Ninhydrin react with amino

Amino acids are colorless, so a detection technique is necessary in order to visuafize the location of the various spots. The most common method involves treating the filter paper or gel with a solution containing ninhydrin followed by heating in an oven. Ninhydrin reacts with amino acids to produce a purple product. 

The post-column ninhydrin reaction proceeds in a reaction coil at elevated temperatures (ca. I30-I35°C). Ninhydrin reacts with primary amino acids to give a chromophore with a wavelength of maximum absorption of 570nm. 

Fluorimetric methods for the determination of amino acids are generally more sensitive than colorimetric methods. Fluorescamine (4-phenyl-spiro[furan-2(3H),l -phthalan]-3,3 -dione) and o-phthaldialdehyde (OPA) substances are used for protein analysis. Fluorescamine reacts with amino groups to form fluorophores that excite at 390 nm and emit at 475 nm (Weigele et al., 1972). Applications of fluorescamine include monitoring the hydrolysis of K-casein (Beeby, 1980 Pearce, 1979) and quantification of proteins, peptides, amino acids in extracts (Creamer et al., 1985). OPA produces fluorescence on reaction with 2-mercaptoethanol and primary amines, with strong absorption at 340 nm. Lemieux et al. (1990) claimed that this method was more accurate, convenient, and simple for estimating free amino acids than the TNBS, ninhydrin, or fluorescamine methods. 

Ninhydrin is a common reagent for visualizing spots or bands of amino acids that have been separated by chromatography or electrophoresis. When ninhydrin reacts with an amino acid, one of the products is a deep violet, resonance-stabilized anion called Ruhemann s purple. Ninhydrin produces this same purple dye regardless of the structure of the original amino acid. The side chain of the amino acid is lost as an aldehyde. 

In the amino acid analyzer, the components of the hydrolysate are dissolved in an aqueous buffer solution and separated by passing them down an ion-exchange column. The solution emerging from the column is mixed with ninhydrin, which reacts with amino acids to give the purple ninhydrin color. The absorption of light is recorded and printed out as a function of time. 

Amino Group Content. The relative concentration of (a +e)-amino groups was determined colorimetrically by reaction with ninhydrin (20). In this method, the ninhydrin reacts with the a- and e-amino groups in silk to form a blue-colored compound having a maximum absorption at 570 nm. 

The second method involves staining larval entry holes in pulse seeds. In the third method, ninhydrin (triketohydrindene hydrate) has been used to react with the body fluids of insects developing inside grains. A purple spot develops when ninhydrin reacts with the free amino acids (and keto acids) present in the body fluids of insects (Dennis and Decker, 1962). Ninhydrin at a 0.3% level in acetone is impregnated into filter paper. When infested grains are crushed between the folds of ninhydrin-treated paper, the body fluids from the crushed insect bodies react with ninhydrin in the paper. Purple spots develop in the paper in 20-30 minutes at room temperature (20-25 °C), the number of spots indicating the number of insects present. 

Ninhydrin reacts with an amino acid to form a purple-colored compound. Propose a mechanism to account for the formation of the colored compound. 

A principal question, the monitoring of the effluent from the column remains to be discussed here. Small amounts of the individual amino acids emerging in the sequence of their elution have to be revealed and quantitated. In earlier procedures these amounts were in the range of 10 to 100 nanomoles but with improvements in the methodology much smaller samples can now be applied. The classical color reagent for the detection of nanomole quantities is ninhydrin, which reacts with amino acids in a transamination-decarboxylation reaction to yield Ruhemann s purple ... 

Udenfriend and co-workers showed that phenylacelal-dehyde, which is formed by oxidative decarboxylation of phenylalanine by ninhydrin, reacts with excess of ninhydrin and with a primary amino group to give a fluorescent product [228]. The elucidation of this mechanism and of the structure of the fluorescent reaction product [229] led to the synthesis of fluorescamine [230]. 

The simplest method for the determination of amino acids is reaction with ninhydrin. Ninhydrin reacts with both primary and secondary amino acids to produce Ruhemann s purple, which can be detected by ultraviolet (UV)-visible spectroscopy. The reaction requires heat, and a reducing agent is generally added to stabilize the color formation. Primary amines are detected with the greatest sensitivity at 570 nm, while the absorption maximum for secondary amines is 440 nm. If both primary and secondary amines are to be determined, a common absorption wavelength of 500 nm is employed however, this leads to decreased sensitivity. Under optimal... 

A dye commonly used to detect amino acids is ninhydrin (1,2,3-indanetrione monohydrate). Ninhydrin reacts with a-amino acids to produce an aldehyde, carbon dioxide, and a purple-colored anion ... 

