Zhen Qianna, Huang Xiaolan, Zhang Xiaoqing, Ban Yanna, Ding Min
Department of Laboratory Medicine, Chongqing Medical University, Key Laboratory of Clinical Laboratory.
Se Pu. 2011 May;29(5):435-8. doi: 10.3724/sp.j.1123.2011.00435.
A high performance liquid chromatographic (HPLC) method with programmed wavelength ultraviolet (UV) detection was established to simultaneously determine tryptophan (Trp) and its key metabolites kynurenine (Kyn) and 5-hydroxytryptamine (5-HT). A BDS-Hypersil-C8 column (150 mm x 4.6 mm, 5 microm) was used for the analysis at 25 degrees C. The separation was carried out with the mobile phase consisting of 10 mmol/L sodium acetate-acetic acid (pH 4. 5) and acetonitrile (94: 6, v/v) using theophylline as internal standard (IS) at a flow rate of 0.6 mL/min. The eluates were monitored by the programmed wavelength UV detection at 360 nm for Kyn and IS, 220 nm for 5-HT and 302 nm for Trp. The mean recoveries were in the range of 87% to 113%. The linearities were from 3.97 to 400 micromol/L for Trp, 0.421 to 20.2 micromol/L for Kyn and 4.36 to 980.5 nmol/L for 5-HT. The detection limits were 0.134 micromol/L for Trp, 0.016 micromol/L for Kyn and 2.03 nmol/L for 5-HT. Fifteen plasma samples of patients with depression and fifteen plasma samples of healthy controls were tested, and the results demonstrated that the metabolism of Trp in the plasma from the depression group was significantly different from that of the control group.
建立了一种采用程序波长紫外（UV）检测的高效液相色谱（HPLC）方法，用于同时测定色氨酸（Trp）及其关键代谢产物犬尿氨酸（Kyn）和5-羟色胺（5-HT）。使用BDS-Hypersil-C8柱（150 mm×4.6 mm，5μm）在25℃下进行分析。分离采用流动相，流动相由10 mmol/L醋酸钠-醋酸（pH 4.5）和乙腈（94:6，v/v）组成，以茶碱作为内标（IS），流速为0.6 mL/min。洗脱液通过程序波长紫外检测进行监测，Kyn和IS在360 nm处检测，5-HT在220 nm处检测，Trp在302 nm处检测。平均回收率在87%至113%范围内。Trp的线性范围为3.97至400μmol/L，Kyn为0.421至20.2μmol/L，5-HT为4.36至980.5 nmol/L。Trp的检测限为0.134μmol/L，Kyn为0.016μmol/L，5-HT为2.03 nmol/L。对15例抑郁症患者的血浆样本和15例健康对照的血浆样本进行了检测，结果表明抑郁症组血浆中Trp的代谢与对照组有显著差异。
