Fluorescamine reagents

In the first step tin(Il) chloride in acetic acid solution reduces the aromatic nitro groups to amino groups. The aromatic amines produced then react with fluorescamine in weakly basic medium to yield fluorescent derivatives (cf. reagent monograph Fluorescamine Reagent , Volume la). 

Fluorescamine Reagent 297 length UV light (A = 365 nm) these zones could be quantitatively determined after... 

Alkaline hydrolysis was carried out in order to remove PS and PE that can be extracted together with the psychosine. After the alkaline hydrolysis, the extract was washed three times with upper phase containing ammonium hydroxide. If the extract (checked by TLC) had still some contaminants as cerebrosides or sulphatides, a preparative TLC was carried out using chloroform methanol 2 N-NH3 60 35 8 as solvent in order to remove them. The spots were identified by spraying the plate with fluorescamine reagent in acetone (4-phenyl-spiro fu-ran-2 (3H) I -phtalan 3,3-dione) Hoffmann La Roche, Nut ley, U.S.A. 

Sherma, J., and Marzoni, G. (1974). Detection and quantitation of anilines by thin layer chromatography using fluorescamine reagent. Am. Lab. 6(10) 21-30. 

Sherma, J., and Touchstone, J. C. (1974). Quantitative thin layer chromatography of amino acids using fluorescamine reagent. Anal. Lett. 7 279-287. 

Sulfonamides are used for the prevention and treatment of human infectious diseases and also as growth promoters in swine. It is necessary to monitor pork products for drug residues of sulfonamides. Various sulfonamides were separated on Whatman LKCigF layers in a methanol-0.5 M NaCl (50 50) mobile phase spots were detected with UV light (366 nm) after spraying with fluorescamine reagent followed by 0.5% triethanolamine in chloroform to stabilize fluorescence. The Rp values of the sulfonamides were as follows sulfabromomethazine, 0.19 sulfadimethoxine, 0.39 sulfamethazine, 0.59 sulfathiazole, 0.76 sulfaguanidine, 0.90 (Whatman TLC Technical Series, Volume 3). 

Coloration TFG-Start of Zones I C] ninhydrine quench- fluorescamine- reagent ing reagent " )... 

Boc-P-AIa-Try-Met-Asp-Phe Chloroform-methanol-acetic acid (18 2 1) sec-butanol-3% ammonia (25 11 and 3 1) ethanol-acetic acid-water (20 6 11) n-butanol-acetic acid-water (4 1 5, upper phase) Ninhydrin. Ehrlich s reagent, fluorescamine reagent 63... 

Fluorescamine was developed by Weigele et al. in 1972 [8], based on the fact that strongly fluorescent pyrrolinones were formed by the reaction of ninhydrin, phenylacetaldehyde, and primary amines. The reagent, 4-phenylspiro[furan-2(3H),l -phthalan]-3,3 -dione (fluorescamine), is nonfluorescent, and it reacts with primary amines, amino acids, and peptides under aqueous conditions in a few minutes at room temperature to form intensely fluorescent substances (Figure 6.1). On the other hand, nonfluorescent derivatives are formed by the reaction of fluorescamine and secondary amino compoimds. Therefore, fluorescamine can be used for the selective determination of primary amino compounds, and the fluorophore produced by the reaction is the expected pyrrolinone. Because the reaction is sufficiently rapid and the hydrolysis products are nonfluorescent, the fluorescamine reaction is applicable for the postcolumn fluorescence derivatization of primary amino compounds [9]. The amino acids are separated by a cation-exchange column similar to the ninhydrin method, and the column effluent is mixed with an alkaline-buffered solution and fluorescamine reagent. The fluorescent derivatives are detected at 480 nm with excitation at 390 nm. 

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