【摘要】 合成了３种新的荧光标记试剂；吖啶－Ｎ－乙酰氯（ＡＲＣ－Ｃｌ），咔唑－９－乙酰氯（ＣＲＡ－Ｃｌ）和 咔唑－９－丙酰氯（ＣＲＰ－Ｃｌ）。它们的最大发射和激发波长分别为 ４３０ｎｍ（λｅｘ＝４３０ ｎｍ），３６８ ｎｍ λｅｘ＝３３５ｎｍ ）和 ３６５ｎｍ（λｅｘ＝３４０ｎｍ ）。 ３种衍生化试剂与氨基酸形成的衍生物在ｐＨ ６．５的 条件下结合梯度洗脱程序在 Ｃ１８反相柱上对色谱条件进行了优化。每种氨基酸在 ５ ｐｍｏｌ的浓 度下的相对标准偏差小于４％，检测限达ｆｍｏｌ级。更多还原

【Abstract】 A high-performance liquid chromatographic method for the determination of amino acids with fluorescence detection has been developed. The reactions of three fluorescent tagging reagents, acridone-N-acetyl chloride (ARC-CI), carbazole-9-acetyl chloride (CRA-CI) and carbazole-9-propionyl chloride (CRP-CI), with amino acids are reported. Emission maxima for the ARC, CRC and CRP derivatives of amino acids were 430 nm (λex=404nm), 368 nm (λem= 335 nm) and 365 nm (λex = 340nm), respectively. In all cases, the derivatives exhibited strong fluorescence whereas the reagents themselves also exhibited fluorescence. The libelled derivatives are very stable, less than 4% decomposition occurs after heating at 40C for 24 h. fluorescence intensities of amino acid derivatives were higher in neutral and alkline than in acidic solutions. This method, combining a muti-gradient program, offers baseline resolution of the common ARC, CRA and CRP amino acid derivatives from a linear acetonitrile gradient. Separations of amino acid dervatives were carried out on a reversed-phase C18 column. Derivatization and chromatographic conditions were optimized. Amino acid derivatives were eluted with a good reproducibility. The relative standard deviation (n = 6) at an analytical concentration of 5 pmol of each amino acid is < 4%. The detection limit is around fmol level.更多还原