Sample preparation techniques coupled to advanced chromatographic methods for marine organisms investigation

• Folch’s, Bligh & Dyer’s and maceration for isolation of lipids from marine species.
• For derivatization of free fatty acids into methyl esters, two approaches were tested.
• Triacylglycerols analyzed by off-line Ag+-LC/NARP-LC.

Objectiveof this work was to develop suitable extraction methodologies for the isolation of lipids from fish, mussels and clams from the Mediterranean sea, and their successive analysis by means of advanced chromatographic instrumentation. More specifically, three different sample preparation methodologies were adopted: Folch’s, Bligh & Dyer’s and maceration. The lipidic extracts, after application of two different methylation procedures, were subjected to monodimensional and comprehensive two-dimensional GC analyses, in order to compare the fingerprints of samples derived from different extraction and transesterification methodologies. Triacylglycerols (TAGs) were analyzed by an off-line combination of silver-ion liquid chromatography with non-aqueous reversed phase liquid chromatography. In both LC and GC analyses, mass spectrometric detectors were used, which greatly supported the identification procedure. In particular, with respect to HPLC, mass spectrometry with atmospheric pressure chemical ionization in positive mode was applied.

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