Schmidt J, McClain C J
Division of Gastroenterology, Veterans Administration Medical Center, Lexington, KY 40511.
J Chromatogr. 1987 Aug 7;419:1-16.
Twenty-three o-phthalaldehyde-mercaptoethanol derivatives of primary amino acids in serum were separated with Waters C18 5-microns Radial-Pak Resolve columns, using aqueous phosphate-methanol mobile phase with acetate and tetrahydrofuran as modifiers. Resolution is critical in this system for glutamine/histidine, citrulline/glycine/threonine/3-methylhistidine, and tryptophan/methionine derivatives, and is affected by small changes in column properties or mobile phase. However, pre-run adjustments in gradient scheme and/or mobile phase composition can usually be used to obtain chromatograms in which all derivatives can be quantitated. For this purpose, and for general method development, we have written two very fast, interactive programs for the Zenith Z-100 Microsoft BASIC compiler: RTGRAPH, for retention, separation, and resolution plots; and LCSIM for simulations of multisegment binary gradient elution. These programs are shown here to be useful and accurate in most aspects required for developing strategies for this method.
血清中23种伯氨基酸的邻苯二甲醛-巯基乙醇衍生物，采用沃特世C18 5微米径向Pak Resolve柱进行分离，以含醋酸盐和四氢呋喃作为改性剂的磷酸盐-甲醇水溶液作为流动相。在该系统中，谷氨酰胺/组氨酸、瓜氨酸/甘氨酸/苏氨酸/3-甲基组氨酸以及色氨酸/蛋氨酸衍生物的分离度至关重要，且会受到色谱柱性质或流动相的微小变化影响。然而，通常可通过运行前对梯度方案和/或流动相组成进行调整，来获得能够对所有衍生物进行定量的色谱图。为此，以及为了一般方法的开发，我们为Zenith Z - 100微软BASIC编译器编写了两个非常快速的交互式程序：RTGRAPH，用于保留时间、分离和分离度绘图；LCSIM用于多段二元梯度洗脱的模拟。结果表明，这些程序在开发该方法策略所需的大多数方面都很有用且准确。
