A streamlined approach to the analysis of volatile fatty acids and its application to the measurement of whole-body flux.

Volatile fatty acids (VFAs) are produced in the human colon by the bacterial breakdown of carbohydrates that escape digestion and absorption in the small intestine. They have important local and systemic effects on gastrointestinal and nutritional functions. Measuring their production is difficult because of inaccessibility of sampling sites and low circulating concentrations. Stable isotope tracer techniques are a way to measure VFA production but require measurement of isotope dilution in blood and other biological fluids. We have developed a streamlined and robust method to measure the concentration and enrichment of [(2)H]-labelled VFAs by gas chromatography/mass spectrometry (GC/MS) and [(13)C]-labelled VFAs by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). Both types of analysis were carried out on the same samples allowing multiple tracer studies to be conducted. Good accuracy and repeatability were found for GC/MS analysis of [(2)H]-labelled VFAs. Careful handling of the background contribution, especially acetate, allowed quantitation of concentration and enrichment within the analysis. GC/C/IRMS analysis of [(13)C] VFAs was also achieved with good accuracy and repeatability. This methodology was used to determine whole-body acetate production in two subjects using multiple tracers ([(2)H(3)]- and [1-(13)C]acetate) and blood and urine sampling. Whole-body acetate flux was similar when measured either with [(2)H(3)]- or [1-(13)C]acetate, and when flux was determined from plasma or urine tracer enrichment. This new method will permit rapid and accurate measurement of VFA flux using [(2)H]- and/or [(13)C]-labelled VFAs as tracers. Measurements of the contribution of colonic VFA production to whole-body VFA flux are now possible.