DETERMINATION OF CHLORPHENIRAMINE IN HUMAN BLOOD SERUM

扑热息痛中低百万分之几的测定 正在开发血清以支持即将进行的一项临床研究 在保罗·图克尔陶布的指导下进行，医学博士，实验室 免疫生物化学、DAPP、CBER。有一种程序，已经过评估 以及部分验证，包括提取、浓缩和气体 层析法。选择性离子监测法检测扑尔敏 M/e 203处特征碎片离子的质谱学。 萃取效率和进样量通过添加 四氢扑尔敏及其类似物D4的监测 碎片离子的m/e为207。开发一种替代方法，那将是 允许扑尔敏拆分为对映体 组件，正在进行中。对映体拆分尚未完成 用气相色谱手性柱得到。高效液相色谱柱 在α-糖蛋白和β-环糊精固定相上 提供了对映体拆分，但在 Ppb水平不是通过紫外线吸光度来实现的。其他高效液相色谱 检测方法尚未被评估为提供必要的 敏感度。柱前衍生化方法的工作正在进行中 涉及与氯甲酸苄酯的反应、手性高效液相和荧光 可提供必要的灵敏度以检测 扑尔敏在血清平衡水平，同时允许对映体 物种形成。

The determination of chlorpheniramine at low part per billion levels in blood serum is being developed to support a clinical study to be conducted under the direction of Paul Turkeltaub, M.D., Lab of Immunobiochemistry, DAPP, CBER. One procedure, that has been evaluated and partially validated, involves extraction, concentration and gas chromatography. Chlorpheniramine is detected by selective ion monitoring mass spectrometry of a characteristic fragment ion at m/e 203. Extraction efficiency and injection volume are normalized by addition of tetradeuterated chlorpheniramine and monitoring of the analogous D4 fragment ion at m/e 207. Development of an alternate method, that would permit the resolution of chlorpheniramine into its enantiometic components, is in progress. Enantiomeric resolution has not been obtained using gas chromatographic chiral columns. HPLC columns based on alpha-glycoprotein and beta-cyclodextrin stationary phases have provided enantiomeric resolution, but detection of chlorpheniramine at ppb levels has not been accomplished by UV absorbance. Other HPLC detection methods have not been evaluated as providing necessary sensitivity. Work is in progress on a pre-column derivatization method involving reaction with benzylchloroformate, chiral HPLC and fluorescence detection that may provide the necessary sensitivity for detection of chlorpheniramine at serum equilibrium levels while allowing enantiomeric speciation.