LAPORAN HASIL PENELITIAN
PROGRAM RESEARCH GRANT I-MHERE JURUSAN
TAHUN ANGGARAN 2010
Judul : Sintesis Fosfolipid Mengandung Asam Lemak ω-3
dari Fosfolipid Kedelai dan Minyak Kaya Asam Lemak ω -3 dari Hasil Samping Pengalengan Tuna Ketua : Mokhamad Nur, STP, MSc.
Anggota : Dr. Teti Estiasih, STP, MP
Dibiayai Oleh Direktorat Jenderal Pendidikan Tinggi, Kementerian Pendidikan Nasional, melalui DIPA Universitas Brawijaya berdasarkan SK Rektor Nomor :
214/SK/2010, tanggal 22 Juli 2010.
UNIVERSITAS BRAWIJAYA
NOPEMBER
2010
RINGKASAN
Fosfolipid merupakan pengemulsi yang bersifat alami. Fosfolipid komersial yang biasa digunakan oleh industri pangan adalah fosfolipid atau lesitin kedelai. Keunggulan fosfolipid dapat lebih ditingkatkan dengan menggabungkan asam lemak ω-3, terutama EPA (eicosapentaenoic acid, C20:5 ω-3) dan DHA docosahexaenoic acid, C22:6 ω-3), pada struktur fosfolipid sehingga diperoleh fosfolipid terstruktur. Fosfolipid kedelai mengandung asam lemak ω-3 merupakan inovasi baru bahan penyedia asam lemak ω-3 sekaligus juga sebagai pengemulsi.
Asam lemak ω-3 merupakan asam lemak yang penting bagi kesehatan dengan sumber utama adalah minyak ikan. Minyak ikan hasil samping pengalengan tuna merupakan sumber asam lemak ω-3 yang potensial dengan keunggulan tinggi asam dokosaheksaenoat. Kadar asam lemak ω-3 dalam minyak ikan dapat ditingkatkan dengan menggunakan teknik kristalisasi pelarut suhu rendah. Pada proses sintesis fosfolipid terstruktur diperlukan sumber asil tinggi asam lemak ω-3. Pada penelitian ini, sumber asil yang digunakan adalah minyak kaya asam lemak ω-3 dalam bentuk asam lemak bebas yang diperoleh dari teknik kristalisasi pelarut suhu rendah. Kristalisasi pelarut suhu rendah dilakukan setelah minyak hasil samping pengalengan tuna disaponifikasi dan diekstrak asam lemaknya. Asam lemak jenuh dengan titik leleh tinggi akan mengkristal sedangkan asam lemak ω-3 karena mempunyai titik leleh rendah diharapkan tetap cair sehingga dapat dipisahkan dengan asam lemak jenuh.
Strukturisasi fosfolipid dilakukan dengan cara mengganti asam lemak dari fosfolipid kedelai dengan asam lemak ω-3 dari minyak kaya asam lemak ω-3 dari hasil samping pengelangen tuna yang diperoleh dengan teknik kristalisasi pelarut suhu rendah. Sintesisi fosfolipid terstruktur dilakukan secara asidolisis enzimatis dengan menggunakan lipase dari R.miehei. Faktor yang dikaji pada proses sintesis ini adalah konsentrasi enzim dan lama sintesis. Variabel respon yang diteliti adalah tingkat inkorporasi asam lemak ω-3 (EPA+DHA) pada fosfolipid terstruktur dan nilai HLB. Fosfolipid terstruktur mengandung asam lemak ω-3 yang dihasilkan dengan tingkat inkorporasi EPA+DHA tertinggi selanjutnya dikarakterisasi meliputi profil fosfolipid dan tingkat inkorporasi EPA+DHA pada jenis-jenis fosfolipid.
Hasil penelitian menunjukkan bahwa teknik kristalisasi pelarut suhu rendah dengan menggunakan asam lemak bebas hasil hidrolisis sebagai sumber minyak dapat meningkatkan kadar EPA+DHA sebanyak 1,37 kali. Tingkat inkorporasi EPA dan DHA pada struktur fosfolipid dipengaruhi oleh konsentrasi enzim dan lama reaksi asidolisis enzimatis. Ada interaksi antara konsentrasi enzim dan lama reaksi. Pada konsentrasi enzim yang rendah, peningkatan lama reaksi setelah tingkat inkoporasi optimum tercapai cenderung menyebabkan penurunan tingkat inkorporasi EPA+DHA. Pada konsentrasi enzim yang tinggi, lama reaksi tampaknya tidak mempengaruhi tingkat inkorporasi.
