Dong Qian, Jin Wenrui, Shan Jihao
School of Chemistry and Chemical Engineering, Shandong University, Jinan, China.
Electrophoresis. 2002 Feb;23(4):559-64. doi: 10.1002/1522-2683(200202)23:4<559::AID-ELPS559>3.0.CO;2-G.
The precapillary derivatization of 20 amino acids with naphthalene-2,3-dicarboxaldehyde (NDA) and CN(-) was investigated. All these derivatized amino acids could be oxidized on the carbon fiber microdisk bundle electrode except proline. Capillary zone electrophoresis with electrochemical detection was employed for the analysis of 19 amino acids. The optimum conditions of separation and detection were borate, pH 9.48, for the electrolyte, 18 kV for the separation voltage and 1.15 V versus a saturated calomel electrode for the detection potential. Limits of detection of concentration or mass for individual amino acids were between 1.7 x 10(-7) and 1.8 x 10(-6) mol/L or 84 and 893 amol (according to the signal-to-noise ratio of 3) for the injection voltage of 6 kV and injection time of 10 s. The relative standard deviations were between 0.80 and 2.3% for the migration times and 1.4 and 6.4% for the electrophoretic peak currents. From a mixture of 19 amino acids, 10 amino acids (Arg, Lys, Orn, Try, Ser, Ala, Gly, Cys, Glu, Asp) could be well separated. The other 9 amino acids appeared on three electrophoretic peaks. From the samples, in which the nine amino acids do not exist simultaneously, some of them could also be detected. The method was applied to the determination of amino acids in beer by the standard addition method. The recovery for the amino acids in beer was 91-109%.
研究了用萘-2,3-二甲醛(NDA)和氰根离子对20种氨基酸进行毛细管前衍生化。除脯氨酸外,所有这些衍生化氨基酸均可在碳纤维微盘束电极上被氧化。采用毛细管区带电泳-电化学检测法分析19种氨基酸。分离和检测的最佳条件为:电解液为pH 9.48的硼酸盐,分离电压为18 kV,检测电位相对于饱和甘汞电极为1.15 V。对于6 kV的进样电压和10 s的进样时间,各氨基酸的浓度或质量检测限在1.7×10⁻⁷至1.8×10⁻⁶ mol/L或84至893 amol之间(根据信噪比为3)。迁移时间的相对标准偏差在0.80%至2.3%之间,电泳峰电流的相对标准偏差在1.4%至6.4%之间。从19种氨基酸的混合物中,10种氨基酸(精氨酸、赖氨酸、鸟氨酸、色氨酸、丝氨酸、丙氨酸、甘氨酸、半胱氨酸、谷氨酸、天冬氨酸)能够得到很好的分离。另外9种氨基酸出现在三个电泳峰上。对于不存在9种氨基酸同时存在的样品,其中一些氨基酸也能被检测到。该方法应用标准加入法测定啤酒中的氨基酸。啤酒中氨基酸的回收率为91% - 109%。