HPLC法测定末梢血中苯丙氨酸与酪氨酸及在苯丙酮尿症中的应用  

Determination of Phenylalanine and Tyrosine in Peripheral Blood by HPLC and Its Application in Phenylketonuria

作　　者：莫喜明[1] 唐爱国[1] 

机构地区：[1]中南大学湘雅第二医院检验科,长沙410011

出　　处：《现代检验医学杂志》2008年第4期20-23,共4页Journal of Modern Laboratory Medicine

基　　金：湖南省科技厅资助项目(04SK3052)

摘　　要：目的建立一种简单、快速的反相高效液相色谱法测定末梢血液中的苯丙氨酸(Phe)与酪氨酸(Tyr)并用于苯丙酮尿症(PKU)的筛查。方法末梢血标本与等量5%(v/v)高氯酸溶液混合处理去除蛋白后离心取上清液20μl直接进样分析。用Hypersil C8色谱柱(6.0mm×300mm,10μm)分离,流动相为乙腈-水(体积比5∶95),流速1.5 ml/min,紫外检测波长210 nm,室温下测定。结果末梢血中Tyr和Phe均分离良好,保留时间分别为5.88 min和8.43 min。102例健康儿童指血Phe浓度67.7±15.4μmol/L,Tyr浓度为62.2±13.9μmol/L,Phe/Tyr比值为1.15±0.27。32例正常新生儿足跟血Phe浓度为66.2±20.5μmol/L,Tyr浓度为59.5±18.8μmol/L,Phe/Tyr比值为1.12±0.24。结论该法灵敏度高,特异性好,方法简便快速,成本低廉,适合于末梢血中Phe和Tyr的同时测定及PKU的筛查与监控。Objective To establish a simple and rapid method for the determination of phenylalanine and tyrosine in peripheral blood by reverse phase high-performance liquid chromatography （RP-HPLC） ,and explore the feasibility of the method for neonatal screening of phenylketonuria（PKU）. Methods Supernatant fluid of peripheral blood precipitated with perchloric acid was isocratically Muted using a base-deactivated C8 column with 5% （v/v） acetonitrile in water as the mobil phase. Untraviolet detector worked at 210 nm. Results The retention time of tyrosine （Tyr） and phenylalanine （Phe） were 5.88 min and 8.43 min respectively. Phe and Tyr in finger blood of 102 health enfant were （67.7±15.4） μmol/L and （62. 2±13.9） μmol/L respectively. The ratio of Phe/Tyr was 1.15±0.27. Phe and Tyr in heel blood of 32 health neonatal were （66.2±20.5）μmol/L and （59.5±18.8） μmol/L respectively. The ratio of Phe/Tyr was 1.12±0.24. The levels of phenylalanine and tyrosine in whole-blood which anticogulated by EDTA-K2 were consistant with those of anticogulating by heparin（P〉0.05）. The levels of phenylalanine and tyrosine in peripheral blood were significantly lower than those in plasma, but showed a good correlation. Phenylalanlne and tyrosine in whole-blood which storaged in frost were more stability in one week. Conclusion This method is sensitivity,accurate ,rapid and simple in measuring phenylalanine and tyrosine in finger blood.

关 键 词：高效液相色谱 苯丙氨酸 酪氨酸 苯丙酮尿症 新生儿筛查 

分 类 号：R722.11[医药卫生—儿科] Q503[医药卫生—临床医学]