Purpose: Proton magnetic resonance spectroscopy (1 H-MRS) has been proposed as a tool to assess male infertility providing metabolic signatures related to the spermatogenesis in the testis. This study sought to identify the role of 1 H-MRS in the diagnosis of infertility in patients with semen analysis altered. Materials and Methods: 14 patients (27 testicles) with fertility problems and with an altered semen analysis (5 oligospermia, 3 asthenospermia, 6 oligoasthenospermia) and 9 controls (18 testicles) with normal spermatogenesis assessed (men with prior paternity and normal semen analysis) and normal testicles at magnetic resonance (MR) and ultrasonography (US) with colour Doppler (CD) examination were included. All patients underwent testis US and CD investigation, conventional MR examination at 1.5T including T1 and T2 weighted images in three orthogonal planes and proton magnetic resonance spectroscopy (1 H-MRS) with single-voxel PRESS (TR 2000 ms / TE 31 ms). Major metabolites peaks (choline, creatine, lipids, lactate) were calculated and compared between the patients and controls. Results: Mean choline peak in the semen analysis altered group was statistically significantly lower than the normal group (0.69 vs 1.34, 95% CI: 0.52 - 0.85; p < 0.001). 18 testicles within semen analysis altered group (66,7%) had both MR and US examination negative but mean choline peak lower then controls (1.09 vs 1.34, p < 0.001). 7 testicles of those presented also varicocele at CD investigation. Conclusion: 1 H-MRS revealed a significant shift towards lower choline peak in patients with semen analysis altered compared to controls with normal spermatogenesis. Moreover 1 HMRS provided to find out spermatogenesis disorder in patients with normal testis at MR and US examination.