高血压患者血浆中游离氨基酸的柱前衍生HPLC法测定

Determination of Free Plasma Amino Acids in Hypertensive Patient by Pre-column Derivatization HPLC

以2,4-二硝基氯苯为衍生化试剂,建立了反相高效液相色谱法测定人血浆中22种游离氨基酸的方法。衍生化产物用Kromasil C18柱分离,梯度洗脱,紫外检测波长为360nm。在所研究的浓度范围内,22种氨基酸的线性关系良好(r>0.99),日内精密度(RSD)为1.7%～4.1%,日间精密度(RSD)为2.8%～6.9%,方法回收率为90%～106%。用建立的方法对高血压病人和正常人血浆中的游离氨基酸进行测定,并将测定结果进行主成分分析及判别分析,结果表明高血压病人与正常人血浆中的氨基酸存在显著差异,分类判别率为94.4%。

A reversed phase high performance liquid chromatographic （RP-HPLC） method for simulta- neous determination of 22 free plasma amino acids was established by using 2, 4-dinitrochlorobenzene as pre-column derivatization agent. The amino acid derivatives were separated on a Kromasil C18 column with gradient elution mode and detected at wavelength of 360 nm. The calibration curves showed linearity over the studied concentration range. The intra-day and inter-day preeisions were 1.7%- 4. 1% and 2. 8%-6.9% , respectively. The spiked recoveries were between 90% and 106% . The method was applied in the determination of the concentration of the free plasma amino acids in hyper- tensive patients and healthy subjects. The experimental data were further analyzed by the principal component and classification method, which indicated that there was a clear differentiation between healthy people and hypertension patients with the classification value of 94. 4%.