Thiele Björn, Hupert Michelle, Santiago-Schübel Beatrix, Oldiges Marco, Hofmann Diana
Institute of Bio- and Geosciences, IBG-2: Plant Sciences, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany.
Institute of Bio- and Geosciences, IBG-3: Agrosphere, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany.
Methods Mol Biol. 2019;2030:403-414. doi: 10.1007/978-1-4939-9639-1_30.
In this chapter we describe a method for quantification of 20 proteinogenic amino acids by liquid chromatography-mass spectrometry which affords neither derivatization nor the use of organic solvents. Analysis of the underivatized amino acids is performed by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) in the positive ESI mode. Separation is achieved on a strong cation exchange (SCX) column (Luna 5 μ SCX 100 Å) with 5% acetic acid in water (A) and 75 mM ammonium acetate in water (B). Quantification is accomplished by use of d-phenylalanine as internal standard achieving limits of detection of 5-50 nM. The method was successfully applied for the determination of proteinogenic amino acids in plant extracts.
在本章中，我们描述了一种通过液相色谱 - 质谱法对20种蛋白质氨基酸进行定量的方法，该方法既无需衍生化，也无需使用有机溶剂。未衍生化氨基酸的分析通过液相色谱 - 电喷雾电离 - 串联质谱法（LC - ESI - MS/MS）在正ESI模式下进行。在强阳离子交换（SCX）柱（Luna 5 μ SCX 100 Å）上，以5%乙酸水溶液（A）和75 mM乙酸铵水溶液（B）实现分离。通过使用d - 苯丙氨酸作为内标进行定量，检测限达到5 - 50 nM。该方法已成功应用于植物提取物中蛋白质氨基酸的测定。
