The aim of this study was to evaluate the vitamin levels of ram seminal plasma and the relationship between vitamins and spermatological characteristics. For this aim, nine 3-5 year old rams of Norduz, Karakas and Ile-de-France x Akkaraman (G1) genotypes were provided. Sperm of each ram was collected once per month from June to October of the year 2007 by artificial vagina. In each month, the concentrations of retinol acetate, alpha tocopherol, delta tocopherol, alpha tocopherol acetate and vitamin D-3 in seminal plasma were analyzed by using HPLC. Spermatological characteristics viz. sperm volume, pH, mass activity, sperm motility, abnormal spermatozoid, death/alive ratio and sperm concentration were evaluated. As testicular characteristics, scrotal circumference and testicular diameter were measured. Monthly changes and breed differences on the vitamin levels were statistically insignificant, whereas there was a positive correlation was obtained between testis diameter and retinol acetate levels (p<0.05) and a positive correlation was also estimated between analyzed vitamins. As a result, the relationships between the testicular, spermatological characteristics, and vitamin compositions should be taken into account in the breeding targets.