Thermo Scientific Pierce FDAA (Marfey's Reagent) reacts with primary amines to enable the separation and quantitation of optical isomers of amino acids by reverse-phase chromatography quick and easy.
FDAA is 1-fluoro-2-4-dinitrophenyl-5-L-alanine amide, also called Marfey's reagent. The compound reacts with primary amines and is used as a derivatization reagent for UV detection in liquid chromatography methods. The reagent provides for detection at 340nm with nanomole sensitivity. FDAA derivatives of D-amino acids exhibit strong intramolecular bonding, which reduces their polarity relative to the corresponding L-amino acid derivatives. Consequently, the D-derivatives are selectively retained on reverse phase columns and elute much later than corresponding L-derivatives.
Highlights:
- For precolumn derivatization of primary amines for detection by UV absorbance (λmax = 340nm)
- Provides nanomole sensitivity in typical HPLC applications with amino acids
- Reaction products enable chiral separation of D- and L-amino acids
- Following reverse-phase HPLC, derivatives have an extinction coefficient of 3 x 10^4M-1cm-1
Reagent Properties:
- Alternative names: FDAA, Marfey's Reagent
- Chemical name: 1-fluoro-2-4-dinitrophenyl-5-L-alanine amide; N2-(5-Fluoro-2,4-dinitrophenyl)-L-alaninamide
- Reactive toward: Primary amines (–NH2)
- Chemical formula: C9H9FN4O5
- CAS number: 95713-52-3
- Molecular weight: 272.19
- Form: Bright yellow or orange crystals or powder, free of particulate matter
Product Details:
FDAA derivatives can be separated with simple linear gradient of triethylamine phosphate/acetonitrile on a Spheri-5 RP-18 HPLC cartridge column. FDAA reduces the need for a chiral column because the D-derivatives are retained on the column, eluting after the L-derivatives. The derivatization is easy to perform and completed in 90 minutes, resulting in derivatives that are stable for at least 48 hours.
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Chemical structure of FDAA (Marfey's Reagent). |
General References:
- Marfey, P., et al. (1984). Carlsberg Res. Comm. 49, 585-590.
- Marfey, P., et al. (1984). Carlsberg Res. Comm. 49, 591-596.
- Szókán, et al. (1988). Applications of Marfey's Reagent in racemization studies of amino acids and peptides. J. Chrom. 444, 115-122.
- Aberhart, D.J., et al. (1985). Separation by high-performance liquid chromato-graphy of (3R)- and 3(S)-beta-Leucine as diastereomeric derivatives. 151, 88-91.
- Martinez del Pozo, et al. (1989). Stereospecificity of reactions catalyzed by bacterial D-amino transaminase. J. Biol. Chem. 264(30), 17784-17789.
- Bhushan, R. and Bruckner, H. (2004). Marfey's reagent for chiral amino acid analysis: A review. Amino Acids 27:231-47.
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