谷氨酰胺高效液相测定法的研究应用

Method for determination of glutamine in serum

目的 建立一种准确、简便、快速的高效液相色谱法 ,快速测定血中谷氨酰胺含量 ,监测给予经谷氨酰胺强化的安全肠外营养病人血清谷氨酰胺浓度并与对照组比较。方法 反相C1 8柱为固定相 ,异硫氰酸苯酯作柱前衍生剂 ,5 0mmol L醋酸缓冲液 (pH 6 .5 0 ) 乙腈 (97.7∶2 .3)为流动相 ,进行单泵等梯度洗脱 ,在λ2 5 4nm处检测谷氨酰胺含量。结果 线性范围 40～ 2 5 0 0 μmol L ,相关系数0 .998,批内和批间的变异系数分别为 2 .5 6 %和 2 .85 % ,高低两种浓度的回收率分别为 (10 3.2±2 .9) %和 (96 .8± 2 .1) % ,临床健康人血清谷氨酰胺浓度参考值为 (6 16 .8± 6 8 0 ) μmol L ,手术后经谷氨酰胺强化的完全肠外营养患者血清谷氨酰胺浓度明显高于对照组 (P <0 .0 5 )。结论 本法简便、快速 ,无需梯度洗脱 ,分离效果好 ,适用于临床有关血清谷氨酰胺的检测和研究。

Objective To establish a new high performance liquid chromatography method for determining the concentration of glutamine(GLN) in human serum and to observe the change of concentration of GLn in post surgical patients by using glutamine enriched parenteral nutrition.Methods GLN were derivatized with phenylisothiocyanate(PITC) and eluted with 50 mmol/L sodium acetate(pH6.5)containing 23% acetonitrile on reverse phase C 18 column. The derivative was detected at 254 nm.Results Calibration curves for GLN was linear from 40 to 2 500 μmol/L( r =0.998), and CV was between 2.56% and 2.85%. The mean recovery was 99.5%, the normal values in health adults were (616.8±68.0) μmol/L, and the concentration of GLN was higher for patients after operation than for conrols.Conclusion This method is simple, rapid and suitable for clinical and basic stodies.