Meat Science Carbonyl Beef Method Colorimetric Oxidation

• Periodical List
• Ital J Nutrient Saf
• v.vii(3); 2018 Sep 26
• PMC6240832

Ital J Food Saf. 2018 Sep 26; 7(iii): 7342.

Olive mill wastewater phenolic concentrate equally natural antioxidant confronting lipid-protein oxidative deterioration in chicken meat during storage

Rossana Roila

ⁱDipartimento di Medicina Veterinaria, Università degli Studi di Perugia

Andrea Valiani

²Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche "Togo Rosati", Perugia

Dino Miraglia

¹Dipartimento di Medicina Veterinaria, Università degli Studi di Perugia

David Ranucci

ⁱDipartimento di Medicina Veterinaria, Università degli Studi di Perugia

Claudio Forte

²Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche "Togo Rosati", Perugia

Massimo Trabalza-Marinucci

ⁱDipartimento di Medicina Veterinaria, Università degli Studi di Perugia

Maurizio Servili

ⁱⁱⁱDipartimento di Scienze Agrarie, Alimentari ed Ambientali, Università degli Studi di Perugia

Michela Codini

⁴Dipartimento di Scienze Farmaceutiche, Università degli Studi di Perugia, Italy

Raffaella Branciari

^oneDipartimento di Medicina Veterinaria, Università degli Studi di Perugia

Received 2018 February 13; Revised 2018 Jun 12; Accepted 2018 Jul 16.

Abstract

Considering that many plant-derived substances show antioxidant and antimicrobial properties, natural antioxidant administered through feed in livestock animals could increment the shelf life of meat and meat products. The aim of this work was to study the effect of olive oil by-products on chicken meat lipid and protein oxidation and oxidative stability during storage. Two hundred and ninety-seven 22-day-one-time fast growing (Ross 308) female chicks were randomly assigned to iii experimental grower-finisher diets: i) a basal control diet (CTR), two) CTR diet supplemented with a low dosage (4.viii%) of olive manufactory wastewater extract (Fifty-OW) and iii) CTR diet supplemented with a high dosage (9.ix%) of olive manufacturing plant wastewater extract (H-OW). Breast meat of animals belonging to each experimental group was sliced, overwrapped with oxygen-permeable packaging and analysed at three different storage times (zero, iii and seven days). At the three sampling times considered, for all samples, colour coordinates (a*), saturation index, Hue angle, peroxide value, thiobarbituric reactive substance, carbonyl assay and the oxygen radical absorbance capacity determinations were performed. No differences in color were detected amongst the groups in all the sampling times considered. In decision, the supplementation of chicken diet with olive mill wastewater extract (OW) affected oxidation of meat, retarding lipid and protein oxidation and improving antioxidant activity during storage.

Fundamental words: Poultry, TBARS, Carbonyls, Oxidative Stability

Introduction

Oxidation is ane of the major causes of quality deterioration limiting meat acceptability. Oxidative deterioration, in whatever blazon of meat, is responsible for discoloration, development of off-flavour, formation of toxic compounds and brusk shelf-life. Meat becomes susceptible to oxidative deterioration due to high concentrations of unsaturated lipids, heme pigments in specific species, metallic catalysts and a range of oxidizing agents in the muscle tissue (Branciari et al., 2015a).

Oxidation in lipid and protein fractions of meat has been demonstrated as the main non-microbial cause of quality deterioration during processing. This happens because lipids and proteins in meat are easily susceptible to oxidative damages due to the rapid depletion of endogenous antioxidants after slaughter (Xiao et al., 2013). However, the susceptibility of meat to oxidation has also been found to be influenced by animate being breed and specie, muscle type and anatomical location (Min et al., 2008). Moreover, the type of diet consumed by animals during the last office of their life has a great influence on the susceptibility of meat to oxidation.

In contempo years special attention has been paid to several natural antioxidants (Insani et al., 2008; Ranucci et al., 2013; Roila et al., 2016) that tin exist applied straight in food every bit technological strategies or through fauna diet to reduce or prevent oxidative procedure in muscle food (Dal Bosco et al., 2012; Novelli et al., 2014). Technological strategies involve the application of antioxidants straight into the meat and meat products or by blanket packaging materials with plant extracts to improve the oxidative stability of the products. In dietary manipulations antioxidants are introduced into the muscle via feed.

