Lévai F, Liu C M, Tse M M, Lin E T
Pharmacokinetic and Metabolism Laboratory, CHINOIN Pharmaceutical and Chemical Works Co. Ltd., Budapest, Hungary.
Acta Physiol Hung. 1995;83(1):39-46.
A sensitive HPLC assay utilising a simple fluorescence pre-column labelling technique was developed for plasma level monitoring of different types of mono- and dicarboxylic acids. Carboxylic acids form mixed anhydrides with ethyl chloroformate in the presence of triethylamine; the mixed anhydrides further react with L-leucine-4-methyl-7-coumarinylamide, forming highly fluorescent and stable amides. Drugs with no chromophore (azelaic acid, and its longer carbon chain analogues) or week UV absorption (artelinic acid, enalaprilat) were used as model compounds. The plasma samples were extracted using ion exchange solid phase cartridges. The separation was performed on an Axxiom C18 (5 microns, 4.6 x 250 mm) column. The detector wavelengths were set at 330 nm for excitation and 390 nm for emission.
开发了一种灵敏的高效液相色谱法，采用简单的荧光柱前标记技术来监测不同类型一元和二元羧酸的血浆水平。羧酸在三乙胺存在下与氯甲酸乙酯形成混合酸酐；混合酸酐再与L-亮氨酸-4-甲基-7-香豆素酰胺反应，形成高荧光且稳定的酰胺。选用无发色团的药物（壬二酸及其长碳链类似物）或弱紫外吸收的药物（阿替林酸、依那普利拉）作为模型化合物。血浆样本用离子交换固相柱进行萃取。分离在Axxiom C18（5微米，4.6×250毫米）柱上进行。检测器波长设定为激发波长330纳米，发射波长390纳米。
