Background: The aim of this study was to evaluate the integrity of sperm surface characteristics in the presence of a new male contraceptive, RISUG [1 mg styrene maleic anhydride (SMA)/100 μl dimethylsulphoxide (DMSO) in 1 ml sperm solution]. Methods: Progressively motile human sperm were treated in vitro with RISUG. The cells were analysed for the release of 5′-nucleotidase (5′-NT) (a plasma membrane marker) using 3 mmol/l 5′-AMP and 3 mmol/l β-glycerophosphate as substrates. Hyaluronidase (an acrosomal membrane marker) was analysed using hyaluronic acid as a substrate. The contents of free and total acrosin, and % proacrosin (all acrosome markers) were assayed using 0.5 mmol/l α -N-benzoyl-L-arginine ethylester (BAEE). Results: RISUG caused almost complete disintegration of the plasma membrane leading to significant (P < 0.0001) release of 5′-NT into the surrounding media. Complete dissolution of the acrosome with concomitant vesiculation of the membrane system, as judged from the loss of hyaluronidase, was observed. Total acrosin content in the sperm was also reduced to almost 10%, and proacrosin dropped to 13.2% in the presence of RISUG in comparison to 90.2% in control (P < 0.0001), indicating dispersion of acrosomal contents. Conclusion: Under in vitro conditions, RISUG, at a concentration of 1 mg SMA dissolved in 100 μl of DMSO, caused significant damage to the acrosome and its contents, indicating loss of functional ability of sperm. © European Society of Human Reproduction and Embryology 2004; all rights reserved.