The present invention relates to a novel biological fuel oil, in particular to a biological oil using citrus oil as a material and a preparation method thereof. The materials used by the technology are the offal and rejects of fruits, making a break through in the prior production mode of biological diesel oil at home and abroad, which is the biggest contribution made by the technology. The citrus oil is regenerative energy, and can substitute petrifaction fuel. Citrus fruit is evergreen xylophyta which is beneficial to ecological environment improvement. The method for preparing the biological fuel oil by using the citrus oil as the material comprises the following steps that: that raw oil extracted from the materials which are pericarps, stones, roots, branches and leaves, etc. of rutaceae citrus plants such as lemon, pomelo, orange, king orange, mandarin, etc. is subjected to water filtrating, impurity filtering and substance such as pigment, etc. extraction so as to obtain the oil; the oil mainly contains limonene of 95 percent, which is terpene with a molecular formula C10H16 and can be directly used as fuel oil to ignite and burn. The biological oil also can be prepared from the citrus oil of between 50 and 80 percent and methanol (ethanol) of between 20 and 50 percent, by weight percentage.