Terrlink T, van Leeuwen P A, Houdijk A
Department of Clinical Chemistry, Free University Hospital, Amsterdam, The Netherlands.
Clin Chem. 1994 Feb;40(2):245-9.
We present an HPLC method for the determination of amino acids in plasma. The method is based on automated precolumn derivatization of amino acids with o-phthalaldehyde, separation of the derivatives by reversed-phase chromatography, and quantification by fluorescence detection. Complete separation was achieved within 12 min. Total analysis time, including derivatization, chromatography, and reequilibration of the column, was 17 min. The assay was linear from 5 to 800 mumol/L for all amino acids. Recovery of amino acids added to plasma samples was 96-106%, except for tryptophan (89%). Within-run precision (CV) was 1.8-6.4%, and between-run precision was 2.1-7.2%. The method can be used for determining primary amino acids in plasma and cerebrospinal fluid. The simple sample preparation and short analysis time make the method particularly suitable for routine analysis of large series of samples.
我们提出了一种用于测定血浆中氨基酸的高效液相色谱法。该方法基于氨基酸与邻苯二甲醛的自动柱前衍生化、通过反相色谱法分离衍生物以及通过荧光检测进行定量。在12分钟内实现了完全分离。包括衍生化、色谱分析和柱再平衡在内的总分析时间为17分钟。所有氨基酸的测定线性范围为5至800μmol/L。添加到血浆样品中的氨基酸回收率为96 - 106%,色氨酸除外(89%)。批内精密度(CV)为1.8 - 6.4%,批间精密度为2.1 - 7.2%。该方法可用于测定血浆和脑脊液中的伯氨基酸。简单的样品制备和较短的分析时间使该方法特别适用于大量样品的常规分析。