HPLC-FD法测定重性抑郁障碍患者血清酪氨酸、色氨酸和5-羟色胺  

Simultaneous determination of tyrosine,tryptophan and 5-hydroxytryptamine in serum of patients with major depressive disorder by HPLC-FD

作　　者：穆萨[1] 李影[1] 唐爱国[1] 肖乐东[2] 

机构地区：[1]中南大学湘雅二医院检验科,长沙410011 [2]邵阳市第一人民医院检验科,湖南邵阳422001

出　　处：《临床检验杂志》2011年第3期161-164,共4页Chinese Journal of Clinical Laboratory Science

基　　金：湖南省科技计划项目课题(05SK3026;2010SK3205);湖南省医药卫生科研计划课题(C2005019);中南大学研究生教育创新工程项目(2010ssxt070)

摘　　要：目的建立一种高效液相色谱-荧光法(HPLC-FD)同时测定重性抑郁障碍(MDD)患者血清中酪氨酸(Tyr)、色氨酸(Trp)和5-羟色胺(5-HT)的含量,并探讨其应用价值。方法血清标本去蛋白质后,取上清液直接测定。色谱柱采用AtlantisC18(4.6 mm×150 mm,5μm);流动相为0.1 mol/L KH2PO4∶甲醇(85∶15);流速为1.0 mL/min;荧光检测器激发波长和发射波长0～4 min分别为228 nm和306 nm,4～7.5 min为278 nm和338 nm,7.5 min后为285 nm和353 nm。用建立的方法检测MDD患者和健康人对照组血清Tyr、Trp和5-HT含量。结果 Tyr、5-HT和Trp的保留时间约为3、5和9 min,3者分离良好(R>1.5);线性范围分别为0.55～275.00、0.094～47.03和0.049～49.00μmol/L;最低检测限分别为0.014、0.024和0.005μmol/L;平均回收率分别为100.6%、99.3%和97.0%;日内、日间精密度均<5%。苯丙氨酸(Phe)、犬尿氨酸(Kyn)、犬尿喹啉酸(Kyna)等物质无干扰。MDD患者血清Tyr、Trp和5-HT含量均低于健康人对照组,5-HT/Trp比值明显降低,差异有统计学意义(P均<0.01)。结论建立的HPLC-FD方法简便、快速、特异性好,适用于临床检验与研究。测定血清中Tyr、Trp和5-HT含量对MDD的辅助诊断有一定价值。Objective A method was developed for simultaneous determination of tyrosine（Tyr）,tryptophan（Trp） and 5-hydroxytryptamine（5-HT） in serum of the patients with major depressive disorder（MDD） by high-performance liquid chromatography with fluorescence detection（HPLC-FD）,and its value in clinical application was explored.Methods The analytical column was Atlantis C18（4.6 mm×150 mm,5 μm）.The mobile phase was 0.1 mol/L KH2PO4 and methanol（85∶15）.The flow rate was 1.0 mL/min.The wavelengths for fluorescent excitation and emission were operated at 228 nm and 306 nm in 0-4.5 min,278 nm and 338 nm in 4.5-8 min,285 nm and 353 nm after 8 min,respectively.The concentrations of Tyr,Trp and 5-HT of MDD patients and healthy subjects group were determined by HPLC-FD.Results The retention time of Tyr,5-HT and Trp was about 3,5 and 9 min,respectively.The results of separation were satisfied（R1.5）.The linear ranges were 0.55-275,0.094-47.03 and 0.049-49 μmol/L,and the lowest detection limits were 0.014,0.024 and 0.005 μmol/L respectively.The average recovery rates were 100.6%,99.3% and 97.0% respectively.The intra-day and inter-day precision were less than 5%.No interference from Phe,Kyn and Kyna was found.Compared with the healthy controls,the concentrations of serum Tyr,Trp,5-HT and 5-HT/Trp ratio in MDD patients were significantly decreased（P0.01）.Conclusions The method developed in this study is simple,rapid,specific and suitable for clinical determination and scientific research.Simultaneous determination of Tyr,Trp and 5-HT concentrations in serum of MDD patients may have potential clinical value for diagnosis of MDD.

关 键 词：色谱法 高压液相 荧光检测 酪氨酸 色氨酸 5-羟色胺 重性抑郁障碍 

分 类 号：R446.1[医药卫生—诊断学]