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通过超高效液相色谱-紫外分析对复杂生物样品中AccQ-Tag衍生化氨基酸和生物胺进行快速灵敏定量分析。

Fast and Sensitive Quantification of AccQ-Tag Derivatized Amino Acids and Biogenic Amines by UHPLC-UV Analysis from Complex Biological Samples.

作者信息

Guba Andrea, Bába Orsolya, Tőzsér József, Csősz Éva, Kalló Gergő

机构信息

Proteomics Core Facility, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Egyetem tér 1, 4032 Debrecen, Hungary.

Biomarker Research Group, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Egyetem tér 1, 4032 Debrecen, Hungary.

出版信息

Metabolites. 2022 Mar 21;12(3):272. doi: 10.3390/metabo12030272.

Abstract

Metabolomic analysis of different body fluids bears high importance in medical sciences. Our aim was to develop and validate a fast UHPLC-UV method for the analysis of 33 amino acids and biogenic amines from complex biological samples. AccQ-Tag derivatization was conducted on target molecules and the derivatized targets were analyzed by UHPLC-UV. The detection of the analytes was carried out with UV analysis and by Selected Reaction Monitoring (SRM)-based targeted mass spectrometry. The method was validated according to the FDA guidelines. Serum and non-stimulated tear samples were collected from five healthy individuals and the samples were analyzed by the method. The method was successfully validated with appropriate accuracy and precision for all 33 biomolecules. A total of 29 analytes were detected in serum samples and 26 of them were quantified. In the tears, 30 amino acids and biogenic amines were identified and 20 of them were quantified. The developed and validated UHPLC-UV method enables the fast and precise analysis of amino acids and biogenic amines from complex biological samples.

摘要

不同体液的代谢组学分析在医学领域具有重要意义。我们的目标是开发并验证一种快速超高效液相色谱 - 紫外检测(UHPLC - UV)方法,用于分析复杂生物样品中的33种氨基酸和生物胺。对目标分子进行AccQ - Tag衍生化,然后通过UHPLC - UV对衍生化后的目标物进行分析。使用紫外分析和基于选择反应监测(SRM)的靶向质谱法进行分析物的检测。该方法根据美国食品药品监督管理局(FDA)指南进行了验证。从五名健康个体采集血清和非刺激泪液样本,并采用该方法进行分析。该方法对所有33种生物分子均成功验证,具有适当的准确度和精密度。在血清样本中共检测到29种分析物,其中26种被定量。在泪液中,鉴定出30种氨基酸和生物胺,其中20种被定量。所开发并验证的UHPLC - UV方法能够对复杂生物样品中的氨基酸和生物胺进行快速、精确的分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f84c/8949038/c6d663de0c17/metabolites-12-00272-g001.jpg

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