Glutamate and GABA concentrations in feces and colonic contents were determined by high performance liquid chromatography (HPLC) coupled with a photodiode-array (PDA) detector (HPLC-PDA) on fecal waters following diethyl ethoxymethylenemalonate (DEEMM, Sigma-Aldrich) derivatization according to previously described procedures (19 (link)). First, samples were weighed and diluted with 20 volumes of PBS, using Stomacher for homogenization. Then, free amines and amino acids were first extracted using acidification with hydrochloric acid and filtrated using 3-kDa cutoff Amicon filtering units (Millipore-Merck). Finally, extracted samples were subjected to DEEMM derivatization and analyzed by HPLC-PDA as previously described.
Differences between male and female animals were first assessed statistically using two-way ANOVA, and when no differences were detected, treatment groups including both genders were compared using one-way ANOVA.
Differences between male and female animals were first assessed statistically using two-way ANOVA, and when no differences were detected, treatment groups including both genders were compared using one-way ANOVA.
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