| 摘要 |
PURPOSE:To realize a method for determination of amino acids except alpha-amino acid having the sensitivity and accuracy equiv. to the sensitivity and accuracy of the conventional method for determination by adding an isothiocyanic aryl to a sample soln. and extracting away the product of reaction with the alpha-amino acid. CONSTITUTION:Ethanol, etc., are first added to a vital sample and protein is removed by centrifugal sepn. A basic buffer soln. is added to the supernatant thereof, then the isothiocyanic aryl is added thereto and the liquid is stirred to effect reaction. A proper amt. of water is added to the liquid after the reaction and the extraction is executed with a weakly alkaline and water insoluble org. solvent so that the excess reagent, etc., are migrated to the org. solvent layer. The water layer is then converted to acidity by a mineral acid and is heated to effect reaction. This reaction liquid is concd. and dried under a reduced pressure and is converted to weak alkalinity by a boric acid buffer soln., etc. the liquid is then subjected to an extraction operation by the org. solvent to migrate the thiohydantoin deriv. of the alpha-amino acid to the org. solvent. The liquid is thereafter converted to acidity by hydrochloric acid, etc., and is then extracted by the org. solvent. The org. solvent is removed from this extraction liquid and a proper volume thereof is subjected as the soln. of the eluate of liquid chromatography to high-performance liquid chromatography, by which the determination of the amino acids with high accuracy is permitted.
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