NBD-F (=4-Fluoro-7-nitro-2,1,3-benzoxadiazole) [for HPLC Labeling]

The compound 1 which is an HPLC fluorescence labeling reagent having a 2,1,3-benzoxadiazole skeleton, can easily react with amino or mercapto groups to form the corresponding derivatives. 1 itself does not fluoresce, and its ethanol solution is relatively stable for a week in a refrigerator. The derivatives can reach the detector without any decomposition under reversed phase HPLC. An excellent chromatogram can be obtained by fluorescence detection at the excitation and emission wavelengths of 470 nm and 530 nm, respectively. Since their excitation and fluorescence wavelengths are at longer wavelengths, detection has less interference by contaminants. Thus, further highly sensitive detection can be done by using laser induced fluorescence detector. When the reagent is hydrolyzed (NBD-OH), its fluorescence can be erased under an acidic condition. Therefore, this hydrolyzed reagent can be used as a post column reaction reagent^5,7).

Application example:
[Amino acids] ^2,3)
To 10 µL of 50 µM amino acid standard solution, add 10 µL of 0.1 M boric acid buffer solution (pH 8.0) and 20 µL of 50 mM labeling reagent 1 in ethanol solution, and incubate the mixture at 60 °C for 1 min. Immediately cool it with ice bath, and add 460 µL of 5 mM HCl to the reactant solution. Use 10 µL of the solution as an HPLC sample.

TCI Products Pamphlet
HPLC Labeling Reagents ( PDF 10MB )