| Affiliation: | 1. Marine Bioprospecting Section of Marine Biotechnology Division, Central Marine Fisheries Research Institute, Cochin, India;2. Department of Biosciences, Mangalore University, Mangalagangothri, Karnataka State, India |
| Abstract: | The C20-22 polyunsaturated fatty acids (C20-22 PUFAs), mainly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), were concentrated from the refined liver oil of deep sea leafscale gulper shark, Centrophorus squamosus, by sequential processes of winterization, urea complexation, and argentation chromatography. Winterization at 4°C using acetonitrile as solvent showed significant reduction of the total saturated fatty acid content (12%) with a concomitant increase of polyunsaturated fatty acids (36.3%). The urea complexation process significantly enriched the total polyunsaturated fatty acids (49.5%), with a reduction in saturated fatty acids (83.9%). Silica gel based argentation chromatography further concentrated the esters of C20-22 PUFA (>99 percent purity). Nuclear Magnetic Resonance spectroscopy was used as a chemical fingerprinting tool to study the enrichment of C20-22 PUFAs at various stages of the purification process. An increase in the proton signal intensity at the olefinic region (at δ 4–6) and a decrease in the aliphatic signals (at δ 0.5–2) showed that the process successfully enriched the C20-22 PUFAs. Abbreviations: PUFA, polyunsaturated fatty acid; EPA, eicosapentaenoic acid; DHA, docosahexaenoic acid; FAME, fatty acid methyl esters; SFA, saturated fatty acid; MUFA, monounsaturated fatty acid; NMR, nuclear magnetic resonance; GCMS, gas chromatography mass spectrometry; FFA, free fatty acid. |