Tingkat inkorporasi DHA lebih tinggi dari EPA sehingga fosfolipid terstruktur yang dihasilkan sangat sesuai digunakan untuk produk pangan yang memerlukan kadar DHA tinggi seperti makanan bayi. Kemampuan emulsifikasi fosfolipid terstruktur yang ditunjukkan oleh nilai EAI dan ESI dipengaruhi oleh tingkat inkorporasi EPA+DHA. Ada preferensi inkorporasi EPA+DHA pada fosfatidiletanolamin dibandingkan jenis fosfolipid yang lain.
Kata kunci: fosfolipid, fosfolipid terstruktur, asam lemak ω-3, asidolisis enzimatis
DAFTAR PUSTAKA
Adlercreutz, D. and E. Wehtje. 2004. An enzymatic method for the synthesis of mixed-acid phosphatidylcholine. J. Am. Oil Chem. Soc. 81: 553-557.
Ahmadi, K. 2006. Optimasi Kristalisasi Pelarut Suhu Rendah pada Pembuatan Minyak Kaya Asam Lemak -3 dari Hasil Samping Pengalengan Ikan Lemuru (Sardinella longiceps). Laporan Penelitian Dosen Muda.
Aïd, S., S. Vancassel, A. Linard, M. Lavialle, and P. Guesnet. 2005. Dietary docosahexaenoic acid [22: 6(n-3)] as a phospholipid or a triglyceride enhances the potassium chloride–evoked release of acetylcholine in rat hippocampus. J. Nutr. 135:1008-1013.
Ajina, M.Z, A. Gharib, G. Durand, N. Gazzah, B. Claustrat, C. Gharib, and N. Sarda. 1999. Dietary docosahexaenoic acid-enriched phospholipids normalize urinary melatonin excretion in adult (n-3) polyunsaturated fatty acid-deficient rats. J. Nutr. 129:2074-2080.
Amate, L., A. Gil, and M. Ramírez. 2001. Feeding infant piglets formula with long-chain polyunsaturated fatty acids as triacylglycerols or phospholipids influences the distribution of these fatty acids in plasma lipoprotein fractions. J. Nutr. 131:1250-1255.
AOAC. 1996. Official Methods of Analysis. 16th ed. Vol 2. Virginia, USA.
Aura, A-M., P. Forsell, A. Mustranta, and K. Poutanen. 1995. Transesterification of soy lecithin by lipase and phospholipase. J. Am. Oil Chem. Soc. 72: 1375-1379.
Calder, P.C. 2004. n-3 fatty acids, inflammatory, and immunity relevance to post-surgical and critically Ill patients. Lipids 39: 1147-1161.
Chen, T-C. and Y-H. Ju. 2001. Polyunsaturated fatty acids concentrates from borage and linseed oil fatty acids. J. Am. Oil Chem. Soc. 78(5): 485-488.
Colbert, L.B. 1998. Lecithins tailored to your emulsification needs. INFORM 43(9): 686-688.
Coste, T.C., A. Gerbi, P. Vague, G. Pieroni, and D. Raccah. 2003. Neuroprotective Effect of docosahexaenoic acid–enriched phospholipids in experimental diabetic neuropathy. Diabetes 52:2578-2585.
Crawford, M.A., M. Bloom, C.I. Broadhurst, W.F. Schmidt, S.C. Cunnane, C. Galli, K. Gehbremeskel, F. Linseisen, J. Llyod-Smith, and J. Parkington. 1999. Evidence for the unique function of docosahexaenoic acid during the evolution of the modern hominid brain. Lipids 34: S39-S45..
Dashiell, G.L. 1989. Lecithin in Food Processing Applications. In B.F. Szuhaj (ed.). Lecithins: Sources, Manufacture, and Uses. The American Oil Chemists’ Society, Champaign, Illinois.
Erjawetz, S.P, J. Ulrich, M. Tiedtke and R.W Hartel. 1999. Milk fat fractionation by solid-layer melt crystallization. J. Am. Oil Chem. Soc. 76(5): 579-584.
