LC-MS-Based Metabolic Fingerprinting of Aqueous Humor.
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, 2µL 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.
Clinical Research Centre, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Karolina
0000-0002-6639-2823
K
Pietrowska
Department of Ophthalmology, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Diana Anna
DA
Dmuchowska
Clinical Research Centre, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Paulina
0000-0001-7815-1149
P
Samczuk
Clinical Research Centre, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Tomasz
0000-0002-3979-8801
T
Kowalczyk
Department of Ophthalmology, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Pawel
P
Krasnicki
Department of Ophthalmology, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Malgorzata
M
Wojnar
Department of Ophthalmology, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Aleksandra
A
Skowronska
Department of Ophthalmology, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Zofia
Z
Mariak
Clinical Research Centre, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Adam
0000-0002-4522-4978
A
Kretowski
Clinical Research Centre, Medical University of Bialystok, M. Sklodowskiej Curie 24a, 15-276 Bialystok, Poland.
Michal
0000-0003-3729-0518
M
Ciborowski