High-performance liquid chromatographic determination of N1-alkylnicotinamide in urine |
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| Authors: | T Hirayama K Yoshida K Uda M Nohara S Fukui |
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| Affiliation: | 1. School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong, China;2. ICBMS UMR 5246, Universite Lyon 1, CNRS, INSA Lyon, CPE Lyon, Batiment Raulin, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne, France;1. Division of Physical Science and Engineering, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia;2. Clean Combustion Research Center, Division of Physical Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia;3. Division of Computer, Electrical, and Mathematical Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia |
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| Abstract: | A simple high-performance liquid chromatographic method has been developed for determining N1-alkylnicotinamides, including C1-C5 alkyl derivatives, in urine. N1-Alkylnicotinamides were reacted with acetophenone in strong alkali medium at 0 degrees C and then formic acid was added. The reaction mixture was heated in acidic medium at above 93 degrees C, and the fluorescent product, 1-alkyl-7-phenyl-1,5-dihydro-5-oxo-1,6-naphthyridine, was chromatographed by HPLC, using a Zorbax SCX-300 column with a mixed mobile phase of acetonitrile-0.04 M ammonium phosphate, monobasic. N1-Alkylnicotinamides can be determined as 1,6-naphthyridine derivatives by a fluorometric detector at a level of 100 pg (signal/noise = 2). Recoveries of N1-alkylnicotinamides in urine were satisfactory. Interfering reaction products from NAD+ and NADP+ were clearly eliminated for determination of N1-alkylnicotinamides without pentyl derivatives. |
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