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Coumarin-based derivatization reagent for LC-MS analysis of amino acids
Talanta ( IF 6.1 ) Pub Date : 2022-08-05 , DOI: 10.1016/j.talanta.2022.123730
Nguyễn Kim Ngân Bùi 1 , Sigrid Selberg 1 , Koit Herodes 1 , Ivo Leito 1
Affiliation  

In this paper, we report about the application of a sensitive fluorescent derivatization reagent Coumarin151-N-Hydroxysuccinimidyl Carbamate (Cou151DSC) for amino compounds using high-performance liquid chromatography (HPLC) compatible with ultraviolet (UV), fluorescence detector (FLD) and electrospray ionization - tandem mass spectrometry (ESI-MS/MS)―positive mode. We optimized derivatization procedure and validated an analytical method to determine 24 amino acids in Kvass drink using Norvaline as an internal standard. Compared to 6-Aminoquinolyl-N-Hydroxysuccinimidyl Carbamate (6-AQC), the derivatization with Cou151 DSC is faster and milder, for 5 min at 40°C instead of 15 min at 55°C. The limit of quantitation (LOQ, pmol on column) for 21 amino acids in this work is lower 1.1–30.0 times than values obtained with 6-AQC. The derivatives have excitation wavelength at 355 nm and emission wavelength at 486 nm. Their MS/MS fragmentation behaviors were examined together with 23 other amino compounds. We found three possibilities to lose a neutral group which can be Coumarin 151 isocyanate Cou151NCO (255 Da), amine Coumarin 151 (229 Da) or urea Cou151CONH2 (272 Da). The accuracy of the proposed method was within 83–107% with good relative standard deviations (RSDs) of equal or less than 6%. The recoveries were from 82 to 120% in four spiked concentrations, repeatability was between 0 and 14%. The intra- and inter-day precision are less than 13% and 18%, respectively.



中文翻译:

用于氨基酸 LC-MS 分析的基于香豆素的衍生化试剂

在本文中,我们使用与紫外 (UV)、荧光检测器 (FLD) 和电喷雾兼容的高效液相色谱 (HPLC)报告了灵敏的荧光衍生试剂 Coumarin151- N -Hydroxysuccinimidyl 氨基甲酸酯 (Cou151DSC) 在氨基化合物中的应用电离 - 串联质谱 (ESI-MS/MS)——正离子模式。我们优化了衍生化程序并验证了一种分析方法,以使用正缬氨酸作为内标来测定格瓦斯饮料中的 24 种氨基酸。与 6-氨基喹啉-N相比-羟基琥珀酰亚胺氨基甲酸酯 (6-AQC),使用 Cou151 DSC 的衍生化更快更温和,在 40°C 下进行 5 分钟而不是在 55°C 下进行 15 分钟。本研究中 21 种氨基酸的定量限(LOQ,色谱柱上的 pmol)比 6-AQC 获得的值低 1.1-30.0 倍。衍生物的激发波长为 355 nm,发射波长为 486 nm。他们的 MS/MS 碎裂行为与其他 23 种氨基化合物一起进行了检查。我们发现了三种失去中性基团的可能性,可以是香豆素 151 异氰酸酯 Cou151NCO (255 Da)、胺香豆素 151 (229 Da) 或尿素 Cou151CONH 2(272 大)。所提出方法的准确度在 83-107% 之间,相对标准偏差 (RSD) 良好,等于或小于 6%。四个加标浓度的回收率为 82% 至 120%,重复性为 0 至 14%。日内和日间精密度分别低于 13% 和 18%。

更新日期:2022-08-05
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