柱前衍生反相高效液相法定量检测烧伤患者血清氨基酸的含量

The Quantitative Determination of Serum Amino Acids in Burn Patients by Pre-column Derivatization Reversed-Phase HPLC

目的:应用柱前衍生的高效液相色谱法,定量测定重症烧伤病人血清中氨基酸的含量。方法:以2、4-二硝基氟苯(DNFB)为衍生剂,用反相高效液相色谱法在360 mm处测定血清中17种游离氨基酸的含量,并以外标法进行计算。结果:在27分内分出17种氨基酸,分离完全。DNFB衍生的氨基酸其回收率在97.32％～104.26％之间,变异系数为1.5％～4.8％。在重症烧伤病人组,血清中多种氨基酸含量发生显著改变。结论:本法操作简便,快速准确,重复性好,适合于临床检测的需要。

Objective:To quantitatively determine the content of serum amino acids in patients with severe burn by pre-column de-rivatization reversed-phase high performance liquid chromatography (HPLC). Methods;Using 2,4-dinitrofluorobanzene (2, 4-DNFB) as a derivative agent, the contents of 17 kinds of free amino acid at 360 nm were detected by reversed-phase HPLC and calculated by external standard method. Results:The 17 kinds of free amino acid derived from DNFB were completely separated in 27 minutes. Their rates of recovery varied from 97.32% to 104.26% and coefficients of variation were 1.5 % ~ 4.8 % . The obvious changes of content of many amino acids occurred in patients with severe burn. Conclusion: The reversed-phase HPLC is a simple,rapid and exact method suitable to clinical detection.