Olive oil by-products are amongst those plant derived natural compounds that can exist used as potential sources of antioxidants for muscle food preservation and nutritional quality improvement (Branciari et al., 2017) because of their loftier content of phenolic substances (Novelli et al., 2014). In particular, olive factory wastewater is characterised by a high content in hydroxytyrosol (3,4-DHPEA), tyrosol (p- HPEA), secoiridoids derivatives, in detail the dialdehydic course of decarboxymethyl elenolic acid linked to iii,4- DHPEA or p-HPEA (3,4-DHPEA-EDA or p-HPEA-EDA) and verbascoside (Branciari et al., 2017). No information are currently available on the furnishings of dietary supplementation with olive manufacturing plant wastewater on chicken meat oxidation during storage.

The aim of this piece of work was to evaluate the event of dietary handling with olive mill wastewater on lipid and protein oxidative stability during storage.

Materials and Methods

Two hundred ninety-7 22-days-old female Ross 308 chicks, previously fed with the aforementioned starter diet from twenty-four hour period i to day 21, were allocated for a total of xx days in an experimental farm (Umbria, Italia) under environmental condition simulating those nowadays in conventional intensive systems. The subjects were randomly assigned to one of the iii experimental dietary groups and so divided in three replicates of 33 birds each (experimental units). Groups were fed three different diets: a basal grower-finisher control diet (CTR); CTR feed added with low dosage (4.8%) of olive factory wastewater (50-OW); CTR feed added with high dosage (ix.9%) of olive manufacturing plant wastewater (H-OW). The diets resulted isoenergetic and isonitrogenous and were formulated to meet National Research Quango standards for broiler chickens (NRC, 1994). Olive mill waste water polyphenol content of the dissimilar feeds was analyzed resulting 263.2 mg/kg for the L-OW diet and 556.five mg/kg for the H-OW feed. Olive manufacturing plant wastewater supplement was obtained as reported by Branciari et al. (2016) from processing of Italian cultivar Moraiolo of Olea europea using a filtration system based on progressive permeability membranes.

At the end of the trial, at 41 days of historic period, the animals were transported and slaughtered at a local slaughter-house. Immediately after slaughter the carcasses were promptly chilled to four°C. After 24 h the Pectoralis major was removed from carcasses, sliced, randomly assigned to retail packs overwrapped with oxygen-permeable packaging and stored at iv±1°C for 7 days. Packed meat samples were analysed during storage for colour, peroxide value (POV), thiobarbituric reactive substances (TBARS), carbonil assay and ORAC^FL.

Lipid oxidation

For the evaluation of primary meat lipid oxidation, the POV was adamant, during storage, as reported past Branciari et al. (2015b). Secondary meat lipid oxidation was assessed using the TBARS according to Tarladgis et al. (1960); the values were expressed equally mg malondialdehyde (MDA)/kg.

Poly peptide oxidation

Protein oxidation, measured by total carbonyl content, was evaluated by derivatization with DNPH co-ordinate to the method described by Reznick and Packer (1994) with slight modifications: ii g of meat were thawed, minced and and so homogenized 1:ten (w/5) in phosphate buffer (pH 7.iv), consisting of 50 mM NaH²PO^four and 1mM EDTA, using an ultraturrax homogenizer for 30 southward. The homogenates were divided in ii equal aliquots of 0.1 mL. Afterwards proteins were precipitated in both aliquots by adding 1 mL of ten% TCA and centrifuged for five min at 5000 rpm. Finally, the supernatants were removed, one pellet was treated with 1 mL 2 N HCl (for quantifying poly peptide concentration) and the other one with an equal volume of 0.2% (westward/5) DNPH in 2 N HCl (for carbonyl concentration measurement). Both samples were incubated for 1 h at room temperature (shaken every 15 min). Afterwards the samples were precipitated with 1 mL of x% TCA and washed twice with 1 mL of 1:ane ethanol/ethyl acetate (v/five), shaken and centrifuged for five min at 10000 rpm. The pellets were and then dissolved in 1.v mL of twenty mM sodium phosphate buffer (pH 6.5) containing vi Thousand guanidine hydrochloride, stirred and centrifuged for 2 min at 5000 rpm to remove insoluble fragments. Protein concentration was calculated from absorption at 280 nm using bovine serum albumin as standard.

