/s0378-4347(96)00327-1.
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.
本文描述了一种快速、选择性好、灵敏且可重现的反相高效液相色谱法（HPLC），用于定量测定血浆中的乙胺嘧啶（PYR）。该方法包括在碱性pH条件下，用乙腈和二氯甲烷对PYR和内标物奎宁（QN）进行两步萃取。色谱分离采用流动相（含0.005 M辛烷磺酸的甲醇 - 水，50:50，v/v）以1.6 ml/min的流速通过色谱柱（Techopak - 10 C18）。检测波长为240 nm的紫外光。在50和500 ng/ml的浓度范围内，PYR和QN的平均回收率分别为98.9%和89%，以及94.7%和96%。PYR的日内变异系数为2.1 - 5.1%，QN为5.9%。PYR的日间变异系数为2.1 - 4.1%，QN为5%。血浆中PYR和QN的最低检测浓度分别为3和10 ng/ml。该方法适用于临床药代动力学研究。
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