Simultaneous Determination of Multiple Amino Acids in Plasma in Critical Illness by High Performance Liquid Chromatography with Ultraviolet and Fluorescence Detection

Overview

Journal J Chromatogr B Analyt Technol Biomed Life Sci

Publisher Elsevier

Specialties Biomedical Engineering
Chemistry

Date 2013 Oct 23

PMID 24145014

Citations 12

Authors

Hao Wang

Yvette R McNeil

Tsin W Yeo

Nicholas M Anstey

Affiliations

Soon will be listed here.

Abstract

There is increasing recognition that the host response to critical illness includes derangement of multiple amino acid pathways, including amino acids (AAs) central to metabolism and immune, endothelial and neurological function. To characterise concentration changes of these plasma amino acid we report the development and validation of a method for the quantification of AAs in small volumes of plasma (50μL) using HPLC with simultaneous UV and fluorescence (FL) detection. Protein precipitation and pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is followed by reversed phase HPLC separation. Calibration curves were built with norleucine as an internal standard. Thirty-three (including the 20 proteinogenic) AAs, were selected as standards and their corresponding concentrations in the plasma of healthy human controls and patients with severe falciparum malaria were quantified. This method enables the detection of perturbations in arginine metabolism, aromatic amino acid pathways, methionine transsulfuration and transmethylation pathways and other metabolic pathways.

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