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J Med Microbiol

Title:		High-throughput quantification of microbial-derived organic acids in mucin-rich samples via reverse phase high performance liquid chromatography
Author(s):		Villarreal AR; Lucas SK; Fletcher JR; Hunter RC;
Address:		"Department of Microbiology & Immunology, University of Minnesota, 689 23rd Avenue SE, Minneapolis, MN 55455, USA"
Journal Title:		J Med Microbiol
Year:		2023
Volume:		72
Issue:		6
Page Number:		-
DOI:		10.1099/jmm.0.001708
ISSN/ISBN:		1473-5644 (Electronic) 0022-2615 (Linking)
Abstract:		"Organic acids (short chain fatty acids, amino acids, etc.) are common metabolic byproducts of commensal bacteria of the gut and oral cavity in addition to microbiota associated with chronic infections of the airways, skin, and soft tissues. A ubiquitous characteristic of these body sites in which mucus-rich secretions often accumulate in excess, is the presence of mucins; high molecular weight (HMW), glycosylated proteins that decorate the surfaces of non-keratinized epithelia. Owing to their size, mucins complicate quantification of microbial-derived metabolites as these large glycoproteins preclude use of 1D and 2D gel approaches and can obstruct analytical chromatography columns. Standard approaches for quantification of organic acids in mucin-rich samples typically rely on laborious extractions or outsourcing to laboratories specializing in targeted metabolomics. Here we report a high-throughput sample preparation process that reduces mucin abundance and an accompanying isocratic reverse phase high performance liquid chromatography (HPLC) method that enables quantification of microbial-derived organic acids. This approach allows for accurate quantification of compounds of interest (0.01 mM - 100 mM) with minimal sample preparation, a moderate HPLC method run time, and preservation of both guard and analytical column integrity. This approach paves the way for further analyses of microbial-derived metabolites in complex clinical samples"
Keywords:		"*Mucins/metabolism Chromatography, High Pressure Liquid/methods *Respiratory System Amino Acids Fatty Acids, Volatile glycoproteins high performance liquid chromatography mucin organic acids reverse phase liquid chromatography targeted metabolomics;"
Notes:		"MedlineVillarreal, Alex R Lucas, Sarah K Fletcher, Joshua R Hunter, Ryan C eng England 2023/06/09 J Med Microbiol. 2023 Jun; 72(6). doi: 10.1099/jmm.0.001708"