The present work evaluates the use of cardoon (Cynara cardunculus L.) seed oil as a renewable raw material for biodiesel production using an endogenous plant of the Mediterranean Flora. Cardoon seed oil was extracted using supercritical carbon dioxide (scCO(2)) with different densities. Depending on the employed extraction conditions and/or methods, different oil yields and compositions were obtained. As expected, higher scCO(2) densities led to higher cardoon oil yields similar to the Soxhlet one. Two biodiesel synthesis reactions were performed to convert oil esters into fatty acid methyl esters: alkali-based transesterification and acid esterification followed by alkali-based transesterification. All the obtained biodiesel samples were chemically and physically characterized. Results showed that the two-step transesterification led to higher biodiesel yields as well as higher fatty acid methyl ester (FAME) contents. Moreover, other measured properties fulfilled, with a few exceptions, the specified EN14214 values. (C) 2012 Elsevier B.V. All rights reserved.