Influence of deep-fat frying process on phospholipid molecular species composition of Sardina pilchardus fillet
Introduction
Fish is an excellent source of nutrients such as essential amino acids, bioactive fatty acids, minerals, vitamins, chitin and antioxidants (Gebauer, Psota, Harris, & Kris-Etherton, 2006). Extensive epidemiological studies have demonstrated the protective role of fish and fish oil consumption against a wide range of diseases that are becoming more widespread in Western populations, including coronary heart diseases (Barringer and Harris, 2012, Calder and Yaqoob, 2009, Saravanan et al., 2010), type 2 diabetes (Malekshahi et al., 2012), inflammatory disorders (Simopoulos, 2008), cancer (MacLean, Newberry, Mojica, & Khanna, 2006) and neurodegenerative diseases (Freeman et al., 2006, Huang, 2010).
The potential health and nutritional benefits of fish consumption are mainly attributed to the lipid fraction, which is primarily composed of phospholipids (PLs) and triacylglycerols (TAG) exceptionally rich in ω3 polyunsaturated fatty acids (ω3 PUFA). The strong and positive biological activities of ω3 PUFA have been confirmed by extensive research over the past several decades and various governments and health organizations are currently recommending dietary intakes for total ω3 PUFA of 1.4–2.5 g/day (Lichtenstein et al., 2006, Harris et al., 2009). Nevertheless, in many societies of the Western style diet, the daily ω3 PUFA dose requirement is not always ensured.
Despite intensive investigation devoted to the achievement of an adequate dietary intake of ω3 PUFA, more recently, attention has also been focused towards the effects of different ω3 PUFA dietary forms (i.e., ethyl esters or PLs) (Neubronner et al., 2011). Within this context, fish PLs have created a great deal of interest (Burri et al., 2012, Dasgupta and Bhattacharyya, 2007, Shirouchi et al., 2007) since they have shown to be more efficient carriers of ω3 PUFA than fish TAGs in terms of ω3 PUFA absorption in different tissues (Parmentier, Al Sayed Mahmoud, Linder, & Fanni, 2007). This is particularly true when ω3 PUFA, such as eicosapentaenoic acid (EPA, 20:5ω3), docosapentaenoic acid (DPA, 22:5ω3) and docosahexaenoic acid (DHA, 22:6ω3), are mainly esterified at the sn-2 position, as observed in marine PLs. In addition, fish PLs have also exhibited anti-inflammatory and antitumor-promoting effects.
Sardine (Sardina pilchardus) is one of the most commercialized and consumed fishes in the Mediterranean area (Ismea, 2012, pp. 2–5). Sardine lipids have important nutritional characteristics because of their particularly high level of ω3 PUFAs, thus allowing the recommended dietary intake of PUFAs to be easily reached consuming less than three serving of this kind of fish per week and without the need for any supplementation (Bandarra et al., 1997, Cardenia et al., 2013, Pacetti, Balzano, et al., 2013, Pacetti, Mozzon, et al., 2013). However, most fish are consumed cooked, and considering that the culinary processes undoubtedly alter the content, the composition and the biological activity of the fish lipids, the nutritional value of the final cooked product is of major importance for human health (Nomikos, Karantonis, Skarvelis, Demopoulos & Zabetakis, 2006). Within the wide range of available cooking procedures, deep-fat frying is one of the most common food processing methods used for preparing a worldwide variety of foods, including fish. During frying both oil and food are modified, with ω3 long chain PUFAs in PLs representing the most heat-labile and oxidation-sensitive fatty acids. Over the past 30 years, several studies were undertaken to determine the effects of deep fat-frying and pan-frying on the fatty acids of fish species (Ansorena et al., 2010, Candela et al., 1998, Gladyshev et al., 2007, Sanchez-Muniz et al., 1992, Sebedio et al., 1993, Sioen et al., 2006, Weber et al., 2008, Zervou et al., 2012; Zhang et al., 2013). However, despite the increased interest in the nutritional and biological activities of ω3 PUFA rich PLs, there are no current data regarding the modifications of PLs profile that occur during the deep fat-frying procedure. Therefore, the aim of our work was to determine the effects on PL composition of edible muscle (fillet) of S. pilchardus of deep-fat frying performed using different culinary fats (extra virgin olive oil, conventional sunflower oil and high-oleic sunflower oil) and different temperatures (160 and 180 °C). The PL molecular species of the main fish PL classes (phosphatidylethanolamine, PE, phosphatidylcholine, PC) were determined by high pressure liquid chromatography (HPLC) coupled with a second order mass spectrometer (MS–MS) with electronebulization interface (ESI).
Beyond providing new information about the effect of the frying process on the ω3 PUFA rich PLs composition, this work helps to clarify the influence of different culinary fats and frying temperatures on the evolution of the PLs fraction, thus providing consumers and food industry with additional knowledge that can be used for the control and/or the preservation of the fried fish quality.
Section snippets
Material
Chloroform and methanol were HPLC grade from Lab-Scan (Dublin, Ireland); ammonia solution (30%) of analysis grade was from Carlo Erba (Milano, Italy). All other chemicals, with noted exceptions, were obtained from Sigma Chemicals Co. (St. Louis, MO). PLs standards including 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE, purity > 99%), 1,2-dipalmitoyl-sn-glycero-3-phosphocoline (DPPC, purity > 99%), 1-palmytoil-2-oleoyl-sn-glycero-3-phosphocoline (POPC, purity > 99%), 1,2-dipalmitoyl-sn
Separation of phospholipid classes and identification of phospholipid molecular species
The HPLC/ESI-MS-MS method previously developed for the characterization of phospholipid molecular species in fish and shellfish (Boselli, Pacetti, Lucci, & Frega, 2012) was applied to the qualitative and semi-quantitative analysis of the phospholipid molecular species in raw and fried sardine fillets. In the chromatographic conditions adopted, all PL classes eluted within 14 min in the following order: plasmalogen phosphatidylethanolamine (pPE) < PE < phosphatidylinositol
Discussion
The deep-fat frying process caused significant changes on PE and PC molecular species composition. Frying led to a significant relative increase of the proportion of the PE and PC species formed by the combination of palmitic and docosahexaenoic acids and to a slight (P > 0.05) depletion of the percentage of the PE and PC species formed by two docosahexaenoic acid residues in fried fillets. The PC composition resulted to be more affected by the frying process than PE. In fact, although the
Conclusions
Nowadays consumers demand for an improved quality of foodstuffs expecting that certain processed food would exhibit nutritional quality in addition to sensorial attributes as well.
Taking into account the nutritional relevance of food phospholipids, our finding contributes with original results concerning the effects of deep-fat frying on the fish phospholipid molecular species, needed to achieve the nutritional value of the final cooked fish. Noteworthy, most fish species, especially sardine,
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