Zhen Qianna, Xu Biao, Ma Li, Tian Gang, Tang Xiufang, Ding Min
Clin Biochem. 2011 Feb;44(2-3):226-30. doi: 10.1016/j.clinbiochem.2010.10.011. Epub 2010 Nov 17.
To develop a reliable HPLC method for the simultaneous determination of plasma tryptophan, kynurenine and 5-hydroxytryptamine to analyze tryptophan metabolism.
Separation was carried out on a C8 column with the mobile phase composed of acetate buffer (pH 4.5) and acetonitrile using theophylline as internal standard. The eluates were monitored by ultraviolet detection with programmed wavelength.
Analysis was achieved in less than 8.0min. The limits of quantification were 3.97μmol/L, 4.36nmol/L and 0.421μmol/L for tryptophan, 5-hydroxytryptamine and kynurenine, respectively. Reproducibility and recovery were satisfactory. Twenty healthy adults and 20 uremic patients undergoing hemodialysis were analyzed using the present method. Tryptophan metabolism was found to be disturbed in uremic patients and was improved obviously after hemodialysis.
The developed HPLC method is simple, reliable and suitable for monitoring tryptophan metabolism in uremic patients undergoing hemodialysis.
建立一种可靠的高效液相色谱法，用于同时测定血浆色氨酸、犬尿氨酸和5-羟色胺，以分析色氨酸代谢。
在C8柱上进行分离，流动相由醋酸盐缓冲液（pH 4.5）和乙腈组成，以茶碱作为内标。洗脱液通过程序波长紫外检测进行监测。
分析在不到8.0分钟内完成。色氨酸、5-羟色胺和犬尿氨酸的定量限分别为3.97μmol/L、4.36nmol/L和0.421μmol/L。重现性和回收率令人满意。使用本方法对20名健康成年人和20名接受血液透析的尿毒症患者进行了分析。发现尿毒症患者的色氨酸代谢紊乱，血液透析后明显改善。
所建立的高效液相色谱法简单、可靠，适用于监测接受血液透析的尿毒症患者的色氨酸代谢。
