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Yildiz-Turp G, Sedaroglu M. Beside the traditionally used antioxidants in meat and fish also a wide variation of herbs, spices and fruits are used more and more as additives with antioxidative capacity [ 13 - 17 ].
In the recent years a lot of research has been carried out evaluating these natural substances as antioxidative additives in food products leading to novel combinations of antioxidants and the development of novel food products [ 17 - 20 ].
The high antioxidant capacity of these plant parts is particularly due to their content of different phenols, anthocyanins and ascorbic acid, which can act as radical scavengers [ 21 ]. In addition to their antioxidative capacity, many of this natural substances have positive effects in the human body and documented health benefits and are therefore highly appreciated food additives [ 22 - 27 ].
So a combination of foods rich in omega 3 PUFA and plant substances rich in phenols and anthocyanins might result in nutritionally very valuable novel food products. These products could play and important role in the prevention of specific chronic-health problems beside dietary supplements where PUFA, probiotics and super-fruits are achieving particular interest in the recent time [ 23 , 28 ].
Finally nutritionally dense meals may be of interest and importance for people with particularly high nutritional demands, e. For animal foods there are always two possible ways to include antioxidants: Via the feed or post mortem during the processing. Depending on the type of antioxidant, the one or the other way will be more effective. In general fat soluble antioxidants like tocopherol are more effective when present in the feed, while water soluble ones like vitamin C are more effective when added during processing [ 30 , 31 ].
In addition there are synergistic effects between different antioxidants as for example shown for tocopherol and ascorbic acid [ 32 ] so a good combination of all available tools might be able to boost antioxidative protection for certain products. The present chapter will give an overview of the main used and tested antioxidants, synergistic effects and the possible increased nutritional value. Feeding effects as well as a variation of processing and preserving methods for animal products from both very traditional and most recent techniques will be presented and their influence on oxidative stability will be elucidated.
Lipid oxidation is omnipresent in meat and fish and their products. Especially in products with a high amount of unsaturated FA, oxidation leads to rancidity, off-flavour and taste and to the formulation of toxic substances [ 2 , 33 , 34 ].
In the food industry a great deal of research and attention is spend on the on-going oxidative processes. The main aim is always to protect the raw material and the products as good as possible from oxidation through the whole process and during storage. In order to get a whole picture about lipid oxidation, it is important to know some basics about lipids and FA. The C atoms in the chain can either be saturated or unsaturated meaning they form double bonds between each other.
The FA are generally named in the scheme X:Y n-z where X is the number of carbon atoms in the chain, Y the number of double bonds and z the number of the last carbon atom with a double bond counted from the methyl end see Fig. Linolenic acid, n The n stands in spoken language for omega so a FA with the last double bond at the third carbon atom from the methyl end is an omega 3 FA while the one with the last double bond at the sixth carbon atom from the methyl group is an omega 6 FA and so on.
A very good in depth review about the classification and chemistry of FA and also about their biological functions has been done by [ 35 , 36 ].
The reactivity of unsaturated FA increases with their chain length and number of double bounds [ 37 , 38 ]. Beside the number of double bonds also the placing of the double bonds and the form of the FA determine their oxidative reactivity. In general the n-3 FA are more prone to oxidation than the n-6 and those are more prone to oxidation than the n-9 FA [ 38 ].
In animal tissues the lipids are usually divided into two main classes: polar lipids PL and neutral lipids NL. NL consist mainly of triacylglycerols TAG which are three FA bound to a glycerol molecule, and minor amounts of mono- and diacylglycerols, whereas PL include mainly phospholipids which are diacylglycerols including a phosphatic acid derivate [ 39 ]. TAG serve mainly as an energy source, whereas phospholipids are mainly constituents of the cell and organelle membranes being essential for their functionality and fluidity [ 39 - 41 ].
Phospholipids are in general more unsaturated due to their functionality and therefore also more prone to oxidation. In addition free FA FFA can occur in raw or processed tissues due to enzymatic breakdown of acylglycerols or phospholipids. The complicated thing about oxidation is that once it started a cascade of reactions will occur with each new molecule increasing the reaction speed and variability Fig.
The kinetics of oxidation in meat and meat products are described by [ 38 ] and [ 42 ]. Oxidation leads to the formation of lipid radicals L.
Auto oxidation in meat and fish can be initiated by light, heat, presence of metal ions and radicals. Very low concentrations of radicals are needed to start the reaction. Once initiated, oxidation propagates in a chain reaction steps In the termination reactions, lipid peroxides LOO. In meat and muscle there are different possibilities to measure the degree of oxidation. The most used ones are listed very briefly here to facilitate the understanding of oxidation parameters used in this chapter:.
