This article reports biodiesel production and discusses experimental results from a laboratory performance study by using natural novel source of non edible wild safflower ( Carthamus oxyacantha Bieb.) oil. The effect of variables including molar ratio, reaction temperature, reaction time, concentration of catalyst, and mixing intensity on biodiesel yield and characteristics of the biodiesel produced from wild safflower oil were analyzed. The highest triglyceride conversion rate of 84% was achieved after 2 hr of reaction at 65°C, with a 5:1 molar ratio of methanol to oil and sodium hydroxide as catalyst. It is stated that the wild safflower oil biodiesel (WSOB) is comparable with the recommended ASTM biodiesel standard and can be used as option for biomass energy.