Chemical Properties
yellow to light brown crystalline powder
Uses
4-Chloro-7-nitrobenzofurazan is used as a derivatizing reagent for chromatography analysis of amino acids and low molecular weight amines. It is utilized in the preparation of fluorescent phospholipid-derivative, hydroxynaphthofurazan and 4-chloro-7-nitrobenzofurazan- didecanoylphosphatidylethanolamine. It serves as a fluorescent reagent to label free sulfhydryls and N-terminals within proteins and trapping agent for cysteine sulfenic acid. It is a sensitive chromogenic and fluorogenic reagent. It reacts with trans-1-methoxy-3-(trimethylsilyloxy)-1,3-butadiene (Danishefsky's diene) to prepare regioselectively the silyl enol ether. Further, it is used to label peptides, proteins and other biomolecules. In addition it is used in the synthesis of fluorescent phospholipid-derivative, NBD-didecanoylphosphatidylethanolamine, functionalized hydroxynaphthofurazan and 7-nitrobenzofurazan (NBD)-labeled maleimide.
Uses
4-Chloro-7-nitrobenzofurazan is nonfluorescent until it reacts with primary or secondary amines to produce a fluorescent product. 4-Chloro-7-nitrobenzofurazan has been extensively used as a derivatizing reagent for chromatography analysis of amino acids and low molecular weight amines.
Definition
ChEBI: A benzoxadiazole that is 2,1,3-benzoxadiazole which is substituted at position 4 by chlorine and at position 7 by a nitro group.
General Description
4-Chloro-7-nitrobenzofurazan (NBD-Cl) is a highly sensitive chromogenic and fluorogenic reagent. It is reported to react spontaneously with
trans-1-methoxy-3-(trimethylsilyloxy)-1,3-butadiene (Danishefsky′s diene), to afford regioselectively the silyl enol ether, via normal electron-demand Diels-Alder (NEDDA) reaction. A study of the molecular structure, vibrational spectra and NBO (Natural Bond Orbital) analysis of NBD–Cl has been undertaken. Its utility as a pre-column derivatization agent has been examined.
in vitro
4-Chloro-7-nitrobenzofurazan(NBD chloride) forms highly fluorescent derivatives for detection of all the protein amino acids. In addition, NBD-Cl provides a simple and sensitive method for determination of N-terminal amino acids. The differences in intensity and color of fluorescence can be used to advantage to identify prolyl peptides.
Purification Methods
Wash the solid with H2O, and it recrystallises from aqueous EtOH (1:1) as pale yellow needles. It sublimes in a vacuum [Gosh & Whitehouse Biochem J 108 155 1968, UV, NMR: Bolton et al. J Chem Soc 1004 1966].
