Abstract
The variation of contents of free amino acids in unhulled wheat and barley, during the ripening period of their plants, was investigated in detail by the combined use of ion exchange chromatography and paper partition chromatography.
Total free amino acids adsorbed on a column of Amberlite-IR-120 (H)(2×25cm) were thoroughly eluted with aqueous ammonia (5%), and the combined eluate was taken to dryness in vacuo, after which the residue was dissolved in10ml of distilled water.
An aliquot of 0.001ml of this solution was chromuographed on a sheet of paper by the two-dimensional, ascending development method, using 0.1% acetonic solution of ninhydrin as a chromogenic spray reagent.
Each spot of chromatograms of amino acids was cut off, and to this was added 2ml of 10% of aqueous solution of ninhydrin and1ml of 10% aqueous solution of pyridine, and the solution was boiled on a water-bath for 25 min.
Each of the colored eluates was subjected to spectrophotometric determination at 570mμ(in the case of proline and hydroxyproline at 440 mp).
The results were tabulated in Tables 2 and 3.
