Nina Skall Nielsen

Oxidative Stability of Marine Phospholipids in the Liposomal Form and Their Applications

Marine phospholipids (MPL) have attracted a great deal of attention recently as they are considered to have a better bioavailability, a better resistance towards oxidation and a higher content of eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) than oily triglycerides (fish oil) from the same source. Due to their tight intermolecular packing conformation at the sn-2 position and their synergism with α-tocopherol present in MPL extracts, they can form stable liposomes which are attractive ingredients for food or feed applications. However, MPL are still susceptible to oxidation as they contain large amounts polyunsaturated fatty acids and application of MPL in food and aquaculture industries is therefore a great challenge for researchers. Hence, knowledge on the oxidative stability of MPL and the behavior of MPL in food and feed systems is an important issue. For this reason, this review was undertaken to provide the industry and academia with an overview of (1) the stability of MPL in different forms and their potential as liposomal material, and (2) the current applications and future prospects of MPL in both food and aquaculture industries with special emphasis on MPL in the liposomal form.

Purification and deodorization of structured lipids by short path distillation

Purification of structured lipids (SL), produced from lipase-catalyzed acidolysis of rapeseed oil and capric acid, and deodorization of randomized SL, produced from chemical randomization of fish oil and tricaprin, were studied in a bench-scale short path distillation (SPD). SL obtained from enzymatic acidolysis usually contain a large proportion of medium-chain and long-chain free fatty acids. Two SPD steps have been applied for the removal of free fatty acids. Parameters such as evaporator temperature, feeding flow rate, stirring roller speed, and the content of free fatty acids (FFA) added to the starting materials were optimized with respect to FFA left in the product residuals and to tocopherol loss from the starting oil. Evaporator temperature and flow rate were optimized using response surface methodology and two models were obtained for the FFA content left and loss of tocopherols. An applicable parameter zone was created to obtain a certain FFA (0.5% for example) content. In general, conditions that result in a lower FFA content will lead to a higher loss of tocopherols. In most parts of the parameter zone, 50% loss of tocopherols will be expected. The deodorization study of randomized SL from fish oils and tricaprin indicated that SPD in comparison with batch deodorization gave a product of a poorer sensoric quality.

Different effects of diets rich in olive oil, rapeseed oil and sunflower-seed oil on postprandial lipid and lipoprotein concentrations and on lipoprotein oxidation susceptibility

Elevated concentrations of fasting and non-fasting triacylglycerol-rich lipoproteins (TRL) as well as oxidative changes of lipoproteins may increase the risk of ischaemic heart disease. To compare the effects of different diets rich in unsaturated fatty acids on the concentrations and in vitro oxidation of fasting and postprandial lipoproteins eighteen males consumed diets enriched with rapeseed oil (RO), olive oil (OO), or sunflower-seed oil (SO) in randomised order for periods of 3 weeks followed by a RO test meal. In the postprandial state the concentrations of cholesterol and triacylglycerol (TAG) in TRL were higher after consumption of OO compared with RO and SO (P<0.04), possibly related to differences in the fasting state. The propagation rates for VLDL and LDL oxidation were higher in the postprandial compared with the fasting state irrespective of diet. In the fasting state, the propagation rates were highest after SO (P<0.001), and in the postprandial state, SO gave rise to a shorter VLDL lag time (P=0.03) and a higher propagation rate than OO consumption (P=0.04). Overall, the SO diet resulted in a higher postprandial propagation rate of LDL (P<0.001) compared with RO and OO, while there was no effect of diet on LDL oxidation lag time. Our results suggest that RO and SO diets lower the postprandial cholesterol and TAG concentrations compared with OO, while RO and OO diets result in similar and lower in vitro susceptibility to oxidation of lipoproteins than SO.

