The relationship between plasma levels of total phospholipids (PL) and/or PL fractions and neoplastic diseases are not fully understood. Therefore, the aim of this study was to analyze concentrations and distribution of plasma phospholipids in patients with prostate cancer (PCa) related to the Gleason score, clinical stage and pathologic grade of prostate cancer. We analyzed plasma phospholipids in 57 newly diagnosed, untreated PCa patients and in 43 age-matched healthy male subjects. Significantly lower (P < 0.01) levels of total plasma PL and all PL classes were found in PCa patients when compared with healthy subjects. The relative concentrations of PL fractions were also changed. Further decrease of total PL and PL fractions was found related to an increase of clinical stadium, pathologic grade, and Gleason score, with phosphatidylethanolamine as the most sensitive plasma PL, the level of which significantly decreased even at the first stage of PCa. Our results showed an altered plasma PL profile in PCa patients, which may contribute to monitoring of the disease progression.