Estiasih, T. 2003. Peran Natrium Kaseinat dan Fosfolipid dalam Emulsifikasi dan Mikroenkapsulasi Trigliserida Kaya Asam Lemak ω-3. Disertasi. Program Pascasarjana, Universitas Gadjah Mada, Yogyakarta.
Estiasih, T., K, Ahmadi, dan W.B. Sunarharum, 2009a. Pengembangan Ekstraksi dan Saponifikasi Satu Tahap pada Pembuatan Konsentrat PUFA dan Protein secara Simultan dari Kedelai Varietas Lokal. Laporan Hibah Penelitian Kompetitif. LPPM UB, Malang.
Estiasih, T., K. Ahmadi, dan F.C. Nisa. 2009. Sintesis Pengemulsi Mengandung Asam Lemak -3 dari Hasil Samping Pengolahan Ikan dan Kelapa Sawit. Laporan Tahun II, Insentif Riset Dasar.
Estiasih, T., K. Ahmadi, dan F.C. Nisa. 2009b. Sintesis Pengemulsi Mengandung Asam Lemak -3 dari Hasil Samping Pengolahan Ikan dan Kelapa Sawit. Laporan Tahun II, Insentif Riset Dasar. LPPM UB, Malang.
Fennema, O.R. 1996. Food Chemistry. 3rd Edition. Marcel Dekker, Inc., New York.
Garcia, H. S., I. Kim, A.L. Hernandez, and C.G. Hill. 2008. Enrichment of licithin with n-3 fatty acids by acidolysis using immobilized phospholipase A1. Grasas Y Aceities 59(4) : 368-374.
Garg, M.L., L.G. Wood, H. Singh, and P.J. Moughan. 2006. Means of delivering recommended levels of long chain n-3 polyunsaturated fatty acids in human diets. J. Food Sci. 71(5): R66.
Haagsma. N., Vant Gent, C. M. Luten, J. B., de Jong, R. W. and Van Doorn, E. 1982. Preparation of an ω-3 fatty acid concentrate from cod liver oil. J. of Amer. Oil Chem. Soc. 59(3): 117–118.
Hamama, A.A. and H.L. Bhardwaj. 2004. Phytosterol, triterpene alcohols, and phospholipids in seed oil from white lupin. J. Amer. Oil Chem. Soc. 81(11): 1039-1044.
Hara, F., T. Nakashima, and H. Fukuda. 1997. Comparative study of commercially available lipases in hydrolysis reaction of phosphatidylcholine. J. Am. Oil Chem. Soc. 74: 1129–1132.
Haraldsson, G.G. and A. Thorarensen. 1999. Preparation of phospholipids highly enriched with n-3 polyunsaturated fatty acids by lipase. J. Am. Oil Chem. Soc. 76(10): 1143-1149.
Hardman, W.E. 2002. Omega-3 fatty acids to augment cancer therapy. J. Nutrition 132:3508S-3512S.
Hardman, W.E. 2004. (n-3) fatty acids and cancer terapi. J. Nutrition 134: 3427S-3430S.
Hayes, D.G. 2004. Enzyme-catalyzed modification of oilseed materials to produce eco-friendly products. J. of Amer. Oil Chem. Soc. 81: 1077-1103.
Hosokawa, M., K. Takahashi, N. Miyazaki, K. Okamura, and M. Hatano. 1995. Application of water mimics on preparation of eicosapentaenoic and docosahexaenoic acid containing glycerolipids. J. Am. Oil Chem. Soc. 72: 421-425.
Howe, P.R.C., J.A. Downing, B.F.S. Grenyer, E.M. Grigonis-Deane, and W.L. Bryden. 2002. Tuna fishmeal as a source of DHA for n-3 PUFA enrichment of pork, chicken, and eggs. Lipids 37: 1067-1076.
Huang, M.T., G. Ghai, and C.T. Ho. 2004. Inflammatory process and molecular targets for anti-inflammatory nutraceuticals. Comprehensive Reviews in Food Science and Food Safety 3: 127-139.
Ikrawan, Yusep. 2004. Minyak Ikan dan Omega 3. http://www.pikiran-rakyat.com. Diakses 20 Januari 2008.
Irianto, H. E. dan Indroyono S.. 2007. Dukungan Teknologi Penyediaan Produk Perikanan. http:// www.litbang.deptan.go.id/. Diakses tanggal 3 Februari 2008.
