滤纸上血斑中苯丙氨酸和酪氨酸的高效液相色谱测定法。

HPLC assay of phenylalanine and tyrosine in blood spots on filter paper.

作者信息

Roesel R A, Blankenship P R, Hommes F A

出版信息

Clin Chim Acta. 1986 Apr 15;156(1):91-6. doi: 10.1016/0009-8981(86)90183-x.

Abstract

Blood spots on filter paper are used to screen for phenylketonuria. We have developed a high pressure liquid chromatographic assay of phenylalanine and tyrosine after elution of these amino acids from filter paper. The analysis time is 20 min. The amino acids are separated using an ion-exchange resin with post-column ortho-phthalaldehyde derivatization and subsequent fluorescence detection. Other common amino acids do not interfere. The assay is linear from 20 pmol to at least 2,000 pmol with a coefficient of variation of 3%. The assay is as accurate as the determination of phenylalanine by ion-exchange ninhydrin-reactive methods currently in use, but is 100 times more sensitive. The method has the advantages of avoiding venipuncture, is cost effective, has a high sensitivity, and a rapid turnaround.

摘要

滤纸上的血斑用于筛查苯丙酮尿症。我们开发了一种高压液相色谱法，用于在从滤纸洗脱这些氨基酸后测定苯丙氨酸和酪氨酸。分析时间为20分钟。使用离子交换树脂进行氨基酸分离，柱后邻苯二甲醛衍生化并随后进行荧光检测。其他常见氨基酸不产生干扰。该测定法在20皮摩尔至至少2000皮摩尔范围内呈线性，变异系数为3%。该测定法与目前使用的离子交换茚三酮反应法测定苯丙氨酸一样准确，但灵敏度高100倍。该方法具有避免静脉穿刺、成本效益高、灵敏度高和周转迅速的优点。