ETHNOPHARMACOLOGICAL RELEVANCE: Pedalium murex Linn. has been used as Vajikaran Rasayana (aphrodisiac) in traditional Indian medicine to treat male sexual dysfunction and impotency. AIM OF THE STUDY: The aim of this study was to investigate effects of P. murex fruits extract on sexual behaviors and testosterone level of male rats during and past withdrawal of treatment. MATERIALS AND METHODS: The extract (50, 100,150mg/kg body weight/day) and sildenafil citrate (5mg/kg body weight/day) were administered orally by gavages for 28 days to male Wistar albino rats. Penile erection index (PEI), mount latency (ML), intromission latency (IL), ejaculation latency (EL), mounting frequency (MF), intromission frequency (IF), post ejaculatory interval (PEjI) and serum testosterone levels were studied at day 0, 15, 28 during treatment. They were further evaluated after day 7 and 15 past discontinuation of the treatment. In-vitro nitric oxide release activity was also investigated in human corpus cavernosal cell line. RESULTS: The ethanolic extract significantly reduced the ML, IL, EL and PEjI (p<0.05). There was a significant increase in the PEI, MF and IF and serum testosterone level (p<0.05) throughout the period of study. Ethanolic extract produced a significant effect on sexual behavior and serum testosterone level past withdrawal of the treatment. In-vitro nitric oxide release was significantly higher in extract and sildenafil citrate compared to the control group. CONCLUSIONS: Present findings provide experimental in-vivo and in-vitro evidence that the ethanolic extract of P. murex fruits possesses aphrodisiac property. Study lends growing support for the traditional use of P. murex as a sexual stimulating agent and offers a significant potential for studying the effect on male sexual response and its dysfunctions. The findings justify the concept of Rasayana as rejuvenative tonics and support their role in prevention or delay of the aging process.