The number of carbonyls was measured at 370 nm and expressed as nmol of carbonyl per mg of poly peptide using the adsorption coefficient for the protein hydrazones (21.0 mM ^-1 cm^-one).

Color measurement

The color measurement was performed, during each of the sampling intervals, subsequently a 30 minutes flower period at refrigeration temperature using a CR400 Minolta Chromameter (Minolta, Osaka, Japan – low-cal source of D65 calibrated against a standard white tile). The results were expressed as redness (a*), hue value (tan-1 b*/a*) and saturation alphabetize, or chroma ((a*two + b*ii)one/two), (CIE L*a*b* colour system, 1976).

Antioxidant capacity of meat

The antioxidant capacity of meat (x samples for each treatment in triplicate) was adamant using the oxygen radical absorbance capacity method (ORAC^FL) based on the fluorescence decay rate of a probe in the presence of a radical oxygen species (ROO) compared with that of a reference standard, Trolox (6-hydroxy- 2,5,7,8-tetramethylchroman- ii-carboxylic acid, Sigma-Aldrich, Steinheim, Germany). The extraction was performed on ii g of meat sample co-ordinate to Branciari et al. (2015b).

The ORAC^FL assays were carried out on a FLUOstar OPTIMA microplate fluorescence reader (BMG LABTECH, Offenburg, Germany) at an excitation wavelength of 485 nm and an emission wavelength of 520 nm. The procedure was based on the method of Branciari et al. (2015a). Briefly 2,20-azobis (ii- methylpropionamide) dihydrochloride (AAPH; Sigma-Aldrich) was used as a peroxyl radical generator, Trolox was used equally a reference antioxidant standard, and fluorescein was used as fluorescent probe. A 100 μL book of diluted sample, bare or Trolox scale solution (x-fourscore μmol) was mixed with 1 mL of fluorescein (80 nM), then 200 μL of each mixture was placed in a well of the microplate. The microplate was placed in the reader and preincubated for 15 min at 37°C. To each well, lx μL of AAPH was automatically added to initiate the reaction. The fluorescence (FL) was measured every 1.9 min. All the reaction mixtures were prepared in duplicate, and at to the lowest degree 3 independent assays were performed for each sample. The last ORAC^FL values were calculated past using a linear regression equation (Y = a + bX) to describe the relationship between the Trolox concentration (Y) and the net area under the FL decay bend (X). Linear regression was used in the range of 10-80 lM Trolox. The data are expressed as micromoles of Trolox equivalents (TE) per gram of sample (μmol TE 1000 ^-i) by applying the following formula:

where Ctrolox is the concentration of Trolox, yard is the sample dilution factor, and AUC is the expanse beneath the fluorescence decay curve of the sample, the blank and Trolox, respectively, calculated past applying the post-obit formula (Ou et al., 2001) in a Microsoft Excel spreadsheet (Microsoft, Washington, DC, Usa)

where f1 is the initial fluorescence reading at t = 0 min and fi is the fluorescence reading at fourth dimension i. The net AUC for each sample was obtained by subtracting the AUC of the respective blank from that of the sample.

Statistical analysis

The information were analysed using the GLM procedure of SAS (2001). ANOVA model was used with diet (CTR, L-OW and H-OW) and time (twenty-four hour period 0, mean solar day 3 and day seven) as fixed variables. The differences of the means were analysed using Tukey test and were considered to exist significant when P<0.05.

Results

The POV, TBARS values and the amount of carbonyl compounds are reported in Table i. POVs showed an increase at 3 days of storage than decreased at 7 days. The POV's value was college in CTR sample at 0 and three days of storage. At seven days of storage higher value of POV was registered for Depression and H-OW than CTR sample, considering lipid peroxides were cleaved to yield secondary oxidation products. The analysis of TBARS showed an increment during storage. Higher values for CTR sample than Fifty-OW and H-OW were recorded at 0 and 3 and 7 days of storage.

