Abstract
The aim of this study was to evaluate whether the fecal progestagen (progesterone and its metabolites) levels of miniature pigs would change after excretion at room temperature. Our initial investigation focused on the correlations between the fecal progestagen concentrations with and without ether extraction and between the plasma progesterone and fecal progestagen concentrations in order to develop an enzyme-linked immunosorbent assay (ELISA) for fecal progestagen without ether extraction. There were significant correlations between fecal progestagen concentrations with and without ether extraction (r=0.880) and between fecal progestagen concentrations without ether extraction and plasma progesterone (r=0.763). The fecal progestagen concentration obtained by ELISA without ether extraction was almost identical to that obtained with ether extraction. These results validate the ELISA method without ether extraction, which was therefore used for the latter experiment. Fecal samples collected from the pigs were preserved for 0-24 h at room temperature, and then their fecal progestagen concentrations were measured. The fecal samples preserved for 0 to 24 h were analyzed by high performance liquid chromatography (HPLC) and ELISA. The concentrations of all samples significantly increased with time after preservation. The progestagen concentration of fresh feces (0 h) with high progestagen concentration (>1000 ng/g) increased significantly after 3 h. The concentration increased significantly after 12 h for fresh feces containing about 500 ng/g progestagen. HPLC analysis is showed that the fecal progesterone concentration, but not its other metabolites, doubled 24 h after excretion compared with the concentration at 0 h. These results suggest that dynamic changes in the profile of progesterone metabolites occur in feces after excretion.