Back to Search Start Over

NMR-Based Identification of Metabolites in Polar and Non-Polar Extracts of Avian Liver

Authors :
Fariba Fathi
Antonio Brun
Katherine H. Rott
Paulo Falco Cobra
Marco Tonelli
Hamid R. Eghbalnia
Enrique Caviedes-Vidal
William H. Karasov
John L. Markley
Source :
Metabolites, Vol 7, Iss 4, p 61 (2017)
Publication Year :
2017
Publisher :
MDPI AG, 2017.

Abstract

Metabolites present in liver provide important clues regarding the physiological state of an organism. The aim of this work was to evaluate a protocol for high-throughput NMR-based analysis of polar and non-polar metabolites from a small quantity of liver tissue. We extracted the tissue with a methanol/chloroform/water mixture and isolated the polar metabolites from the methanol/water layer and the non-polar metabolites from the chloroform layer. Following drying, we re-solubilized the fractions for analysis with a 600 MHz NMR spectrometer equipped with a 1.7 mm cryogenic probe. In order to evaluate the feasibility of this protocol for metabolomics studies, we analyzed the metabolic profile of livers from house sparrow (Passer domesticus) nestlings raised on two different diets: livers from 10 nestlings raised on a high protein diet (HP) for 4 d and livers from 12 nestlings raised on the HP diet for 3 d and then switched to a high carbohydrate diet (HC) for 1 d. The protocol enabled the detection of 52 polar and nine non-polar metabolites in 1H NMR spectra of the extracts. We analyzed the lipophilic metabolites by one-way ANOVA to assess statistically significant concentration differences between the two groups. The results of our studies demonstrate that the protocol described here can be exploited for high-throughput screening of small quantities of liver tissue (approx. 100 mg wet mass) obtainable from small animals.

Details

Language :
English
ISSN :
22181989
Volume :
7
Issue :
4
Database :
Directory of Open Access Journals
Journal :
Metabolites
Publication Type :
Academic Journal
Accession number :
edsdoj.10ae90776d904005adf1c925ff89f333
Document Type :
article
Full Text :
https://doi.org/10.3390/metabo7040061