2 fév 2010 · Acid hydrolysis and Bligh and Dyer extract gave comparable extracted fatty acid contents with direct methylation The mass balance of fatty acids
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2 fév 2010 · Acid hydrolysis and Bligh and Dyer extract gave comparable extracted fatty acid contents with direct methylation The mass balance of fatty acids
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Master thesis of EMQAL project
Evaluation of Extraction Methods for Recovery
of Fatty Acids from Marine ProductsLiping Xiao 㙪Бᑇ
Supervisor: Svein Are Mjøs, Nofima Ingredients,Bjørn Grung, University of Bergen
February 2010
brought to you by COREView metadata, citation and similar papers at core.ac.ukprovided by Universidade do Algarve
Abstract
The extraction efficiency of Soxhlet, acid hydrolysis and Bligh and Dyer were evaluated by using direct methylation on extracts and residues for calculating the mass balance of fatty acids for eight marine powders (fishmeals, krillmeals, cod filet, salmon filet and herring roe). The results show that Soxhlet gave lowest extracted fatty acid content, especially for the samples which contain a high amount of phospholipid. Acid hydrolysis and Bligh and Dyer extract gave comparable extracted fatty acid contents with direct methylation. The mass balance of fatty acids in extract and residue is close to 100% for the three extraction methods which indicate that fatty acid was not lost during the extraction procedures. The difference of extracted fatty acids is mainly due to the different extracting efficiency. The gravimetric lipid has limited correlation with total fatty acids, especially forSoxhlet.
Analyses of the fatty acid profiles show ed that the Soxhlet extracts were different from the others. Extracts from the acid hydrolysis and Bligh and Dyer methods had similar fatty acid profiles as the direct methylation method. The precision of fatty acid analysis by direct methylation method for marine powders were also validated. The coefficient of variation was 5.11% for solid samples and 1.21% for liquid sample. Key words: direct methylation, one-step methylation, fatty acids, Soxhlet, acid hydrolysis, Bligh and DyerTable of Contents
List of Abbreviations.................................................................... 11 Introduction..............................................................................
21.1 Lipid nutrition in fish products.....................................................
21.2 Lipid soluble organic pollutants...................................................
21.3 Total lipid and fatty acid composition analysis................................
32 Theory ....................................................................................
52.1 Lipids...................................................................................
52.2 Fatty acids.............................................................................
52.3 Lipid classes: simple lipids and complex lipids...............................
72.4 Neutral and polar lipids............................................................
92.5 Lipid extraction principle...........................................................
102.6 Extraction methods and total lipid determination............................
112.6.1 Lipid extraction methods.......................................................
112.6.2 Commonly used methods.....................................................
122.7 Total fatty acids and fatty acid profile analysis...............................
152.7.1 Transmethylation /Methylation...............................................
152.7.2 Multistep methods vs. direct methylation methods.....................
162.7.3 GC analysis.......................................................................
172.8 Lipid class analysis.................................................................
173 Experimental Section.................................................................
193.1 Samples...............................................................................
193.2 Water content........................................................................
203.3 Extraction methods.................................................................
203.3.1 Soxhlet method..................................................................
203.3.2 Acid hydrolysis method.........................................................
223.3.3 Modified Bligh and Dyer method.............................................
233.4 T ransmethylation/ methylation method........................................
253.4.1 Preparation of methylation detergent and internal standard.........
253.4.2 Methylation procedure.........................................................
253.5 Fatty acid analysis by GC.........................................................
263.6 Lipid class analysis by LC.........................................................
273.7 Quality control........................................................................
283.8 Analysis of data......................................................................
293.9 Outline of the experiment..........................................................
294 Results and discussion..............................................................
314.1 Quality control result................................................................
324.1.1 Repeatability of fatty acid analysis by DM.................................
324.1.2 Intermediate precision for fish powder (A-I) and control oil
334.1.3 Comparison of the results of control oil................................
344.1.4 Comparison of the results of extraction methods........................
354.2 Samples...............................................................................
364.2.1 Fatty acid composition by GC................................................
364.2.2 Lipid classes by LC.............................................................
384.3 Mass balance of the extraction methods to direct methyaltion..........
394.3.1 Mass balance for the Soxhlet method......................................
404.3.2 Mass balance for the EU method............................................
414.3.3 Mass balance for the Bligh and Dyer method............................
424.3.4 Precision of the extraction methods........................................
444.4 Extracted total Fatty acids by three extraction methods...................
454.5 Gravimetric lipid content by extraction methods.............................
454.6 Influence of the extraction methods on fatty acid profiles.................
484.6.1 Introduction........................................................................
484.6.2 Fatty acid profile in extracts...................................................
494.6.3 Reconstructed fatty acid profile..............................................
514.6.4 Multivariate evaluation of the profiles.......................................
565 Conclusions............................................................................
5759
60
66
Appendix A. Data for Direct methylation (Table A1- A10) 67
Appendix B. Data for Soxhlet method (Table B1-B12) 73
Appendix C. Data for acid hydrolysis method (Table C1-C12) 85
Appendix D. Data for Bligh and Dyer method (Table D1-D12) 97
Appendix E. Data for control oil
113Appendix F. Soxhlet procedure for Nofima BioLab
115Appendix G. Acid hydrolysis procedure for Nofima BioLab 117
Appendix H. Bligh and dyer method for Nofima BioLab 119
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