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Determination of Nicotinic Acid and Nicotinamide in Human Serum by High-Peformance Liquid Chromatography with Postcolumn Ultraviolet-Irradiation and Fluorescence Detection

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Abstract

A high performance liquid chromatographic method involving postcolumn ultraviolet (UV) irradiation and fluorometric detection has been developed for the simultaneous determination of nicotinic acid and nicotinamide in human serum. These compounds are separated by reversed-phase chromatography using a 0.07 M potassium dihydrogen-phosphate solution (pH 4.5) containing 75 mM hydrogen peroxide and 5 uM copper(II) sulfate as a mobile phase. The compounds in the column effluent are irradiated with ultraviolet light to produce fluorescence. This fluorescence is monitored with excitation at 322 nm and emission at 380 nm. The calibration curves for nicotinamide, nicotinic acid and N1-methylnicotinamide are linear over the ranges of 0.6-136, 0.7-166 and 5.5-220 ng, respectively. The mean recoveries of nicotinamide and nicotinic acid from serum are more than 93%.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa, 199-01, Japan

    Ken-ichi Mawatari, Fumio Iinuma & Mitsuo Watanabe

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  1. Ken-ichi Mawatari
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  2. Fumio Iinuma
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  3. Mitsuo Watanabe
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Mawatari, Ki., Iinuma, F. & Watanabe, M. Determination of Nicotinic Acid and Nicotinamide in Human Serum by High-Peformance Liquid Chromatography with Postcolumn Ultraviolet-Irradiation and Fluorescence Detection. ANAL. SCI. 7, 733–736 (1991). https://doi.org/10.2116/analsci.7.733

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  • DOI: https://doi.org/10.2116/analsci.7.733

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