Comparison of Eleven Extraction Methods for Quantitative Evaluation of Total Lipids and Fatty Acids in the Clam Anadara inaequivalvis (Bruguière)

The clam Anadara inaequivalvis (Bruguière) is a bivalve mollusc now populating several areas of the Mediterranean Sea seafloor where it competes with other molluscs harvested as food. In fact, even though Anadara inaequivalvis has valuable nutritional properties, it has no commercial value in European countries. The aim of this study was to evaluate quantitatively and qualitatively fatty acid content in this bivalve mollusc and to compare 11 extraction methods: conventional procedures such as Folch, Bligh and Dyer, and Soxhlet with different solvents, as well as accelerated solvent extraction (ASE) and methods using low-toxicity solvents as substitutes for chloroform (CHCl3) and methanol (MeOH). The best method was ASE CHCl3-MeOH, affording yields of total lipids, total fatty acids, and polyunsaturated fatty acids (PUFAs) significantly higher than any other method. The estimated range of total lipids was 2.29%–6.99% (on the dry sample), and the range of fatty acids was 0.74%–1.78%. Yields of polyunsaturated fatty acids (PUFAs) ranged from 0.10% to 0.40%. The solvent mixture hexane–isopropanol as substitute for CHCl3-MeOH did not afford good results. PUFAs were 22.5% in the total fatty acids and the ratio between PUFAs n-3 and PUFA n-6 was 2.74:1.