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In recent years, alternative sources of fuel are receiving a lot of attention in the automotive industry. Fuels derived from an agricultural feedstock are an attractive option. Bio-fuels based on vegetable oils offer the advantage being a sustainable, annually renewable source of automobile fuel. One of key issues in using vegetable oil based fuels is its oxidation stability. Since diesel fuels from fossil oil have good oxidation stability, automobile companies have not considered fuel degradation when developing diesel engines and vehicles as compared with gasoline engines. This paper presents the results of oxidation stability testing on bio-fuels. Oxidation stability was determined using three test methods, ASTM D525, EN14112 and ASTM D2274. The effects of storage condition, bio-fuel composition and antioxidants on the degradation of bio-fuels were all investigated. ASTM D525 is an effective test method to determine the effects of storage condition on bio-fuels stability.