Development and validation of amino acids analysis method using LC-triple-TOF and aTRAQ labeling protocol. Application in human plasma samples

V. Papaioannou; A. Yanni; V. Karathanos; M. Halabalaki

doi:10.1055/s-0041-1736846

Planta Medica, Table of Contents

Planta Med 2021; 87(15): 1270
DOI: 10.1055/s-0041-1736846

Abstracts

8. Poster Contributions

8.4 Analytics, recent methodology and applications

Development and validation of amino acids analysis method using LC-triple-TOF and aTRAQ labeling protocol. Application in human plasma samples

Authors

V. Papaioannou

¹Division of Pharmacognosy and Naturals Products Chemistry, Department of Pharmacy School of Pharmacy, National and Kapodistrian University of Athens, Greece
A. Yanni

²Laboratory of Chemistry-Biochemistry-Physical Chemistry of Foods, Department of Nutrition and Dietetics, Harokopio University of Athens, Athens, Greece
V. Karathanos

²Laboratory of Chemistry-Biochemistry-Physical Chemistry of Foods, Department of Nutrition and Dietetics, Harokopio University of Athens, Athens, Greece
M. Halabalaki

¹Division of Pharmacognosy and Naturals Products Chemistry, Department of Pharmacy School of Pharmacy, National and Kapodistrian University of Athens, Greece

Abstract

Full Text

Amino Acids (AA) analysis is of great importance in many areas of biochemical, pharmaceutical and clinical research. Currently, there are evidences that the supplementation of foods with specific AA influence appetite and energy intake especially with arginine (L-arg). The aim of this study is to develop and validate a method for absolute quantitation of AA in human plasma incorporating a triple-TOF (AB Sciex 5600⁺) platform and the aTRAQ labeling protocol as well as to demonstrate their effective compatibility. aTRAQ reagents consists of a reporter and a reactive tag that couples with the primary or secondary amine group of each AA. Ion-pair chromatography (IPC) with HFBA as ion-pair reagent was used for the separation of AA while

SWATH scan type, in positive ESI mode was employed for MS acquisition. The method was validated according to EMEA. Moreover, the newly developed method was applied in plasma samples obtained from a 12-week, two-arm parallel dietary intervention in which overweight volunteers consumed a wheat biscuit enriched with L-arg or a conventional wheat biscuitOverall, with the new method 45 AA were accurately quantified in human plasma while it was possible to monitor the L-arg plasma levels between (or in the two?) groups.