Application of liquid chromatographymass spectrometry to measure the concentrations and study the synthesis of short chain fatty acids following stable isotope infusions
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abstract
A new method involving zinc sulphate deproteinization was developed to study short chain fatty acids (SCFA) production in the colon and subsequent occurrence of SCFA in blood. SCFA were baseline separated in a 30min cycle using ion-exclusion chromatography and detected by mass spectrometry. Concentrations could be measured down to 10M and isotopomeric distributions could be assessed, enabling the conduction of tracer studies to study changes in SCFA synthesis. The applicability of the method was tested in an extensively characterized pig model yielding portal SCFA concentrations ranging from 70M (butyric acid) to 150M (propionic acid) to 440M (acetic acid) prior to butyrate tracer infusion, reaching butyric acid isotopic steady state within 2h.