Oxidative degradation and non-enzymatic browning due to the interaction between oxidised lipids and primary amine groups in different marine PL emulsions

Due to the beneficial health effects of marine phospholipids (PL) there is an increasing industrial interest in using them for nutritional applications including emulsified foods. This study was undertaken to investigate both oxidative and hydrolytic stability of marine PL emulsions in relation to the chemical composition of the marine PL used. Moreover, non-enzymatic browning reactions were also investigated. Emulsions were prepared by high pressure homogenizer using different concentrations and sources of marine PL. In some formulations, fish oil was added in order to study the effect of increasing levels of triglycerides in the emulsions. The oxidative and hydrolytic stability of emulsions was investigated through measurement of peroxide value, free fatty acids, and (31)P NMR during storage at 2°C for up to 32 days. The oxidative stability of marine PL emulsions during storage was further investigated through the measurement of secondary volatile compounds by solid-phase microextraction (SPME) and dynamic headspace (DHS) connected to gas chromatography (GC-MS). Non-enzymatic browning reactions were investigated through the measurement of Strecker derived volatiles, colour changes and pyrrole content. The results suggested that the oxidative stability of marine PL emulsions was significantly influenced by the chemical composition and the concentration of marine PL used to prepare them. Emulsions with good oxidative stability could be prepared from marine PL of high purity and high content of PL and antioxidant and low TAG content.

The choice of homogenisation equipment affects lipid oxidation in emulsions

Milk proteins are often used by the food industry because of their good emulsifying properties. In addition, they can also provide oxidative stability to foods. However, different milk proteins or protein components have been shown to differ in their antioxidative properties, and their localisation in emulsions has been shown to be affected by the emulsification conditions. The objective of this study was to investigate the influence of homogenisation equipment (microfluidizer vs. two-stage valve homogeniser) on lipid oxidation in 10% fish oil-in-water emulsions prepared with two different milk proteins. Emulsions were prepared at pH 7 with similar droplet sizes. Results showed that the oxidative stability of emulsions prepared with sodium caseinate was not influenced by the type of homogeniser used. In contrast, the type of homogenisation equipment significantly influenced lipid oxidation when whey protein was used as emulsifier, with the microfluidizer resulting in lower levels of oxidation.

Impact of primary amine group from aminophospholipids and amino acids on marine phospholipids stability: Non-enzymatic browning and lipid oxidation

The main objective of this study was to investigate the oxidative stability and non-enzymatic browning reactions of marine PL in the presence or in the absence of primary amine group from aminophospholipids and amino acids. Marine phospholipids liposomal dispersions were prepared from two authentic standards (phosphatidylcholine and phosphatidylethanolamine) and two purified PL from marine sources with and without addition of amino acids (leucine, methionine and lysine). Samples were incubated at 60°C for 0, 2, 4 and 6days. Non-enzymatic browning reactions were investigated through measurement of (i) Strecker derived volatiles, (ii) yellowness index (YI), (iii) hydrophobic and (iv) hydrophilic pyrroles content. The oxidative stability of the samples was assessed through measurement of secondary lipid derived volatile oxidation products. The result showed that the presence of PE and amino acids caused the formation of pyrroles, generated Strecker derived volatiles, decreased the YI development and lowered lipid oxidation. The lower degree of lipid oxidation in liposomal dispersions containing amino acids might be attributed to antioxidative properties of pyrroles or amino acids.

Effect of meal fat quality on oxidation resistance of postprandial VLDL and LDL particles and plasma triacylglycerol level.