Amino add raagants reagents for the colorimetric identification and quantitation of amino acids. One of the most important is ninhydrin (2,2-dihy-droxy-lH-indene-13(2H)-dione), which reacts with amino acids to form a blue-violet dye called Ruhe-mann s purple (absorbance maximum 570 nm). With the imino acid, proline, ninhydrin forms a yellow product, absorbance maximum 440 nm. 

Even today, the postcolumn derivatization with ninhydrin introduced by Spademan, Stein, and Moore [29] represents the most common detection method for quantitative amino acid analysis. As a strong oxidant, ninhydrin reacts with the a-amino groups of eluting amino acids at temperatures around 130 °C, according to Eq. (5.3), releasing carbon dioxide. 

T. Wieland I would say that the reaction of glutathione with ninhydrin is a reducing one and in this reaction glutathione is oxidized to its disulfide. Ninhydrin reacts with the free amino group of glutathione and it may be possible that in this way are formed many products in the same reaction of glutathione with ninhydrin at least. . . ... 

Ninhydrin (Section 23.3) The hydrate of indan-l,2,3-trione, a molecule that reacts with amino acids to give the purple dye used in quantitative analysis of amino acids. 

Ninhydrin is a compound that reacts with amino acids and proteins to produce a dark-colored complex. It is used by forensic chemists and detectives to see fingerprints that might otherwise be invisible. Ninhydrin s composition is 60.68% carbon, 3.40% hydrogen, and 35.92% oxygen. What is the empirical formula for ninhydrin ... 

The amino acid analyzer (Section 26-5) detects the elution of any amino acid by the presence of an indicator that turns a deep violet color. This indicator is ninhydrin A, which reacts with amino acids to the purple compound B ( Ruhemaim s purple ), generating as by-products an aldehyde and CO2. 

The amino acids probably react with the fluorescein isothiocyanate to yield fluorescein thiourea derivatives. These are hydrolyzed at elevated temperature in alkaline medium so that the amino groups that are produced can then react with ninhydrin. 

Detection powders and fingerprint development kits commonly contain cream- or yellow-colored ninhydrin crystals or a solution of dissolved ninhydrin. Ninhydrin (also known as 1,2,3-indantrione, monohydrate 2,2-dihydroxy-1,3-indandione triketohydrindene, monohydrate and triketohydrinden hydrate) has the structure presented in Fig. 13.3.1. Ninhydrin will react with a free a-amino group, -NH2. This group is contained in all amino acids, and analysis with ninhydrin is often performed to verify the presence of amino acids. When a-amino acids (i.e., amino acids with the structure NH2-CHR-COOH) react with ninhydrin, a characteris-... 

D2. Dent, C. E., A study of the behaviour of some sixty amino acids and other ninhydrin reacting substances on phenol-collidine filter paper chromatograms with notes as to the occurrence of some of them in biological fluids. Biochem. J. 43, 169-180 (1948). 

With secondary amino acids, ninhydrin reacts to form a yellow complex with different absorbance characteristics. For this reason, detection occurs at both 570 and 440nm. 

FIGURE 5.5 Schedule for the solid-phase synthesis of somatostatin, a 14-mer, on 10 g of resin reacted with 5 mequiv of the first amino acid, adapted from J. Rivier, J. Am. Chem. Soc. 96 2986, 1974. Min = time of mixing X2 = two times DCC = dicyclohexylcarbodiimide. Step 4 included 5% of (CH2SH)2 to prevent the oxidation of tryptophan. When the ninhydrin test on an aliquot after step 13 was negative, step 1 followed when positive, steps 9-13 were repeated. 

Ninhydrin (triketohydrindene hydrate) reacts with an amino acid when heated under acidic conditions (pH 3-4) to produce ammonia, carbon dioxide and a blue-purple complex. This reaction forms the basis of many widely used methods (Figure 10.11). One mole of carbon dioxide is liberated from each mole of amino acid, exceptions being the dicarboxylic amino acids, which produce two moles of carbon dioxide, and the a-imino acids, proline and hydroxyproline, which do not produce carbon dioxide. Although this formed the basis of a gasometric technique, colorimetric methods are now the most common. 

All primary amines react with fluorescamine under alkaline conditions (pH 9-11) to form a fluorescent product (Figure 10.12) (excitation maximum, 390 nm emission maximum, 475 nm). The fluorescence is unstable in aqueous solution and the reagent must be prepared in acetone. The secondary amines, proline and hydroxyproline, do not react unless they are first converted to primary amines, which can be done using A-chlorosuccinimide. Although the reagent is of interest because of its fast reaction rate with amino acids at room temperature, it does not offer any greater sensitivity than the ninhydrin reaction. 

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