Location by paper chromatography of compensatory ovarian hypertrophy (COH) inhibiting activity in acetic acid extracts from bovine pineals

Acetic acid extracts of bovine pineals and cerebral cortex were separated on Sephadex G-25 columns. Subsequently two low molecular weight fractions, F₂ and F₃, were ultrafiltered through the membranes UM2 and UM05. The UM05 residues were gel filtered on Sephadex G-15 columns or chromatographed on Dowex W50-X4 columns. Fractions from these columns were tested and those which showed COH-inhibiting activity were separated by preparative paper chromatography in different solvents. The absorption spectra of those fractions were recorded and tested for COH-inhibition. By these methods, a COH-inhibitor was localized in three different solvents. Some active paper chromatography fractions were studied in high pressure, reverse phase, liquid chromatography. This latter method showed that the active fractions obtained by paper chromatography contain several orthophthalaldehyde (OPT) positive compounds.