This study was performed to examine the postprandial effects of meals containing dietary fats, with their natural fatty acid composition and tocopherol content, on the plasma triacylglycerols (TG) and tocopherols and on the resistance of VLDL and LDL to oxidation. On six separate days eighteen healthy male subjects were given low-fat meals (LF) or the LF meals enriched with sunflower oil (SO), rapeseed oil (RO), olive oil (OO), palm oil (PO), or butter (B) in a crossover design. The fat-rich meals all resulted in similar postprandial TG responses while the LF test meal did not increase plasma TG level. The postprandial plasma fatty acid profile changed to resemble the fatty acid composition of the ingested test fat. The alpha-tocopherol:gamma-tocopherol ratios in postprandial plasma and VLDL samples were greater than in the test fats. We found that the resistance of VLDL particles to oxidation in the postprandial state as assessed from lag time was increased after the PO-rich meal as compared with the SO-rich meal (p = 0.018), and the propagation rate was greater after the SO- and RO-rich meals compared with the others (p < 0.001). The resistance of LDL particles to oxidation was unaffected by the meals. In postprandial VLDL samples, the content of alpha-tocopherol was greater after the OO- and SO-rich meals compared with the meal rich in PO (P = 0.034 and 0.042 respectively). The gamma-tocopherol content of VLDL was highest after RO-meal as compared with all other test meals (P = 0.0019), and higher after SO as compared with B (P = 0.0148). Large individual differences were noted. In conclusion, meals enriched with different fats lead to the formation of VLDL particles with varying resistance to oxidation.

Stabilization of omega-3 oils and enriched foods using antioxidants

Abstract: Foods enriched with omega-3 polyunsaturated fatty acids (PUFA) are highly susceptible to lipid oxidation due to their high degree of unsaturation. Addition of antioxidants is therefore often necessary in order to prevent oxidation. However, antioxidant efficacy is not easy to predict in complex food systems as it is influenced by many different factors. This chapter will briefly discuss the major factors influencing antioxidant efficacy in heterophasic food systems enriched with omega-3 fatty acids. Moreover, the efficacy of different antioxidants in a range of different omega-3 enriched foods will be reviewed.

Oxidative Stability of Dispersions Prepared from Purified Marine Phospholipid and the Role of α-Tocopherol

The objective of this study was to investigate the oxidative stability of dispersions prepared from different levels of purified marine phospholipid (PL) obtained by acetone precipitation, with particular focus on the interaction between α-tocopherol and PL in dispersions. This also included the investigation of nonenzymatic browning in purified marine PL dispersions. Dispersions were prepared by high-pressure homogenizer. The oxidative and hydrolytic stabilities of dispersions were investigated by determination of hydroperoxides, secondary volatile oxidation products, and free fatty acids, respectively, during 32 days of storage at 2 °C. Nonenzymatic browning was investigated through measurement of Strecker aldehydes, color changes, and pyrrole content. Dispersions containing α-tocopherol or higher levels of purified marine PL showed a lower increment of volatiles after 32 days storage. The results suggested that tocopherol is an efficient antioxidant in PL dispersions or that the presence of α-tocopherol and pyrroles may be the main reason for the high oxidative stability of purified marine PL dispersions.

Lipolysis of different oils using crude enzyme isolate from the intestinal tract of rainbow trout, Oncorhynchus mykiss

Crude enzyme isolate was prepared from the intestine of rainbow trout. Positional specificity of the crude enzyme isolate was determined from both 1(3)- and 2-MAG products after in vitro lipolysis of radioactive-labeled triolein. The ratio of 2-MAG/1(3)-MAG was 2∶1, suggesting that the overall lipase specificity of the enzyme isolate from rainbow trout tended to be 1,3-specific; however, activity against the sn-2 position also was shown. In vitro lipolysis of four different unlabeled oils was performed with the crude enzyme isolate. The oils were: structured lipid [SL; containing the medium-chain FA (MCFA) 8∶0 in the sn-1,3 positions and long-chain FA (LCFA) in the sn-2 position], DAG oil (mainly 1,3-DAG), fish oil (FO), and triolein (TO). MCFA were rapidly hydrolyzed from the SL oil. LCFA including n−3 PUFA were, however, preserved in the sn-2 position and therefore found in higher amounts in 2-MAG of SL compared with 2-MAG of FO, DAG, and TO. Lipolysis of the DAG oil produced higher amounts of MAG than the TAG oils, and 1(3)-MAG mainly was observed after lipolysis of the DAG oil. The positional specificity determined and the results from the hydrolysis of the different oils suggest that n−3 very long-chain PUFA from structured oils may be used better by aquacultured fish than that from fish oils.