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Analisis Asam Amino dengan Kromatografi Cairan Kinerja Tinggi Secara Derivatisasi Prakolom dan Pascakolom

Wayan Rediatning S. & Nanny Kartini H.

 


Abstract.

Through an experiment process, an optimum condition of amino acid analysis using High Performance Liquid Chromatography (HPLC) with post- and pre-column derivatization methods has been found. The optimum condition of the analysis for the post-column deriyatization was obtained by using resin cation exchanger (R-SO3Na) as the column, citric acid buffer of the pH 3.0 and boric acid buffer of the pH 9.7 as the mobile phase and O-phtalaldehyde (OPA) /2-mercaptoethanoal as the derivator, In the pre-column derivatization method, Resolve 5μ spherical C18 was used as the column, solution A (methanol : THF ; H2O = 2 : 2 : 96) which contains 0.05 M Na2HPO4 and 0.05M NaOAc) and solutionB (methanol:H2O= 65 : 35) as the mobile phase, and OPA/ethantiol as the derivator. Each of both HPLC methods can separate more than 17 kinds of amino acids. The result shows that the pre-column derivatization method was more sensitive and took a shorter analysing time than the post-column derivatization method; however, secondary amino acid could not be determined by the first method.



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