Use of Gas Chromatography to Quantify Short Chain Fatty Acids in the Serum, Colonic Luminal Content and Feces of Mice

Author(s)

Ribeiro, Willian Rodrigues ^[1] ; Ramirez Vinolo, Marco Aurelio ^[2] ; Calixto, Leandro Augusto ^[3] ; Ferreira, Caroline Marcantonio ^[3]

Affiliation

^[1] Univ Fed Sao Paulo, Chem Biol Grad Program, Diadema, SP - Brazil

^[2] Univ Estadual Campinas, Inst Biol, Lab Immunoinflammat, Campinas, SP - Brazil

^[3] Univ Fed Sao Paulo, Inst Environm Chem & Pharmaceut Sci, Dept Pharmaceut Sci, Diadema, SP - Brazil

• Document type: Journal article
• Source: BIO-PROTOCOL; v. 8, n. 22 NOV 20 2018.
• Web of Science Citations: 1

Abstract

Short-Chain Fatty Acids (SCFAs) are a product of the fermentation of resistant starches and dietary fibers by the gut microbiota. The most important SCFA are acetate (C2), propionate (C3) and butyrate (C4). These metabolites are formed and absorbed in the colon and then transported through the hepatic vein to the liver. SCFAs are more concentrated in the intestinal lumen than in the serum. Butyrate is largely consumed in the gut epithelium, propionate in the liver and acetate in the periphery. SCFAs act on many cells including components of the immune system and epithelial cells by two main mechanisms: activation of G-protein coupled receptors (GPCRs) and inhibition of histone deacetylase. Considering the association between changes in SCFA concentrations and the development of diseases, methods to quantify these acids in different biological samples are important. In this study, we describe a protocol using gas chromatography to quantify SCFAs in the serum, feces and colonic luminal content. Separation of compounds was performed using a DB-23 column (60 m x 0.25 mm internal diameter {[}i.d.]) coated with a 0.15 mu m thick layer of 80.2% 1-methylnaphatalene. This method has a good linear range (15-10,000 mu g/ml). The precision (relative standard deviation {[}RSD]) is less than 15.0% and the accuracy (error relative {[}ER]) is within +/- 15.0%. The extraction efficiency was higher than 97.0%. Therefore, this is cost effective and reproducible method for SCFA measurement in feces and serum. (AU)

• FAPESP's process: 12/50410-8 - Effects of short-chain fatty acids produced by probiotic bacteria in the prophylaxis and treatment of allergic airway inflammation
• Grantee: Caroline Marcantonio Ferreira
• Support Opportunities: Research Grants - Young Investigators Grants