Kato Megumi, Takatsu Akiko
Bio-Medical Standard Group, National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki, Japan.
Methods Mol Biol. 2019;2030:111-118. doi: 10.1007/978-1-4939-9639-1_9.
Here, we describe an amino acid analysis that is based on the use of hydrophilic interaction liquid chromatography coupled with isotope dilution mass spectrometry for the accurate quantification of underivatized amino acids from hydrolyzed peptide/protein. Twelve underivatized amino acids were separated and detected during an 88-min runtime. The absolute limits of detection and limits of quantification (on column) of the four amino acids (isoleucine, phenylalanine, proline, and valine) were in the range of 6-80 and 20-200 fmol, respectively. As little as 25 pmol of peptide or protein hydrolysate is sufficient for determining absolute content.
在此，我们描述了一种氨基酸分析方法，该方法基于亲水作用液相色谱与同位素稀释质谱联用，用于准确量化水解肽/蛋白质中的未衍生化氨基酸。在88分钟的运行时间内分离并检测了12种未衍生化氨基酸。四种氨基酸（异亮氨酸、苯丙氨酸、脯氨酸和缬氨酸）的绝对检测限和定量限（柱上）分别在6 - 80飞摩尔和20 - 200飞摩尔范围内。仅25皮摩尔的肽或蛋白质水解产物就足以测定绝对含量。