The peroxide value: determines the amount of hydroperoxides, which are among the primary products. However, as the peroxides are not stable and react further the results have to be evaluated carefully as, with on-going oxidation the peroxides first increase and reach a maximum but after a while the reaction speed towards secondary oxidation products is faster and the peroxide value decreases again [ 43 ]. Thiobarbituricacid TBA reacts with malondialdehyd a secondary oxidation product from PUFA with 3 or more double bonds to a pink complex that can be measured at nm.
However the problem with that method is, that other substances also form coloured complexes with TBA and might result in wrong estimation of the oxidation status [ 44 ]. Iodine value: A very traditional method which is still used sometimes to measure the iodine value as a number for the amount of lipid double bonds and the decrease of that number over time as a sign for oxidation.
Volatile lipid oxidation products by Headspace GC-MS: During the last decade also more advanced methods have been used more and more for evaluation of oxidation. Content of Hexanal and other volatiles has been shown to give a quite good picture of oxidation status and mechanisms [ 45 , 46 ]. However as these measurement are quite time consuming and expensive they are still not used routinely.
But as the FFA are oxidised faster than bound FA, they can be regarded as a measurement for increased oxidative reactivity of the muscle or product. Hypothetical autoxidation of a polyunsaturated lipid as a function of time [ 47 ]. Antioxidants can be introduced into the muscle by different means. Coming first in the natural chain from farm to fork would be to add the antioxidants via the feed.
Also in the feed antioxidants are needed, to stabilize the lipids in the feed during storage, especially true is that for fish feed with high contents of PUFA. The main used antioxidant in feeds is the fat soluble Vitamin E, normally added in the form of tocopherol acetate.
These include the tocopherols with a saturated phytyl side-chain, Fig. The water soluble vitamin C, ascorbic acid is another antioxidant used in feeds Fig. However studies have shown that in the live animal tocopherol shows a greater effect, while ascorbic acid works better added post mortem [ 30 , 31 ]. Structure of ascorbic acid.
A third group of natural occurring antioxidants are the also fat soluble carotenoids, the precursors of retinol vitamin A. They are for example found in corn. Carotenoids are hydrocarbons built from eight isoprene bodies 40 C atoms Fig. Due to their structure and the conjugated double bonds, both vitamin E and the carotenoids, are radical scavengers that can build relatively stable radicals. In addition, carotenoids, tocopherols and tocotrienols are quenchers for singlet oxygen.
Carnosine Fig. For example [ 30 ] suggested using a combination of feed additives and post mortem added antioxidants as for example a feed supplementation with a-tocopheryl acetate and post mortem applied carnosine. Structure of carnosine. Squalene is triterpene 30 C atoms Fig. It has similar to the carotenoids conjugated double bonds and can hence build stable radicals and has been investigated as possible antioxidant [ 50 ].
Significant amounts of squalene in plant sources are detected in e. Structure of squalene. As mentioned before, there is also a growing interest to use novel sources of natural antioxidants for feeds, as for example from diverse vegetables [ 53 , 54 ] or spices [ 55 ] or from more exotic sources as algae and lichen [ 56 , 57 ].
Moreover, there are also always interactions between different nutrients [ 59 - 61 ], which have to be taken into consideration when planning how to achieve antioxidative protection of animal foods. For example did high dietary lipid increase also muscle astaxanthin accumulation in salmon Salmo salar [ 60 ].
Astaxanthin is a carotenoid that gives the pink colour to salmon muscle but can also act as an antioxidant. However, if this mechanism is also valid for fish and other mammals, remains to be investigated. Concerning the oxidation occurring in the feeds during storage [ 62 ] showed that ascorbic acid could protect vitamin E from oxidation in the diet for hybrid tilapia.
Also for the nutritional status of the animals the dietary added antioxidants are of importance. Low dietary vitamin C content has shown to increase requirement of vitamin E in juvenile salmon [ 64 ], suggesting that the deficiency of one antioxidant will lead to the increased use of the available ones. However, bioavailability, efficiency and interactions with other substances might vary between different species as summarized by [ 63 ].
Oils and animal foods always contain a small amount of metals which are too difficult to remove, as for example iron from myoglobin, hemoglobin and the iron storage protein ferritin, or copper, zinc and heavy metals that are present in enzymes and metalloproteins [ 40 , 65 ]. Another source of metals in animal food products are the machines used during processing, from which minor amounts of iron can get into the products either by abrasion or due to acidic dissolving of metals from the surface.
