Sherwood R A, Titheradge A C, Richards D A
Biochemistry Department, Royal Sussex County Hospital, Brighton, East Sussex, U.K.
J Chromatogr. 1990 Jun 29;528(2):293-303. doi: 10.1016/s0378-4347(00)82388-9.
The use of reversed-phase liquid chromatography (LC) with pre-column derivatization for the analysis of amino acid mixtures is becoming established as a possible cheaper alternative to commercial amino acid analysers. The available derivatization procedures all have disadvantages when applied to clinical samples, partly due to the interferences found with body fluids when ultraviolet or fluorescence detection is used. An LC method is described for the separation of amino acids in blood or urine, using pre-column derivatization with phenylisothiocyanate (PITC), gradient elution and electrochemical detection. The use of electrochemical detection of PITC derivatives virtually eliminates interferences and enables the secondary amino acids to be measured. Examples are shown of normal urine and plasma and samples from patients with cystinuria and maple syrup urine disease.
采用柱前衍生化反相液相色谱法（LC）分析氨基酸混合物，正逐渐成为一种比商用氨基酸分析仪更经济的可行替代方法。现有的衍生化方法应用于临床样本时均存在缺点，部分原因是使用紫外或荧光检测时会受到体液干扰。本文描述了一种液相色谱法，用于分离血液或尿液中的氨基酸，该方法采用异硫氰酸苯酯（PITC）进行柱前衍生化、梯度洗脱和电化学检测。对PITC衍生物进行电化学检测几乎消除了干扰，并能够测定仲氨基酸。文中给出了正常尿液和血浆以及胱氨酸尿症和枫糖尿症患者样本的示例。
