Purification of a factor from human peritoneal fluid that is able to immobilize spermatozoa

Human peritoneal fluid has been claimed to influence sperm motility.This report gives evidence for the presence in mid-cycle peritonealfluid of a protein-bound, lipidic (hydrophobic) component ableto immobilize spermatozoa as a function of time. This componentwas extracted from molecular weight-sieving and ion-exchange/highpressure liquid chromatography (HPLC)-purified peritoneal fluidfractions by either chloroform/methanol or charcoal treatments;resuspension of the chloroform/methanol extract with BWW-bufferand subsequent testing on spermatozoa resulted in sperm immobilization.Sequential or step-down chromatographic procedures (molecularweight-sievingcation-exchangeanion-exchange HPLC separationsof native peritoneal fluid) and extensive dialysis against doubledistilled water allowed the purification of the sperm immobilizingfactor, as evidenced by the shorter incubation times necessaryfor sperm immobilization. Furthermore, the active fraction wasfound to immobilize spermatozoa without affecting its viability.Separation of the chloroform/methanol extracted immobilizingfraction on thin layer chromatography under conditions for phospholipiddetection allowed the identification of a characteristic bandwhich, after re-extraction, was found to be the sperm immobilizingsubstance. This factor does not contain choline, ethanolamineor serine. These results suggest that some lipidic peritonealfluid components may influence sperm motility.