Figure 1. Purification of a bioactive substance from utero-vaginal junction extracts.
(a) Utero-vaginal junction (UVJ) extracts were loaded onto a Superdex 200 pg column. The detectable peak was collected and each fraction was evaporated to dryness, dissolved in a physiological saline (pH 7.4), and applied to the sperm motility assay. The bioactive fraction is indicated by the horizontal bar. (b) HPLC profile of bioactive fractions (in panel a) on a C-22 reverse-phase column. Each collected fraction is indicated with a horizontal bar and a numeral. (c) Effects of HPLC fractions on sperm motility. Ejaculated sperm were suspended in a medium containing each fraction (1% (v/v)) and the effects of each fraction on sperm motility were evaluated. Bioactivity was detected in fractions 2 and 3. Asterisk indicates a significant difference from H2O. (d) Thin layer chromatography (TLC) analysis of the bioactive substance. A mixture of fractions 2 and 3 (in panel b) was further separated through preparative TLC. The motility score for sperm in each fraction are indicated below the TLC photograph. White arrow indicates the bioactive substance. S, mixture of fractions 2 and 3.