柱前衍生高效液相色谱法测定人血浆中17种氨基酸的含量

Determination of 17 Amino Acids in Human Plasma by High Performance Liquid Chromatography with Pre-column Derivatization

目的应用6-氨基喹啉-N-羟基琥珀酰亚胺基氨基甲酸酯(AQC)柱前衍生高效液相色谱法测定正常人血浆中17种氨基酸的含量。方法血浆样品与AQC衍生,衍生产物用氨基酸专用C18色谱柱进行分离,梯度洗脱,使用荧光检测器(λex=250 nm,λem=395 nm)。用建立的方法对正常人血浆中的17种游离氨基酸进行测定(n=22)。结果在所研究的浓度范围内,17种氨基酸线性关系良好(R2>0.99),平均回收率在(87.7±5.8)%与(117.1±8.4)%之间,批内和批间变异系数分别在10.3%和11.0%以下。正常人血浆中17种氨基酸测定结果均在正常参考值范围内。结论该方法简便、稳定、分离效果好,可用于人血浆中氨基酸浓度的检测。

Objective To detect 17 amino acids in human plasma by HPLC with pre-column derivatization using 6-amin- oquinolyl-N-hydroxysuccinimidyl carbamate （AQC）. Methods Plasma samples were pretreated with AQC as derivatization a- gent. The amino acid derivatives were separated on a C18 column with fluorescence detection （kex=250 nm, λem= 395nm） after gradient elution. The method was applied to determine the concentration of 17 free amino acids in human plasma from 22 healthy w）lunteers. Results The analytes displayed a good linearity over the tested concentration range. The average recover7 was within the range of （87.7±5.8） % - （ 117.1 ±8.4） %. The intra- and inter-variation coefficients were less than 10.3% and 11.0%, re- spectively. All tested amino acids in the plasma samples are within the normal reference range. Conclusion The method is sim- ple and stable with decent separation, which can be generally used to determine amino acids in human plasma.