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United States Department of Agriculture

Agricultural Research Service

Research Project: IMPACT OF EARLY DIETARY FACTORS ON CHILD DEVELOPMENT AND HEALTH

Location: Arkansas Children's Nutrition Center

Title: Lipid fatty acid profile analyses in liver and serum in rats with nonalcoholic steatohepatitis using improved gas chromatography-mass spectrometry methodology

Authors
item Wu, Xianli -
item Tong, Yudong -
item Shankar, Kartik -
item Baumgardner, January -
item Kang, Jie -
item Badeaux, Jamie -
item Badger, Thomas
item Ronis, Martin -

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 30, 2010
Publication Date: January 26, 2011
Citation: Wu, X., Tong, Y., Shankar, K., Baumgardner, J.N., Kang, J., Badeaux, J., Badger, T.M., Ronis, M.J. 2011. Lipid fatty acid profile analyses in liver and serum in rats with nonalcoholic steatohepatitis using improved gas chromatography-mass spectrometry methodology. Journal of Agricultural and Food Chemistry. 59(2):747-754.

Interpretive Summary: Fatty acids (FA) are essential components of lipids and exhibit important biological functions. In this study, the GC-MS methodology, which was routinely used to analyze FA, was improved. This new method was utilized to analyze free fatty acids, and the FA profiles of triglycerides and phospholipids in the serum and liver, from a recently established rat model of nonalcoholic steatohepatitis (NASH). The results indicated that alteration of FA composition in liver, after overfeeding of high polyunsaturated fat diets, may contribute to the progression of pathological changes from steatosis to inflammation, necrosis, and fibrosis observed in NASH.

Technical Abstract: Fatty acids (FA) are essential components of lipids and exhibit important biological functions. The analyses of FAs are routinely carried out by gas chromatography-mass spectrometry, after multi-step sample preparation. In this study, several key experimental factors were carefully examined, validated, and optimized to analyze free fatty acid (FFA) and FA profiles of triglycerides and phospholipids in serum or tissue samples. These factors included: 1) methylation/ transeserification reagents; 2) validation of internal standards; 3) final step concentration of FA methyl esters. This new method was utilized to analyze FFAs and the FA profiles of triglycerides and phospholipids in the serum and liver, from a recently established rat model of nonalcoholic steatohepatitis (NASH). In this model, rats were fed a 220 kcal.kg -3/4 day -1 diet, containing either 5% or 70% corn oil for 21 day using total enteral nutrition. FA composition of the serum and liver, were found to shift from a pattern dominated by saturated and monounsaturated FAs (C16:0/18:1), to one dominated by polyunsaturated C18:2 derived from dietary linoleic acid. Alteration of FA composition in liver, after overfeeding of high polyunsaturated fat diets, may contribute to the progression of pathological changes from steatosis to inflammation, necrosis, and fibrosis observed in NASH.

Last Modified: 8/22/2014