NBD-F

- Overview
  - NBD-F is highly reactive and can label primary and secondary amines under mild conditions (1 min reaction at 60°C in a weak basic solution). NBD-F is a pre-labeling compound for HPLC analysis of small molecules. NBD-labeled compounds are orange with a maximum wavelength at 470 nm.The excitation and emission of the derivatized compound are 470 nm and 530 nm, respectively.
    
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- Properties
  - Categories
    
    HPLC Derivertization Reagents
    
    CAS Number
    
    29270-56-2
    
    Molecular Formula
    
    C6H2FN3O3
    
    Molecular Weight
    
    183.1
    
    Appearance
    
    Pale yellow powder
    
    Purity
    
    ≥99.0% (HPLC)
    
    Storage
    
    -20°C, protect form light and moisture
    
    Shipping
    
    ambient temperature
    
    * For research use only
- Applications
  - Application Description
    
    NBD Labeling Protocol
    1. To prepare sample solution, mix or dissolve a sample with 50 mM borate buffer (pH 8.0) containing 20 mM EDTA.
    2. Mix 300 μl of the sample solution and 100 μl of 100 mM NBD-F/acetonitrile solution in a reaction vial.
    3. Heat the vial at 60°C for 1 minute and then cool it on an ice bath.
    4. Add 400 μl of 50 mM HCl aqueous solution to the reaction mixture.
    5. Use this mixture for HPLC analysis to determine NBD-labeled compounds.
- Reference
  - 1) K. Imai, et al., Fluorimetric Determination of Secondary Amino Acids by 7-fluoro-4-nitrobenzo-2-oxa-1, 3-diazole. Anal Chim Acta. 1981;130:377-383.
    
    2) Y. Watanabe, et al., High-Performance Liquid Chromatography and Sensitive Detection of Amino Acids Derivatized with 7-fluoro-4-nitrobenzo-2-oxa-1, 3-diazole. Anal Biochem. 1981;116:471-472.
    
    3) Y. Watanabe, et al., Pre-column Labelling for High-performance Liquid Chromatography of Amino Acids with 7-fluoro-4-nitrobenzo-2-oxa-1, 3-diazoleand Its Application to Protein Hydrolysates. J Chromatogr A. 1982;239:723-732.
    
    4) T. Toyo’oka, et al., Reaction of Amines of Biological Importance with 4-fluoro-7-nitrobenzo-2-oxa-1, 3-diazole. Anal Chim Acta. 1983;149:305-312.
    
    5) Y. Watanabe, et al., Liquid Chromatographic Determination of Amino and Imino Acids and Thiols by Postcolumn Derivatization with 4-Fluoro-7-nitrobenzo-2, 1, 3-oxadiazole. Anal Chem. 1983;55:1786-1791.
    
    6) Y. Watanabe, et al., Sensitive Detection of Amino Acids in Human Serum and Dried Blood Disc of 3mm Diameter for Diagnosis of Inbornerrors of Metabolism. J Chromatogr. 1984;309:279-286.
    
    7) H. Miyano, et al., Further Studies on the Reaction of Amines and Proteins with 4-fluoro-7-nitrobenzo-2-oxa-1, 3-diazole. Anal Chim Acta. 1985;170:81-87.
    
    8) H. Kotaniguchi, et al., Automatic Amino Acid Analysis Utilizing 4-fluoro-7-nitrobenzo-2-oxa-1, 3-diazole. J Chromatogr. 1987;420:141-145.
    
    9) K. Imai, et al., High-Performance Liquid Chromatography with Photochemical Fluorimetric Detection of Tryptophan Based on 4-fluoro-7-nitrobenzo-2-oxa-1, 3-diazole Total Protein Amino Acid Analysis. Anal Chim Acta. 1988;205:7-14.
    
    10) K. Imai, et al., Dynamic Analytical Chemistry: A Trial Study of the Interaction of Fluorogenic Reagents with Living Chinese Hamster Ovary Cells. Anal Chim Acta. 1989;223:299-308.
    
    11) C. Nakamura, et al., Dioxin-binding Pentapeptide for Use in a High-sensitivity On-bead Detection Assay. Anal Chem. 2005;77:7750-7757.

MSDS

NBD-F

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