Sensitive determination of enantiomers of amino acids derivatized with the fluorogenic reagent, 4-fluoro-7-nitro-2,1,3-benzoxadiazole, separated on a Pirkle-type column, Sumichiral OA 2500(S)

Abstract

Some L- and D-amino acids (Phe or Leu) were derivatized with a fluorogenic reagent, 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) and separated on a Pirkle-type column, Sumichiral OA 2500(S) [(S)-1-naphthylglycyl-3,5-dinitrophenylamide silica gel] with a mobile phase of 20 mM ammonium acetate in methanol. The fluorometric detection was made at 530 nm with excitation at 470 nm. No racemization of the enantiomers occurred during the derivatization reaction. The separation factors (alpha) for NBD-L-Phe and NBD-D-Phe, and NBD-L-Leu and NBD-D-Leu, were 1.27 and 1.17, respectively. The detection limits were in the range of ca. 30 fmol.