PRECOLUMN FLUORESCENCE DERIVATIZATION OF CARNITINE AND ACYLCARNITINESWITH 4-(2-AMINOETHYLAMINO)-7-NITRO-2,1,3-BENZOXADIAZOLE PRIOR TO HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
K. Matsumoto et al., PRECOLUMN FLUORESCENCE DERIVATIZATION OF CARNITINE AND ACYLCARNITINESWITH 4-(2-AMINOETHYLAMINO)-7-NITRO-2,1,3-BENZOXADIAZOLE PRIOR TO HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, Journal of chromatography, 678(2), 1994, pp. 241-247
4-(2-Aminoethylamino)-7-nitro-2,1,3-benzoxadiazole (NBD-ED) was synthe
sized as a precolumn fluorescence derivatization reagent for the high-
performance liquid chromatographic determination of carnitine and acyl
carnitines. Carnitine and acylcarnitines were reacted with NBD-ED (2.0
mM) and 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (35 mM) in pyr
idine-dimethylformamide (1:4) at room temperature for 2 h. The NBD-ED
derivatives of carnitine and acylcarnitines were separated by gradient
elution with water-acetonitrile containing 10 mM trifluoroacetic acid
on a reversed-phase column and the eluate was monitored with excitati
on at 470-485 nm and emission at 530-540 nm. Carnitine and sixteen acy
lcarnitines were determined within 45 min. The detection limits (signa
l-to-noise ratio=3) for the carnitine-related compounds were 10-100 fm
ol. Four other NBD-alkyldiamines were also studied.