摘要
建立了一种同时测定人体胸腔积液中酪氨酸(Tyr)、苯丙氨酸(Phe)和色氨酸(Trp)的液相色谱分析方法,并将其应用于胸腔积液病因鉴别诊断。优化的液相色谱条件如下:VertexTMAQ-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-10 mmol/L醋酸铵缓冲溶液(醋酸调至pH 5.6)为流动相进行梯度洗脱,流速1.0 mL/min,检测波长为210和278 nm。结果表明,3种氨基酸在各自相应的线性范围内线性相关系数均大于0.999;信噪比为3时,检出限(LOD)分别为0.50,0.17和0.20μmol/L。胸腔积液样品经甲醇去除蛋白质后取上清液直接测定,3种氨基酸平均回收率在80.7%～101.1%之间,相对标准偏差为2.5%～8.7%。利用本方法对72例良恶性胸腔积液中的3种氨基酸进行分析,发现Tyr,Phe和Trp含量在不同疾病的样品中存在统计学差异,对胸腔积液病因鉴别诊断有一定的临床价值。
A HPLC method was developed for the simultaneous determination of tyrosine,phenylanine and tryptophan in pleural effusion and applied to the differential diagnosis of pleural effusion causes.Pleural effusion was employed for the experimental conditions optimizing and the results were as follows： using VertexTM AQ C18（250 mm×4.6 mm,5 μm） as separation column,acetonitrile-10 mmol/L ammonium acetate（adjust pH to 5.6 with acetic acid） as the mobile phase with a gradient elution at the flow rate of 1.0 mL/min and detection wavelength at 210 nm and 278 nm.The result showed that good linear relationship was achieved between the chromatographic peak area and the concentration of each component.The linear correlation coefficient of these three substances was above 0.999.The relative standard deviation of seven parallel injections was less than 3.2%.The limits of detection（LOD） of these three substances were 0.50,0.17 and 0.20 μmol/L,respectively.Pleural effusion samples were precipitated with methanol,then centrifuged to remove protein residue and finally analyzed by HPLC.The recoveries were ranged from 80.7% to 101.1%,the relative standard deviation of five parallel injection was less than 8.7%.The method was applied to the analysis of 72 patients with lung cancer（20 patients） and benign lung diseases（52 patients）.The results indicated that the difference of these three amino acids in different patients was statistically significant and had certain clinical value in differential diagnosis of pleural effusion cause.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2013年第9期1385-1390,共6页
Chinese Journal of Analytical Chemistry
基金
浙江省自然科学基金(Nos.M203030,Y4110536)
浙江省公益性技术应用研究(分析测试)项目(Nos.2011C37040,2011C37009)
浙江省重点科技创新团队(No.2011R09002-11)
浙江省重中之重学科开放基金(No.200908)资助
关键词
高效液相色谱
氨基酸
胸腔积液
鉴别诊断
High performance liquid chromatography
Amino acids
Pleural effusion
Differential diagnosis
