Johansson B
J Chromatogr. 1986 Aug 22;381(1):107-13. doi: 10.1016/s0378-4347(00)83569-0.
A method for on-line high-performance liquid immunoaffinity chromatographic analysis of 5,5-diphenylhydantoin (phenytoin) in human plasma is described. The technique is simple and does not require sample preparation or addition of an internal standard, and the phenytoin is eluted from the columns in 12 min. A phenytoin-specific polyclonal immunoglobulin was attached to an organic silane derivative on silica. The immunosorbent was packed into a short liquid chromatography column and installed in a modified high-performance liquid chromatography system for on-line sample pre-purification. Standard curve linearity of plasma phenytoin was obtained at concentrations up to 160 mumol/l. When compared to conventional high-performance liquid chromatography, plasma phenytoin levels gave a correlation coefficient of 0.993. The phenytoin is highly bound to plasma proteins (ca. 87%). When plasma is injected onto the pre-column, a dissociation and partition of phenytoin from plasma proteins to a complete association with on-column insolubilized antibodies results.
本文描述了一种用于在线高效液相免疫亲和色谱分析人血浆中5,5-二苯基乙内酰脲（苯妥英）的方法。该技术简单，无需样品制备或添加内标，苯妥英在12分钟内从柱上洗脱。一种苯妥英特异性多克隆免疫球蛋白连接到硅胶上的有机硅烷衍生物上。将免疫吸附剂填充到短液相色谱柱中，并安装在改进的高效液相色谱系统中用于在线样品预纯化。血浆苯妥英的标准曲线在浓度高达160μmol/L时呈线性。与传统高效液相色谱相比，血浆苯妥英水平的相关系数为0.993。苯妥英与血浆蛋白高度结合（约87%）。当将血浆注入预柱时，苯妥英会从血浆蛋白中解离并分配，然后与柱上固定化抗体完全结合。
