Thermo Scientific Pierce Marfey's Reagent (FDAA) reacts with primary amines to enable the quick and easy separation and quantitation of optical isomers of amino acids by reverse-phase chromatography.
Features of Marfey's Reagent (FDAA):
? 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-1 cm-1
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.
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
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.
