高效液相色谱-荧光检测法在慢性肾功能不全患者血清芳香族氨基酸检测中的临床价值  

Determination of serum aromatic amino acid in patients with CRI by high performance liquid chromatography with fluorescence detection

作　　者：李影[1] 唐爱国[1] 穆萨[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

出　　处：《中华检验医学杂志》2011年第9期769-774,共6页Chinese Journal of Laboratory Medicine

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

摘　　要：目的建立同时测定血清AAA含量的HPLC—FLD法，探讨CRI患者血清AAA含量变化及其临床应用价值。方法血清标本来自于100名健康体检者和80例CRI患者。将CRI患者按2002年美国肾脏基金会（NKF）诊断分期标准进行分期：CKD2期4例、CKD3期12例、CKD4期12例和CKD5期52例；按CRI不同病因分组：慢性肾炎型32例、糖尿病型36例和高血压型12例。血清经高氯酸去蛋白后，离心取上清液测定，外标法定量。采用MegresC18色谱柱，流动相为乙腈：水（体积比为1：9），流速为1．0ml／min，荧光检测器在不同时间段设定特定波长对血清AAA进行测定。对健康对照组和CRI患者组血清中AAA总量、Tyr、Phe和Trp含量及不同分期和不同病因CRI患者血清Tvr、Phe和Trp含量进行比较，同时评价血清AAA总量诊断CRI的敏感度与特异度。结果Tyr、Phe和Trp线性范围分别为0．550～275．000、3．050～1220．000和0．049～49．000μmol／L，最低检测限分别为0．014、0．500和0．005μmoL／L，平均回收率分别为100．9％、101．3％和98．5％，日内精密度为2．32％～3．92％（平均为3．13％），日间精密度为3．18％～4．20％（平均为3．58％）。CRI患者组血清AAA总量、Tyr、Trp含量及Tyr／Phe比值分别为（135．74±12．23）、（52．27±8．25）、（21．49±4．25）μmol／L和[0．87（0．68～1．05）]，低于健康对照组的（174．47±11．57）、（63．53±4．68）、（44．22±3．67）μmol／L和[0．97（0．94—1．00）]，差异均有统计学意义（t=-14．709、4．452、22．100，U=266．000，P均〈0．05）。不同分期CRI患者Tyr、Phe和Trp含量差异无统计学意义；Tyr含量在慢性肾炎组、高血压组和糖尿病组间差异无统计学意义，Phe在慢性肾炎组与高血压组、慢性肾炎组与糖尿病组问差异有统计学意义（U=114．00、395．00，P均〈0．05），Trp在慢性肾炎组与糖尿病组间差异有统�Objective A HPLC-FLD method was developed to determine the levels of serum AAA in CRI patients, and to study the variation of serum AAA in CRI patients and its clinical significances. Methods Serum samples were collected from 100 health controls and 80 CRI patients. According to 2002 National Kidney Foundation （NKF） staging diagnosis, CRI patients included 4 of stage 2, 12 of stage 3, 12 of stage 4, 52 of stage 5. According to pathogenesis, CR! patients were also divided into 3 groups：chronic nephritis group （n =32）, DM group （n =36）, hypertension group （n = 12）. Serums were deproteinized by equal volume of 5 % （v/v） PCA and supernate were analyzed directly. External standard method was used as quantitative method. The analytical column was Megres C18. 10% acetonitrile in water was used as mobile phase. Flow rate was 1.0 ml/min. The wavelengths of fluorescence excitation and emission were changed with specific time. The levels of Tyr, Phe and Trp in CRI groups, different CKD stages and different pathogenesis were compared with healthy control groups to evaluated the sensitivity and specificity of serum AAA for CRI diagnosis. Results The linear ranges of the method were 0. 550 - 275.000, 3.050 - 1220. 000 and O. 049 - 49.000 μmol/L for Tyr, Phe and Trp, respectively. The limit of detection （LOD） was 0. 014 μmol/L for Tyr, 0. 500μmol/L for Phe, and 0. 005 μmol/L for Trp. The average recovery was 100. 9% , 101.3% and 98. 5% for Tyr, Phe and Trp, respectively. Intra-day CV was 3.18% - 4. 20% （mean was 3.13% ） and inter-day CV was 3.18% - 4. 20% （ mean was 3.58% ）. The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in CRI patients were（ 135.74 ± 23.23 ） μmol/L, （52. 27 ± 8.25） μmol/L, （21.49± 4. 25 ） μmol/L and [ 0. 87 （0. 68 - 1.05 ） ] μmol/L. which were lower than that in healthy groups （ t value was - 14. 709, 4. 452, 22. 100, U value was 266. 000, respectively, P 〈 0.05 ）. The concentration of serum AAA, Tyr and Trp and the ratio

关 键 词：肾功能不全 慢性 氨基酸类 芳香 色谱法 高压液相 

分 类 号：R692.5[医药卫生—泌尿科学]