【摘要】 建立以乙酸缓冲系统和甲醇作流动相、电化学和紫外检测器联用的高效液相色谱法，分离和测定了色氨酸经5-羟色胺和犬尿酸原两条主要代谢途径的8种代谢物。使用三氯乙酸作离子对试剂以延长3-羟犬尿酸原的保留时间，分析了流动相pH值和三氯乙酸浓度对各物质分离的影响及检测条件。结果表明，pH值及三氯乙酸浓度对各物质保留时间有明显影响，可作为控制分离的主要因素。此外，对生物样品中各物质分离和检测条件进行了讨论。更多还原

【Abstract】 A reversed phase HPLC method for separation and determination of the major tryptophan (TRP)metabolites in both pyrrolas pathway and TRP hydroxylase pathway, including TRP, kynurenine(KN), 3hydroxykynurenine (3-HKN), kynurenic acid (KA), xanthurenic acid (XA), 5-hydroxytryptop ban (5-HTP ),5-hydroxytr yp tamin (5-HT ) and 5-hydroxyindoleacetic acid (5-HIAA ), has been de veloped by sequential optimization of mobile phase based on acetate buffer and methanol. Trichloroacetic acid(TCA) was used as ionpairing reagent to increase the retention of 3-HKN. The effects of pH and concentrations of TCA on separation were studied. Good separation can be achieved at pH 4. 0-5. 0 of mobile phase in less than 25 min. When TCA was not used, 3-HKN was hard to be detected in biological samples. The maximum retention of 3-HKN was obtained at pH 4. 0 with mobile phase containing 50 mmol/L TCA. Combination of electrochemical (ED) and ultraviolet (UV) detection was used in which ED was responsible for detection of 3-HKN , 5-HTP,5-HT and 5-HIAA and UV for the others. The innuences of potential of ED and wavelength of UV on detection were studied. The optimization of conditions for separation and detection in different biological samples was also discussed.更多还原