Separation of Amino acid Biomarkers from Plasma by Mixed Micelle combined with Polyvinylpyrrolidone-Filled Capillary Electrophoresis

• Main Authors: Yen-Chu Chen, 陳彥竹
• Other Authors: Po-Ling Chang
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/43419464548407413591

碩士 === 東海大學 === 化學系 === 102 === Physiological amino acids (AAs) are one of the important indexes for monitoring various diseases such as cancers. This study proposed a polymer-based separation method for the determination of AAs by capillary electrophoresis with light-emitted diode induced fluorescence. The derivatization of AAs withnaphthalene-2,3- dicarboxaldehydeformed an amino acid-cyano [f] benzoisoindole (AA-CBIs) derivative that served as fluorophores for fluorescence detection. The separation of AA-CBIs derivatives was successfully achieved using a solution of polyvinylpyrrolidone (5 % w/v, Mavg 1,300,000 Da) containing 20 % isopropanol and 10 mM HEPES (pH 7.5) buffer solution. To shorten analysis time, sodium dodecyl sulfate from inlet buffer (cathode) was used for to accelerate the velocity of AA-CBIs derivatives as a result of micelle sweeping. Except for proline, all of the AA-CBIs (including high isoelectric point of lysine and arginine) were well separated under the optimized separation condition. Therefore, these results demonstrated that the distinct advantages of the proposed method is simple, highly efficient, and cost-effective, which provides great potential for the diagnosis of several important diseases include carcinomas.