Dansyl chloride

Treat the sample for 30 to 40 imn with sodium hydroxide solution (c = 1 mol/1) at 80 °C Apply sample solution and then 0 2% dansyl chloride over It Cover with a glass plate and allow to react in the dark at room temperature for 60 mm, then chromatograph... 

An excellent discussion on derivatization techniques has been given by Lawrence (17) including a detailed discussion on pre-column derivatization (18) and post-column derivatization (19). Probably, the more popular procedures are those that produce fluorescing derivatives to improve detector sensitivity. One of the more commonly used reagents is dansyl chloride (20), 5-dimethylamino-naphthalene-1-sulphonyl chloride (sometimes called DNS-chloride or DNS-C1). The reagent reacts with phenols and primary and secondary amines under slightly basic conditions forming sulphonate esters or sulphonamides. 

Reports on the use of fluorescent derivatives abound (5). Some reagents have become widely used. The dansyl group is probably the most thoroughly studied. Dansyl chloride has been widely used as a fluorescent derivatizing reagent for HPLC (6,7). It reacts readily with primary and secondary amino groups (7) and with phenols (8), but forms derivatives of alcohols very slowly (9). The lower detection limit for dansyl derivatives of aliphatic amines is in the range of 300 femtomoles per injection. 

Dansyl chloride does have some drawbacks when used with reverse-phase HPLC. One of these is that the quantum yield of fluorescence varies greatly with the polarity of the solvent for dansyl derivatives (10,11). As the polarity of the solvent increases, the... 

Dipping solution n Dissolve 100 mg dansyl chloride (S-(dimethylamino)-naphtha-lene-l-sulfonyl chloride) in 100 ml ethanol [6]. ... 

Spray solution Dissolve 50 mg dansyl chloride in 100 ml acetone [3-5]. 

Dansyl chloride that exhibits a blue intrinsic fluorescence, reacts with many amines and phenols to yield derivatives with fluorescence of another color. 

Note Tertiary amines do not react with dansyl chloride and can be detected by spraying afterwards with Wagner s reagent [1]. The detection limits for amines are in the lower nanogram range. 

Gray, R. "Sequence Analysis with Dansyl Chloride", In "Methods In Enzymology", p. 333, Vol. XXV, "Enzyme Structure, Part B", C. H. W. Hlrs and S. N. Tlmasheff, Editors, Academic Press, New York, 1972. 

Dansyl chloride Dns-Cl Primary and secondary amines, phenols, euaino acids and imidazoles 550, 617-619... 

Dansyl chloride is the most widely used of the derivatizing reagents. It forms derivatives with primary and secondary amines readily, less rapidly with phenols and imidazoles, and very slowly with alcohols. The reaction medium is usually an aqueous-organic sixture (e.g., 1 1 acetone-water) adjusted to a pH of 9.5-10. Dansyl chloride has two major application areas. It is used to determine small amounts of amines, amino acids and phenols, as... 

The Af-dansylated amino acid (e.g., glycine, leucine, proline) exhibits a yellow fluorescence. The sensitivity of detection for amino acids by this method is about 10 9 mol of amino acid. The advantage of this prodecure in comparison with that using dansyl chloride is the fact that it can be carried out in homogenous aqueous solution without addition of a cosolvent. 191... 

The amine-reactive 5-(dimethylamino)naphthalene-l-sulfonyl (dansyl) chloride 28 [80] and related fluorophores [81, 82], as well as the 5-((2 aminoethyl)amino) naphthalene-1-sulfonic acid (EDANS) 29, are included in the naphthalene fluorophore family. Derivatives of the latter, such as compound 30, exhibit a Lm.ix/ Lem 336/520 nm, molar absorptivity (e) of 6.1 x 103 M-1 cm-1, and a fluorescent quantum yield of 0.27 in water [83], The use of EDANS is particularly interesting in FRET experiments [84, 85]. Furthermore, 4-amino-3,6-disulfonylnaphthalimides (e.g., Lucifer yellow 31), associated to a longer absorption (Lmax 428 nm) [86] are suitable polar tracers [87]. 

El, E2 El, E2, 17ot-E2, EE2, E3 Cleaning SPE and LC SBSE Centrifugation, lyophilization, grinding, sieving USE Cleanup SPE, derivatization with dansyl chloride, LLE with hexane, silica gel column chromatography... 

Amines are another important group of analytes. Mellbin and Smith [72] compared three different fluorescent reagents, dansyl chloride, 4-chloro-7-nitrobenzo-1,2,5-oxadiazole, and o-phthaldialdehyde, for derivatization of alkylamines. The dansyl tag was found to be the most effective. Hamachi et al. [73] described the application of an HPLC-POCL method for determination of a fluorescent derivative of the synthetic peptide ebiratide. Another comparative study was done by Kwakman et al. [74], where naphthalene-2,3-dialdehyde and anthracene-2,3-dial-dehyde were evaluated as precolumn labeling agents for primary amines. The anthracene-2,3-dialdehyde derivatives were not stable, especially in the presence of hydrogen peroxide, and the POCL detection of these derivatives was therefore... 

Table 4 illustrates the use of the CAR technique to develop CL kinetic-based determinations for various analytes in different fields. As can be seen, the dynamic range, limit of detection, precision, and throughput (—80-100 samples/ h) are all quite good. All determinations are based on the use of the TCPO/ hydrogen peroxide system by exception, that for p-carboline alkaloids uses TCPO and DNPO. A comparison of the analytical figures of merit for these alkaloids reveals that DNPO results in better sensitivity and lower detection limits. However, it also leads to poorer precision as a result of its extremely fast reactions with the analytes. Finally, psychotropic indole derivatives with a chemical structure derived from tryptamines have also been determined, at very low concentrations, by CAR-CLS albeit following derivatization with dansyl chloride. 

Enhancements in the sensitivity with which amino acids containing a primary amine group can be determined have been achieved by derivatization. Chen and Sato [37] reported derivatization with divinyl-sulfone-reduced limits of detection by several orders of magnitude, while Lee and Nieman [38] reported derivatization with dansyl-chloride-reduced limits of detection by a factor of three. 

Dansyl chloride METQL H202-dansyl chloride pH 7.0... 

Figure 14 Representative fluorescence derivatization reagents used for PO-CL detection. DNS-CI, dansyl chloride DBD-F, 4-(A,A-dimethylaminosulphonyl-7-fluoro-2,l, 3-benzoxadiazole NDA, naphthalene-2,3-dicarboxaldehyde DNS-H, dansyl hydrazine DBD-H, 4-(iV,iV-dimethylaminosulphonyl-7-hydrazino-2,l,3-benzoxadiazole).

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