摘要
目的建立同时检测血清犬尿喹啉酸(kynurenic acid,KYNA)和色氨酸(tryptophan,Trp)的高效液相色谱一荧光法。方法采用高效液相色谱在线衍生技术,通过对最佳检测波长、流速、流动相中醋酸锌浓度和乙腈比例等因素的探讨,得出测定 Trp 和 KYNA 的最优实验方案,并对其进行方法学评价;测定50名正常人血清 KYNA 和 Trp 含量。结果 KYNA 保留时间约为8.1 min,线性范围为1.05～2093 nmol/L,最低检出浓度为0.05 nmol/L,平均回收率为101.19%;Trp 保留时间约为11.3 min,线性范围为0.49～196μmol/L,最低检出浓度为0.001μmol/L,平均回收率为104.43%,二者日内、日间测定的变异系数均小于5%,苯丙氨酸、酪氨酸、5-羟色胺和犬尿氨酸等物质对该法均无干扰。健康成人血清 KYNA 和 Trp 含量分别为(24.25±9.11)nmol/L和(49.05±11.67)μmol/L。结论建立的方法简便、快速、稳定、可行,适用于临床和科研工作。
Objective To establish a method for simultaneous determination of kynurenic acid （KYNA） and tryptophan （Trp） in serum by high performance liquid chromatography. Methods Chromatographic and detection conditions were optimized and performances of the method were evaluated. Serum KYNA and Trp levels in 50 healthy people were measured with the HPLC method. Results The retention times of KYNA and Trp were 8. 1 min and 11.3 min. The detection limits for the two analytes were 0. 05 μmol/L and 0. 001μmol/L and the linearity ranges from 1.05 nmol/L to 2 093 nmol/L and from 0. 49 μmol/L to 196 μmol/L. The average recovery was 101.19% and the within-run and between-run coefficients of variations were 3.20% and 4.27% for the KYNA analysis, and 104.43%, 3.31% and 4. 14% for Trio. Kynurenin and other similar substances did not interfere with the analyses. The serum KYNA and TRP levels in 50 healthy people were （ 24. 25± 9. 11 ） nmol/L and （ 49. 05 ±11.67 ） μmol/L respectively. Conclusion The method is simple, fast, accurate and convenient, and suitable for routine analysis.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2007年第10期1134-1137,共4页
Chinese Journal of Laboratory Medicine
关键词
色谱法
高效液相
色氨酸
犬尿喹啉酸
Chromatography, high performance liquid
Tryptophan
Kynurenic acid
