Application of the Iodine–azide Procedure for the Detection of Glycine, Alanine, and Aspartic Acid in Planar Chromatography

Zakrzewski, Robert; Ciesielski, Witold; Kaźmierczak, Dorota

doi:10.1081/jlc-120005707

Cited by 8 publications

(3 citation statements)

References 10 publications

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“…The new proposed derivatization procedure shortens the analysis time and simplifies the derivatization procedure compared with the Edman procedure [3,6].…”

Section: Discussionmentioning

confidence: 99%

Detection of proline, arginine, and lysine using iodine‐azide reaction in TLC and HPTLC

Zakrzewski¹,

Ciesielski

Kaźmierczak

2003

J of Separation Science

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The application of a modified iodine-azide procedure for the detection of proline, arginine, and lysine is described. Phenyl isothiocyanate was used to transform amino acids into phenyl thiocarbamyl derivatives (derivatization in situ). The developed plates were sprayed with a mixture of sodium azide and starch solution, adjusted to pH 5.5, and exposed to iodine vapour. Due to the catalytic effect of the C=S bond, the spots appeared white on a violet-grey background and were stable for 20 minutes. The detection limits were found to lie in the pmole range. The iodine-azide test is compared with other procedures (iodine, UV, ninhydrin).

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“…The new proposed derivatization procedure shortens the analysis time and simplifies the derivatization procedure compared with the Edman procedure [3,6].…”

Section: Discussionmentioning

confidence: 99%

Detection of proline, arginine, and lysine using iodine‐azide reaction in TLC and HPTLC

Zakrzewski¹,

Ciesielski

Kaźmierczak

2003

J of Separation Science

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“…• mobile phase 1: isopropyl alcohol -water (10:90) [7,13]; • mobile phase 2: isopropyl alcohol -water (70:30) [7,13]; • mobile phase 3: ethanol (96 %) -water (70:30) [7,13,25]. To select the most optimal mobile phase, the mixture of glutamic acid and glycine, Reference solution (RS-3), was used.…”

Section: The Tlc-methods Developmentmentioning

confidence: 99%

The Selection of “Green” Conditions for Components Identification in a Combined Medicine by TLC/HPTLC Methods

Rudakova

Gubar

Smielova

et al. 2022

J. Org. Pharm. Chem.

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Aim. To select “green conditions” for identifying components in a combined medicine for the treatment of alcohol intoxication.Materials and methods. Thin-layer chromatography and high performance thin-layer chromatography methods were used. An analytical GREEnness calculator was applied to assess the environmental friendliness of the analytical procedure.Results and discussion. The choice of mobile and stationary phases that comply with the principles of “green chemistry” and can be used to detect glutamic acid and glycine in the composition of a combined medicine has been substantiated. It has been determined that by the indicators Rf, Rs, ΔRf, α, N, H the most effective for the division is the ethanol (96 per cent) – water (70:30) mobile phase (the length of the solvent front is 10 cm, the application volume is 5 μL), which allows, in addition to amino acids, to determine another prescription component – ascorbic acid. The conditions for identification of substances by the high-performance thin-layer chromatography method (the length of the solvent front is 7 cm, the application volume is 2 μL) have been selected. It has been found that to detect chromatographic zones, it is optimal to use ninhydrin solution R1 with further heating of the plate at a temperature of 100-105°С for 5 min. The specificity of determination of glutamic acid, glycine and ascorbic acid in comparison with solutions of standard substances has been proven. While studying the robustness of the method the influence of chromatographic conditions on the final result (the impact of the stationary phase type, chamber saturation, application volume, distance from the “start line” to the “finish line”, the effect of the detection solution, stability of solutions for application) has been researched. The precision of the method on one and three plates of the same type has been studied; the intermediate precision has been researched. The calculated assessment of greenness of the analytical procedure is 0.66.Conclusions. As a result of the studies conducted, “green conditions” for identifying amino acids (glutamic acid, glycine), as well as ascorbic acid in a combined medicine by thin-layer chromatography and high-performance thin-layer chromatography methods have been selected. The validation characteristics of the method (specificity, robustness and precision) have been studied.

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“…In the case of proline, reaction with iodine‐azide is proposed. This reagent is effective for detection of different amino acids including proline and hydroxyproline (Kaźmierczak et al ., ; Zakrzewski et al ., ). Another reagent which is very useful because it produces a stable color with many amino acids is 2,3‐dichloro‐1,4‐naphthoquinone, developed by Samanta and Laskar ().…”

Section: Thin‐layer Chromatographymentioning

confidence: 99%

Application of TLC, HPLC and GC methods to the study of amino acid and peptide enantiomers: a review

Dołowy

Pyka

2013

Biomedical Chromatography

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Amino acids are very important organic compounds in nature. The biological activity of amino acids depends mainly on their stereoisomeric configuration (d- or l-). Thus, the stereochemical analysis of amino acids and peptides is an important aspect of their characterization. Owing to the increasing role of amino acid configuration in biomedical and pharmaceutical studies, numerous analytical methods have been described in the literature. Among a wide range of analytical techniques available for the steroselective separation of different amino acids, which were obtained from plants or biological samples, chromatographic methods such as thin-layer chromatography, high-performance chromatography and also gas chromatography are very useful. This review presents such systems developed for direct stereoisomeric separation and quantitative determination of amino acid and peptide enantiomers with emphasis on selected literature published during two last decades. Almost all aspects, including sample preparation prior chromatographic analysis, stationary phase, solvent system and detection system, are discussed. New possibilities in chiral amino acid analysis have been opened up by the application of mass spectrometry and infrared detection in thin-layer chromatography and mass spectrometry-selected ion monitoring detection system in gas chromatography. A modern trend in high-performance chromatography is two-dimensional chromatography. These innovations have led to decreased development time and increased amino acid resolution and detectability.

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Application of the Iodine–azide Procedure for the Detection of Glycine, Alanine, and Aspartic Acid in Planar Chromatography

Cited by 8 publications

References 10 publications

Detection of proline, arginine, and lysine using iodine‐azide reaction in TLC and HPTLC

Detection of proline, arginine, and lysine using iodine‐azide reaction in TLC and HPTLC

The Selection of “Green” Conditions for Components Identification in a Combined Medicine by TLC/HPTLC Methods

Application of TLC, HPLC and GC methods to the study of amino acid and peptide enantiomers: a review

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