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Liquid chromatographic fluorescence determination of amino acids in plasma and urine after derivatization with phanquinone

✍ Scribed by Rita Gatti; Maria Grazia Gioia

Publisher
John Wiley and Sons
Year
2008
Tongue
English
Weight
268 KB
Volume
22
Category
Article
ISSN
0269-3879

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✦ Synopsis

Abstract

Phanquinone (4,7‐phenanthroline‐5,6‐dione) has been investigated as a pre‐column derivatization fluorogenic reagent for liquid chromatographic determination of primary amino acids in biological samples. The derivatization reaction was carried out at 68°C both in the presence of aqueous phosphate buffer (pH 8) for 30 min and without buffer for 60 min to allow the determination of basic amino acids (Orn, Lys, Arg). The resulting derivatives were separated under reversed‐phase HPLC and detected at λ~em~ = 460 nm with λ~ex~ = 400 nm. The proposed method was validated and applied to the determination of a variety of amino acids directly in urine and after deproteinization with 5‐sulfosalicylic acid in plasma samples. The detection and quantitation limits were found in the range 10–450 and 35–1400 fmol, respectively__.__ Copyright © 2007 John Wiley & Sons, Ltd.

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