DETERMINATION OF CHLORPHENIRAMINE IN HUMAN BLOOD SERUM

目前正在开发血清中低含量扑尔敏的检测方法，以支持在DAPP CBER免疫生物化学实验室的指导下进行的一项临床研究。其中一个程序已经过评估和部分验证，涉及提取、浓缩和气相色谱。扑尔敏的特征碎片离子的选择离子监测质谱图的m/e为203。通过加入四氢扑尔敏和监测类似的D4碎片离子的m/e 207，使萃取效率和进样量归一化。另一种方法的开发正在进行中，这种方法可以将扑尔敏拆分成对映体成分。用气相色谱手性柱还没有得到对映体的拆分。基于α-糖蛋白和β-环糊精固定相的高效液相色谱柱已经提供了对映体的拆分，但在ppb水平上的扑尔敏的检测不是通过紫外吸收来完成的。其他高效液相检测方法还没有被评估为提供了必要的灵敏度。氯甲酸苄酯的柱前衍生化、手性高效液相色谱和荧光检测可在允许对映体形成的同时，为扑尔敏在血清平衡水平上的检测提供必要的灵敏度。

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. A pre- column derivatization method involving reaction with benzylchloroformate, chiral HPLC and fluorescence detection may provide the necessary sensitivity for detection of chlorpheniramine at serum equilibrium levels while allowing enantiomeric speciation.