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A Closer Examination of 6-Aminoquinolyl-N-Hydroxysuccinimidyl Carbamate Amino Acid Derivatization in HPLC with Multiple Detection Modes
Chromatographia ( IF 1.7 ) Pub Date : 2021-06-04 , DOI: 10.1007/s10337-021-04051-w
Yu-Sheng Sung , Alain Berthod , Daipayan Roy , Daniel W. Armstrong

6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate is a well-known reagent used to derivatize amino acids (AAs) into their aminoquinolyl carbamate (AQC) analogs. Originally, AQC derivatization was used to enhance UV detection of amino acids (and amines) and to provide a fluorescent derivative for further enhanced detection. Subsequently, it was found that AQC derivatization could also enhance mass spectrometry (MS) detection. It is sometimes assumed that since the AQC tag is identical for all amino acids, that their detection will be similar for a given detection mode. However, this is often not the case. Sometimes there are extreme exceptions, for example, cysteine and tryptophan, which are simply ignored in the literature. Herein, four detection modes were compared: two optical spectroscopic modes, 254 nm UV and fluorimetry, and two mass spectrometry (MS) modes, single ion analysis and multiple reaction monitoring (MRM). The most sensitive detection modes were fluorimetry and MRM which were able to detect down to the tens of femtomoles of injected AAs. But single quad MS and classical UV detection could quantitate most AQC-AAs down to the picomole level in most instances. The differences between detection modes and within each of the four detection modes are examined and discussed for the first time. The possible reasons for extreme differences in the detectability of specific amino acids are considered.



中文翻译:

在具有多种检测模式的 HPLC 中对 6-氨基喹啉基-N-羟基琥珀酰亚胺氨基甲酸酯氨基酸衍生化的进一步研究

6-氨基喹啉-N-羟基琥珀酰亚胺氨基甲酸酯是一种众所周知的试剂,用于将氨基酸 (AA) 衍生为它们的氨基喹啉氨基甲酸酯 (AQC) 类似物。最初,AQC 衍生化用于增强氨基酸(和胺)的紫外检测,并提供荧光衍生物以进一步增强检测。随后,发现 AQC 衍生化还可以增强质谱 (MS) 检测。有时假设由于所有氨基酸的 AQC 标签都是相同的,因此对于给定的检测模式,它们的检测将是相似的。然而,情况往往并非如此。有时也有极端的例外,例如半胱氨酸和色氨酸,它们在文献中被简单地忽略了。在本文中,比较了四种检测模式:两种光谱模式,254 nm 紫外和荧光法,以及两种质谱 (MS) 模式,单离子分析和多反应监测 (MRM)。最灵敏的检测模式是荧光法和 MRM,它们能够检测到低至数十飞摩尔的注射 AA。但在大多数情况下,单四极杆 MS 和经典 UV 检测可以将大多数 AQC-AA 定量至皮摩尔水平。首次检查和讨论了检测模式之间以及四种检测模式中的每一种之间的差异。考虑了特定氨基酸可检测性存在极端差异的可能原因。但在大多数情况下,单四极杆 MS 和经典 UV 检测可以将大多数 AQC-AA 定量至皮摩尔水平。首次检查和讨论了检测模式之间以及四种检测模式中的每一种之间的差异。考虑了特定氨基酸的可检测性存在极端差异的可能原因。但在大多数情况下,单四极杆 MS 和经典 UV 检测可以将大多数 AQC-AA 定量至皮摩尔水平。首次检查和讨论了检测模式之间以及四种检测模式中的每一种之间的差异。考虑了特定氨基酸的可检测性存在极端差异的可能原因。

更新日期:2021-06-04
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