Ishinaga, M., A. Ueda, T. Mochizuki, S. Sugiyama, and T. Kobayashi. 2005. Cholesterol intake is associated with lecithin intake in Japanese people. J. Nutr. 135:1451-1455.
Jolly, C.A., A. Muthukumar, C.P.R. Avula, D. Troyer, and G. Fernandes. 2001. Autoimmune-prone (NZBXNZW) (1) mice fed a diet enriched with (n-3) fatty acids. J. Nutrition 131: 2753-2760.
Joshi, A., S.G. Paratkar, and B.N. Thorat. 2006. Modification of lecithin by physical, chemical and enzymatic methods. Eur. J. Lipid Sci. Technol. 108 (2006) 363– 373
Judde, A., P. Villeneuve, A. Rossignol-Casteraa, and A.L Guillou. 2003. Antioxidant effect of soy lecithins on vegetable oil stability and their synergism with tocopherols. J. Am. Oil Chem. Soc. 80: 1209–1215.
Kinsella, J.E. 1986. Food components with potential therapeutic benefits in the n-3 polyunsaturated fatty acids of fish oils. Food Technology, February
Krishnamurthy, R.and M.Kellens. 1995. Fractionation and winterization. In Y-H.Hui (ed.). Bailey’s Industrial Oil and Fat Products. Edible Oil and Fat Products: Processing Technology. 5th ed. Vol. 4. A John Wiley & Sons, Inc., New York
Leitzmann, M.F., M.J. Stampfer, D.S. Michaud, K. Augustsson, G.C. Colditz, W.C. Willet, and E.L. Giovarnucci. 2004. Dietary intake of n-3 & n-6 fatty acids and the risk of prostate cancer. Am. J. Clin. Nutr. 80(1): 204-216.
Lyberg, A-M., E. Fasoli, and P. Adlercreutz. 2005. Monitoring the oxidation of docosahexaenoic acid in lipids. J. Am. Oil Chem. Soc. 40: 969-979.
Mead, J.F., R.B. Alfin-Slater, D.R. Howton, and G. Popjak. 1986. Lipids. Chemistry, Biochemistry, and Nutrition. Plenum Press, New York.
Mine, Y., M. Shimizu, and T. Nakashima. 1997. Microfltration and stabilization of egg yolk phospholipid emulsion by a microporous glass membrane. J. Amer. Oil Chem. Soc. 74: 1255-1258.
Mitchell, D.C., S-L. Niu, and B.J. Litman. 2003. DHA-rich phospholipids optimize C-protein–coupled signaling. J. Pediatr. 143: 880-886.
Moffat, C. F., McGill, A. S., Hardy, R. and Anderson, R. S. 1993. The production of fish oil enriched in polyunsaturated fatty acid-containing triglycerides. J. of Amer. Oil Chem. Soc. 70 (2): 133-138.
Mori, T.A., D.Q. Bao, V. Burke, I.E. Puddey, G.F. Watts, and L.J. Beilin. 1999. Dietary fish as a major component of a weight-loss diet: effect on serum lipids, glucose, and insulin metabolism in overweight hypertensive subjects. Am. J. Clin, Nutr. 70(5): 817-825.
Mustranta, A., T.Suorti, and K. Poutanen, 1994. Transesterification of phospholipids in different reaction conditions. J. Am. Oil Chem. Soc. 71: 1415-1419.
Nasir, M.I., M. A. Bernards, and P. A. Charpentier . 2007. Acetylation of soybean lecithin and identification of components for solubility in supercritical carbon dioxide. J. Agric. Food Chem. 55: 1961-1969.
Nettleton, J.A. 1995. Omega-3 Fatty Acids and Health. Chapman & Hall, New York.
Nettleton, J.A. 2005. Omega-3 fatty acids in food and health. Food Tech. 59:120.
Nwosu, C.V., L. Boyd, and B. Sheldon. 1997. Effect of fatty acid composition of phospholipids on their antioxidant properties and activity index. J. Amer. Oil Chem. Soc. 74: 293-297.
Nzai, J.M. and A. Proctor. 1998. Phospholipids determination in vegetable oil by thin layer chromatography and imaging densitometry. Food Chem. 63: 571-576.