A third source can be migration of metals from the packaging. These metals are present in so low amounts that they do not have a physiological effect; however, they can have pro oxidative effects [ 66 ]. Salt is used for the preservation of meat and fish. Due to its water activity lowering effect and the withdrawl of free water, salt decreases the solubility of oxygen as well as the activity of enzymes and bacteria. In addition chloride ions are also toxic to certain microorganisms.
However it is also a pro-oxidant [ 67 ]. The more and longer a product is exposed to light and oxygen, the higher is the risk and speed of oxidation.
When fish or meat is cut into pieces or minced, the surface is substantially increased and thereby the accessibility for oxygen.
As light and increased temperature enhance oxidation [ 40 , 68 ], during processing temperature and the processing time should be kept as low and short as possible respectively. Various processes including cooling, salting, drying, smoking and heating have been used for a long time to preserve meat and fish and to obtain a variety of products with characteristic organoleptic characteristic [ 2 , 69 , 70 ] Processing is a primarily way to preserve meat, but also adds to its value.
However, different processing steps can also negatively affect meat quality, and change for example lipid quality traits. Heating of meat and meat products e. Use of antioxidants during processing or alternative more gentle processing methods can reduce these negative effects.
Preservation of meat quality is an important criterion for its shelf life, since raw, chilled meat has traditionally been a perishable product [ 1 , 72 ].
In order to prolong the chilled storage time advanced packing techniques or various additives are used in addition, which will be described in more detail in the following. To make these ice slurries even more effective different additives to fish as well as to the ice slurry have been used. As a result, the quality of meat and meat products deteriorate when fat oxidizes and develops off-flavors. Lipid oxidation increases the conversion rate of oxymyoglobin bright red color to metmyoglobin brown discoloration and subsequently impacts the physical appearance of meat and meat products [1].
Meat is also very susceptible to spoilage and microbial growth during slaughtering and post-slaughter handling. Therefore, meat suppliers use various food additives to extend the shelf life of meat and meat products.
Nevertheless, due to increasing demands for clean label solutions, extensive work has been conducted to identify novel and natural extracts with potential applications in meat and meat products. Studies using natural extracts with potential applications in meat and meat products were reviewed by Kumar and colleagues. Examples of the natural extracts, active ingredients, and applications that have been studied can be seen in Table 1.
TABLE 1. Antioxidants from natural sources with the potential applications in meat and meat products. The most commonly used naturally sourced antioxidants are phenolic compounds such as phenolic acids, tocopherol, and flavonoids. Phenolics prevent lipid oxidation through different mechanisms—by functioning either as free radical scavengers, metal chelators, or singlet oxygen quenchers.
The antioxidative potential of phenolics depends on their skeleton structure. The number and location of functional groups, such as free hydroxyl OH groups, is just one example. For instance, phenolics with a higher number of OH groups and ortho-3,4-dihydroxy structures will have higher antioxidative properties [3].
In addition, some phenolics, such as carnosic acid from rosemary, and catechin from green tea, not only have OH groups that can donate hydrogen to free radicals, but also contain vicinal -OH groups that can chelate metals. Consequently, combinations of natural extracts can potentially deliver synergistic effects and improve antioxidant performance in preventing lipid oxidation.
Such combinations can also reduce the effective dosage of each extract, thus minimizing impacts on flavor and color. Fresh meat, including sausages, are a major category of meat retail sales, both in the United States and Europe. Fresh sausages contain a high level of fat and are prepared from fresh comminuted meat from different meat types such as pork, chicken, beef, and so on.
Consequently, synthetic antioxidants such as BHA, BHT, and propyl gallate are commonly used to extend the shelf life of fresh sausages in the US market. Three main mechanisms responsible for the spoilage of sausages are lipid oxidation, microbial growth, and enzymatic autolysis such as proteolysis and lipolysis. Of these, microbial growth is the main cause of spoilage for sausages [4, 5].
For this reason, naturally sourced ingredients comprising antioxidative and antimicrobial agents were developed in our laboratory to delay the spoilage of pork sausages from different mechanisms, and to meet the different needs of US, UK, and EU markets. A storage study was conducted to investigate the efficacy of a natural blend comprising rosemary extract and buffered vinegar to extend the shelf life of British pork sausages.
The obtained results showed that SMBS is only effective in stabilizing the color of pork sausages for the first few days, whereas naturally sourced ingredients were superior in maintaining the color of pork sausages even after 14 days of storage Fig. In addition, these naturally sourced ingredients provided a performance comparable to that of SMBS in inhibiting the growth of spoilage bacteria total plate count of 3.
The degree of lipid oxidation in sausages was monitored through the hexanal level, and using other selected oxidation markers such as heptanal, 1-octenol, and nonanal Fig.
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