Analysis of porphyrin carboxylic acids in biological fluids by high-performance liquid chromatography |
| |
Authors: | Momoko Chiba Shigeru Sassa |
| |
Institution: | The Rockefeller University Hospital, 1230 York Avenue, New York, New York 10021 USA |
| |
Abstract: | A method for the rapid and sensitive fluorometric analysis of porphyrin carboxylic acids by reverse-phase high-performance liquid chromatography is described. Separation of free porphyrin carboxylic acids was carried out with a microparticulate octadecylsilane column with elution by a gradient of methanol in phosphate buffer containing tetrabutylammonium hydroxide. Separation and quantitation of di-, tri-, tetra-, penta-, hexa-, hepta-, and octacar-boxylic porphyrins was achieved within 25 min at picomolar concentrations. The method is also capable of separating the type I and type III isomers of tetracarboxylic through hexacarboxylic porphyrins. By using a stopped flow technique, one can record fluorescence excitation and emission spectra of porphyrin carboxylic acids. This method is directly applicable to biological fluids such as urine, plasma, red cell lysates, or medium or extracts from cell culture. |
| |
Keywords: | To whom reprint requests should be addressed |
本文献已被 ScienceDirect 等数据库收录! |
|