Sensitive spot test of proline on clay thin layer

GOSHIMA Fumiaki, MIO Sumiko

doi:10.2116/bunsekikagaku.25.805

A sensitive spot test of proline on a sheet of thin layer of Japanese acid clay was investigated. The procedure is as follows: A sample solution (5 μl) of amino acids was placed on a sheet of thin layer of thin layer of, Japanese acid clay.<BR>After drying, a ninhydrin reagent (0.5% ninhydrin in 95% ethanol) was sprayed on that plate, which then was heated at ( 100110) °C for 5 minutes. A yellow spot of proline appeared. After cooling, the plate was sprayed with 2% Na₂SO₃ and then reheated at the same temperature for 10 minutes. The yellow spot turned reddish purple similar to that produced by hydroxyproline. This method was more sensitive than an ordinary method and the limit of detection of the colored spot is 0.05 μg of proline. Then the colored spot were treated with a drop of 1 N NaOH. A reddish purple spot of proline remained, while the colored spot of other amino acids disappeared. However, this specific colored spot of proline also faded gradually.<BR>A purple spot remained on the plate faintly when more than 10 μg of hydroxyproline was present in the sample solution. In this way, 2.5 μg of proline in 5 μl of a mixture could be detected by the reddish purple color in the presence of 4 times amounts of hydroxyproline. Therefore, this reaction provided a method of a sensitive spot test of micro amounts of proline in amino acids mixture.

Sensitive spot test of proline on clay thin layer

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