Wong L T, Solomonraj G, Thomas B H
J Chromatogr. 1977 May 11;135(1):149-54. doi: 10.1016/s0021-9673(00)86311-6.
An improved high-pressure liquid chromatography method for the estimation of warfarin in plasma was developed. Plasma was acidified and extracted with ethylene dichloride spiked with methylated warfarin [3-(alpha-acetonylbenzyl)-4-methoxy-coumarin] as internal standard. The residue, redissolved in dioxane, was chromatographed on a reversed-phase column using a mobile phase of 40% dioxane in water (pH 4.2) on a high-pressure liquid chromatograph fitted with an UV absorbance detector. Recoveries from extraction, quantitated using tracer amounts of [14C]warfarin and methylated [14C]warfarin were 92.2 +/- 3.16% and 82.33 +/- 1.03%, respectively. The standard curve was linear between o.625 and 5.0 microng/ml. Detection was sensitive to approximately 0.5 microng/ml and specific without the inter ference of normal plasma constituents and warfarin metabolites.
开发了一种改进的高压液相色谱法用于测定血浆中的华法林。将血浆酸化，并用加有甲基化华法林[3-(α-丙酮基苄基)-4-甲氧基香豆素]作为内标的二氯乙烷进行萃取。残渣用二氧六环重新溶解后，在配备有紫外吸收检测器的高压液相色谱仪上，于反相柱上以40%二氧六环的水溶液（pH 4.2）作为流动相进行色谱分析。使用示踪量的[14C]华法林和甲基化[14C]华法林进行定量，萃取回收率分别为92.2±3.16%和82.33±1.03%。标准曲线在0.625至5.0微克/毫升之间呈线性。检测灵敏度约为0.5微克/毫升，且具有特异性，不受正常血浆成分和华法林代谢产物的干扰。
