Application of iodine-azide reaction for detection of amino acids in thin-layer chromatography |
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Authors: | Kaźmierczak Dorota Ciesielski Witold Zakrzewski Robert Zuber Monika |
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Affiliation: | Department of Instrumental Analysis, University of Lód?, Pomorska 163, 90-236 Lód?, Poland. |
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Abstract: | The iodine-azide reaction was employed to TLC detection of sulphur-containing derivatives of protein and some non-protein amino acids. The derivatization reaction with phenyl isothiocyanate (PITC) took place directly on the plate before the developing step. Subsequently, the plates were sprayed with a mixture of sodium azide and starch solution in NP-TLC and in the case of RP-TLC sodium azide solution with starch incorporated into mobile phase and then exposed to iodine vapour. The spots became visible as white spots on violet-grey background. The obtained detection limits of PTC-derivatives have been compared with other visualizing techniques commonly used in TLC practice (UV254 and iodine vapour). The iodine-azide system has been proved to be the most favourable and enabled to detect quantities per spot in the range of 1-60 pmol (HPTLC) and 3-100 pmol (TLC). |
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