Park, P.W. and R.E. Goins. 1994. In situ preparation of fatty acids methyl ester for analysis of fatty acids composition in foods. J. Food Sci. 59: 1262-1266.
Pearce, K.N. and J.E. Kinsella. 1978. Emulsifying properties of proteins: evaluation of a turbidimetric technique. J. Agric. Food Chem. 26(3): 716-723.
Reddy, J.R.C., T. Vijeeta, M.S.L. Karuna, B.V.S.K. Ra, and R.B.N. Prasad. 2005. Lipase-catalyzed preparation of palmitic and stearic acid-rich phosphatidylcholine. J. 74: 1531-1536. Am. Oil Chem. Soc. 82: 727-730.
Rotsstein, N.P., M.I. Avedalno, and L.E. Politi. 1999. Essentiality of docosahexaenoic acid in retina photoreceptor cell development. Lipids 34: S115.
Singleton, J.A., M. Ruan, J.H. Sanford, C.A. Haney, and L.F. Stikeleather. 1999. Separation and characterization of peanut phospholipids molecular species using high performance liquid chromatography and fast bombardment mass spectrometry. J. Am. Oil Chem. Soc. 76: 49-56.
Song, J.K., J.J. Han. And J.S. Rhee. 2005. Phospholipases: occurrence and production in microorganism, assay for high-troughput screening, and gene discovery from natrural and man-made diversity. J. Am. Oil Chem. Soc. 82: 691-705.
Spector, A.A. 1999. Essentiality of fatty acids. lipids 34: s1-s3.
Taylor, L.A., L. Pletschen, J. Arends, C. Unger, and U. Massing. 2009. Marine phospholipids—a promising new dietary approach to tumor-associated weight loss. Support Care Cancer. Published online 29 April 2009.
Terry, P.D., T.E. Rohan, and A. Wolk. 2003. Intakes of fish and marine fatty acids and the risks of cancer of the breast and prostate and of other hormone-related cancers: A review of the epidemiologic evidence. Am. J. Clin. Nutr. 77(3): 532-543.
Tiemeier, H., H.R. van Tuije, A. Hofman, A.J.Kilian, and M.M.B. Breteler. 2003. Plasma fatty Acid composition and depression are associated in the elderly: the Rotterdam study. Am. J. Clin. Nutr. 78(1): 40-46.
Uauy, R. and D.R. Hoffman. 2000. Essential fat requirements of preterm infants. Am. J. Clin. Nutr. 71(1): 245S-250S.
Verlengia, R., R. Gorjao, C.C. Kanunfre, S. Bordin, T.M. deLima, E.F. Martins, P. Newsholme, and R. Curi. 2004. Effects of EPA and DHA on proliferation cytokine production and gene expression in Raji cells. Lipids 39(9): 857-864.
Vikbjerg, A.F., H. Mu, and X. Xu. 2005b. Monitoring of monooctanoylphosphatidylcholine synthesis by enzymatic acidolysis between soybean phosphatidylcholine and caprylic acid by thin-layer chromatography with a flame ionization detector. Agric. Food Chem. 53: 3937-3942.
Vikbjerg, A.F., H. Mu, and X. Xu. 2006. Elucidation of acyl migration during lipase-catalyzed production of structured phospholipids. J. Am. Oil Chem. Soc. 83: 609–614.
Vikbjerg, A.F., J-Y. Rusig, G. Jonsson, H. Mu, and X. Xu. 2006. Comparative evaluation of the emulsifying properties of phosphatidylcholine after enzymatic acyl modification. J. Agric. Food Chem. 2006,
Vikbjerg, A.F., L. Peng, H.Mu, and X.Xu. 2005a. Continuous production of structured phospholipids on a packed bed reactor with lipase from Thermomyces lanuginosa. J. Am. Oil Chem. Soc. 82: 237-242.
Wang, G. and T. Wang. 2008. Oxidative stability of egg and soy lecithin as affected by transition metal ions and pH in emulsion. J. Agric. Food Chem. 56 (23): 11424-11431.
Wu, Y. and T. Wang. 2004. Fractionation of crude soybean lecithin with aqueous ethanol. J. Am. Oil Chem. Soc. 81: 697–704.
Zhang, F. and A. Proctor. 1997. Rheology and stability of phospholipid-stabilized emulsions. J. Am. Oil Chem. Soc. 74: 869–874.
