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Quantitation of agmatine by liquid chromatography with laser-induced fluorescence detection |
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| Authors: | Shulin Zhao Yangzheng Feng Michael H. LeBlanc John E. Piletz Yi-Ming Liu |
| Affiliation: | a Department of Chemistry, Jackson State University, 1400 J.R. Lynch Street, Jackson, MS 39217-0510, USA b Department of Pediatrics, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS 39216, USA c Department of Psychiatry and Human Behavior, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS 39216, USA |
| Abstract: | A high performance liquid chromatography (HPLC) method is described for the determination of agmatine, an endogenous neuromodulator. The method involves pre-column derivatization of the sample with a fluorescent tagging reagent, 7-fluoro-4-nitrobenzoxadiazole (NBD-F). The resulting agmatine derivative is stable and can be readily extracted into ethyl acetate at pH 8.5. The extraction enhances the quantification of low level agmatine because it eliminates chromatographic peaks caused by endogenous amino acids. The HPLC separation is carried out on a C8 reversed phase column and completed in less than 10 min. With laser-induced fluorescence (LIF) detection, the detection limit is 5×10−9 M agmatine. Method precision (coefficient of variation) is 5% for agmatine in human plasma at the sub-μM level. This method has been validated by determination of agmatine in biological samples including human plasma and rat brain and stomach tissues. |
| Keywords: | Agmatine Biological sample HPLC Laser-induced fluorescence detection 7-Fluoro-4-nitrobenzoxadiazole |
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