Hariharan M, Naga S, VanNoord T
University of Michigan Medical School, Department of Psychiatry, Ann Arbor 48109-0656.
J Chromatogr. 1993 Nov 17;621(1):15-22. doi: 10.1016/0378-4347(93)80071-b.
A reversed-phase liquid chromatographic method for the separation of 26 phenylthiocarbamyl derivatives of amino acids in human plasma in ca. 35 min. is described. The method used a C18 column (150 x 4.6 mm I.D., 3 micron) thermostatted at 41 degrees C, and a simple multistep linear gradient of two solvents. Solvent A was 0.05 M sodium acetate (pH 5.1)-acetonitrile (98:2, v/v), and solvent B was water-acetonitrile (40:60, v/v). A simple and successful approach to the optimization of the conditions for the separation of the 26 amino acid derivatives was realized. In the initial phase of development, the composition of the gradient, its timings, the column temperature, the flow-rate and the mobile phase compositions were optimized. At the end the influence of pH was studied, and this approach led to a clear resolution of the 26 amino acids. The method was validated by accuracy, precision, and recovery studies, by analyzing patient samples, and by comparing the quality control sample results with the classical ion-exchange method.
描述了一种反相液相色谱法，可在约35分钟内分离人血浆中26种氨基酸的苯硫代甲酰胺衍生物。该方法使用一根在41℃恒温的C18柱（内径150×4.6mm，3微米），以及两种溶剂的简单多步线性梯度。溶剂A为0.05M醋酸钠（pH 5.1）-乙腈（98:2，v/v），溶剂B为水-乙腈（40:60，v/v）。实现了一种简单且成功的优化26种氨基酸衍生物分离条件的方法。在方法开发的初始阶段，对梯度组成、其时间、柱温、流速和流动相组成进行了优化。最后研究了pH的影响，这种方法实现了26种氨基酸的清晰分离。通过准确性、精密度和回收率研究、分析患者样本以及将质量控制样本结果与经典离子交换法进行比较，对该方法进行了验证。
