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Journal of Analytical Methods in Chemistry
Volume 2017, Article ID 6745932, 13 pages
https://doi.org/10.1155/2017/6745932
Research Article

LC-MS-Based Metabolic Fingerprinting of Aqueous Humor

1Clinical Research Centre, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland
2Department of Ophthalmology, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland

Correspondence should be addressed to Michal Ciborowski; lp.ude.bmu@iksworobic.lahcim

Received 7 September 2016; Accepted 6 December 2016; Published 5 January 2017

Academic Editor: Josep Esteve-Romero

Copyright © 2017 Karolina Pietrowska et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Aqueous humor (AH) is a transparent fluid which fills the anterior and posterior chambers of the eye. It supplies nutrients and removes metabolic waste from avascular tissues in the eye. Proper homeostasis of AH is required to maintain adequate intraocular pressure as well as optical and refractive properties of the eye. Application of metabolomics to study human AH may improve knowledge about the molecular mechanisms of eye diseases. Until now, global analysis of metabolites in AH has been mainly performed using NMR. Among the analytical platforms used in metabolomics, LC-MS allows for the highest metabolome coverage. The aim of this study was to develop a method for extraction and analysis of AH metabolites by LC-QTOF-MS. Different protocols for AH preparation were tested. The best results were obtained when one volume of AH was mixed with one volume of methanol : ethanol (1 : 1). In the final method, 2 µL of extracted sample was analyzed by LC-QTOF-MS. The method allowed for reproducible measurement of over 1000 metabolic features. Almost 250 metabolites were identified in AH and assigned to 47 metabolic pathways. This method is suitable to study the potential role of amino acids, lipids, oxidative stress, or microbial metabolites in development of ocular diseases.