A fully automated amino acid analyzer using NBD-F as a fluorescent derivatization reagent.
A fully automated amino acid analyzer using NBD-F (4- fluoro-7-nitro-2,1,3-benzoxadiazole) as a fluorescent derivatization reagent was developed. The whole analytical process was fully automated from derivatization, injection to HPLC separation and quantitation. The derivatization reaction conditions were re-evaluated and optimized. Amino acids were derivatized by NBD-F for 40 min at room temperature in the borate buffer (pH 9.5). The derivatives were separated within 100 min and fluorometrically detected at 540 nm with excitation at 470 nm. The detection limits for amino acids were in the range of 2.8-20 fmol. The calibration curves were linear over the range of 20 fmol to 20 pmol on column with the correlation coefficients of 0.999. The coefficients of variation were less than 5% at 3 pmol injection for all amino acids. Amino acids in rat plasma were determined by the proposed HPLC method.
Aoyama
Chiaki
C
The Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Santa
Tomofumi
T
Tsunoda
Makoto
M
Fukushima
Takeshi
T
Kitada
Chieko
C
Imai
Kazuhiro
K
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives* / Q000737:chemistry*; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001331:Automation; D004864:Equipment and Supplies; D005456:Fluorescent Dyes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008297:Male; D051381:Rats; D011921:Rats, Inbred WKY; D012680:Sensitivity and Specificity
D000596:Amino Acids; D005456:Fluorescent Dyes; C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; D009327:4-Chloro-7-nitrobenzofurazan