Cocoa polyphenols ( Theobroma cacao ) as natural Amazonian antioxidant in sausage fresh.

Graphical Abstract Polyphenols ( Theobroma cacao ) are used as natural Epicatechin Abstract Ecuador is considered one of the richest countries regarding biodiversity and its Amazon region guests several flora and fauna species. Cacao ( Theobroma Cacao ) is a tropical fruit with high commercial and biological importance due to the presence of polyphenols with antioxidant activity. In the present work cocoa polyphenols ( Theobroma cacao ) are used as natural Amazonian antioxidant in sausage fresh. Experimental units of sausage samples with different percentages (0, 2, 4, 6%) of cocoa polyphenols in the formulation with an experimental size of 5 kg were made; once the sausage was prepared, samples of 100 g were taken to perform sensory analysis and 100 g of samples destined for microbiological analysis. The microbiological analysis was performed on each sample of the formulated product, analyzes on the newly elaborated samples (1 day of elaboration) and an analysis with samples was developed after an estimated time (30 days) to assess the differences and active action of the antioxidant as a natural Amazonian preservative were performed. 15 people evaluated sensory characteristics to the sausage samples through a tasting test. As main results the natural antioxidants use allows prolonging the product shelf life, which results in an increase in color stability, since it prevents the transition from myoglobin to metamyoglobin, as well as maintaining its organoleptic conditions unalterable, slowing down oxidative phenomena such as product rancidity or increasing resistance to bacterial growth, since antioxidants of polyphenolic nature have antimicrobial activity.


Introduction
For years, various strategies have been developed to prevent oxidative deterioration in products of meat origin through the use of antioxidants (Rostamzad et al., 2011). Most of these strategies have focused on limiting oxygen access to meat components susceptible to oxidation phenomena such as lipids and proteins. At the same time, new storage methods have been developed, such as vacuum packaging or packaging in a modified atmosphere in order to prevent the appearance of oxidation phenomena in the final product (Armenteros et al., 2012).
MOL2NET, 2017, 3, doi:10.3390/mol2net-03-xxxx 3 One way to reduce the occurrence of oxidation phenomena in meat and / or meat products is the use of antioxidants. The term antioxidant is generally attributed to any substance that is present at low concentrations, with respect to those of an oxidizable substrate and retards or prevents the oxidation of that substrate (Halliwell and Gutteridge, 1990). When antioxidant reacting with the free radical, it gives an electron oxidizing in turn and becoming a weak radical, with little or no toxic effects. In recent years it has been shown that a diet rich in plant polyphenols can improve health and decrease the incidence of cardiovascular diseases (Quiñones et al., 2012). In the present work cocoa polyphenols (Theobroma cacao) are used as natural Amazonian antioxidant in sausage fresh.

Location and duration of the experiment
The present research was carried out in the Agroindustry's Laboratory, located in the Amazon State University, Km. 2 1/2 via Puyo to Tena (Paso Lateral), province of Pastaza, between coordinates 0º 59 '-1 "S and at a length of 77º 49' 0" W, it is found in the Amazonian Region of Ecuador in the west of the province of Pastaza, at about 924 m.a.s.l. Temperature 18 to 24 ºC.

Experimental units
Experimental units were formed for each sample of chorizo with different percentages of Theobroma cacao polyphenols in the formulation (0, 2, 4, 6%) and an experimental size of 5 kg of prepared dough.
Once the sausage was prepared, samples of 100 g of each replicate sample were taken to perform sensory analysis and 100g of samples destined for microbiological analysis.

Microbiological analysis.
The microbiological analysis was performed on each sample of formulated product, analyzes were performed on the newly elaborated samples (1 day of elaboration) and an analysis with samples was developed after an estimated time (30 days) to assess the differences and active action of the antioxidant as a natural Amazonian preservative.

Sensory analysis and shelf life.
Through a tasting test, 15 people evaluated sensory sausage samples. An evaluation the product to know the shelf life in 30 days was made.

Sensorial analysis
40% of the evaluated peoples that correspond to 6 persons, likes sausage samples without any addition of natural antioxidant; 9 from 15 people like the product with 2% natural antioxidant addition, 7 people who represent 47% have similarity in the sausage with 4% natural antioxidant incorporated, finally, 6 people equivalent to 40% like it and they like the product with 6% antioxidants. It should be noted that

Microbiological analysis
The following table shows the significant reduction of microbial load in the stored sausages and that they contain polyphenols percentages incorporated in their formulation, while in the control product, since there is no protective agent, a microbiological growth can be observed (table 2). The use of natural antioxidants allows to prolong the useful life of the product which is in an increase of the stability of the color, since it avoids the transition from myoglobin to metamyoglobin, as well as maintains its MOL2NET, 2017, 3, doi:10.3390/mol2net-03-xxxx 5 organoleptic conditions unalterable slowing down oxidative phenomena as the rancidity of the product or increasing resistance to bacterial growth, since antioxidants of polyphenolic nature have antimicrobial activity (Naveena et al., 2008).
Consumers are becoming more aware of the toxicological implications of artificial additives in foods. The sun mushroom powder had no effect on microbiological stability. It is concluded that sun mushroom was effective in terms of the oxidative stability of pork sausage when added in powdered form at concentrations of 1.0%, 2.0% and 4.0%. Cocoa polyphenols (Theobroma cacao) presents good qualities as an antioxidant in the use of sausages, allowing a better conservation and providing organoleptically unique qualities. Chorizo with 2% presents the best sensory characteristics and most welcome in the respondents. Meat products made with the addition of natural antioxidants from the Amazon provide, in addition to their basic nutritional properties, providing consumers with food that allows them to obtain better quality, health and life expectancy.