Table i.

Outcome of storage fourth dimension on peroxide value (POV meq O²kg^-i fat), thiobarbituric acid-reactive substances (TBARS mg MDA kg^{- i}) and carbonyls (nmol/mg protein) in breast meat stored at iv°C

Days of storage	Sample	POV	TBARS	Carbonyls
0	CTR	1.39bX	0.56bX	one.77bX
	L-OW	0.89aX	0.38abX	1.35aX
	H-OW	0.77aX	0.31aX	ane.23aX
	CTR	2.35by	0.88bY	i.93bY
	Fifty-OW	1.63aY	0.48aY	1.49aX
	H-OW	ane.57aY	0.41aY	1.33aY
7	CTR	0.31bZ	1.lxbZ	2.fifteenpast
	L-OW	0.53aZ	0.76aZ	1.60aX
	H-OW	0.61aX	0.69aZ	1.55aY
SEM	-	0.09	0.132	0.078
P-value	T	<0.001	<0.05	<0.001
	D	<0.001	<0.05	<0.001
	TXD	<0.001	0.057	0.897

For the colour parameters considered no deviation during storage, neither amidst samples, were detected for redness (a*), Hue angle and saturation index. The effect of fourth dimension and diet was evident for all lipid and poly peptide oxidation parameters but no for the color (Figures i-three).

The results obtained for the ORAC_FL determinations in meat samples are reported in Effigy four. Differences were recorded for CTR versus Fifty-OW and H-OW groups showing higher values in the antioxidant activeness compared to control in all fourth dimension of storage.

Effect of storage time on ORAC_FL of breast meat stored at 4°C.

Discussions

The results of the nowadays study testify that supplementing craven nutrition with phenolic compounds trough olive oil byproducts protects meat against lipid and protein oxidation. Chemical deterioration, and in particular lipid oxidation, is 1 of the main factors able to limit the shelf-life of meat. The inclusion of natural antioxidants in creature diets has been reported past various authors as a fashion to slow down meat lipid oxidation during storage, therefore improving the quality and the conservation of products (Ranucci et al., 2013; Luciano et al., 2013).

The POV value increased and thereafter decreased with the time of storage, moreover the decomposition of hydroperoxides into secondary products appears to increment at a higher rate in the CTR compared with the treated samples. These data are in accordance with those of Hwang et al. (2013), who reported that POV values of cooked pork patties added with natural polyphenols, increased and thereafter decreased with the fourth dimension of storage and highlighted that the decomposition of hydroperoxides into secondary products resulted delayed in treated samples.

Meat from animals fed dietary OW supplementation showed lower TBARS values during storage when compared to the control grouping. These results agree with other authors who establish a reduction of lipid oxidation measured trough TBARS during storage in lamb and beef muscles after olive oil past-products dietary supplementation (Luciano et al., 2013; Branciari et al., 2015a). Tufarelli et al. (2016) constitute an improved antioxidant defence organisation and a reduced TBARS level in chicken liver following dietary supplementation with extra virgin olive oil. Other authors have reported the beneficial furnishings of olive polyphenols on the oxidative status of meat measured through TBARS (Dal Bosco et al., 2012; Branciari et al., 2017), demonstrating their antioxidant effect in meat of animals fed olive phenolic compounds.

The extent of poly peptide oxidation was evaluated by the germination of poly peptide carbonyls. There are many reports on the biochemical changes in different meats during storage, about of which have focused on lipid oxidation (Abdel-Kader, 1996; Pikul et al., 1984). Lilliputian piece of work has been done on protein oxidation of meat obtained from fauna fed natural antioxidant. Zhang et al. (2011) reported an increase in lipid and poly peptide oxidation in the breast muscles of birds that had been fed an oxidized oil diet compared to antioxidant-supplemented and command diets. Jongberg et al. (2011) demonstrated the positive issue of grape extract on protein oxidation in arctic stored beef patties. A significant subtract in carbonyl formation in meat added 3% (due west/w) constitute fruit extracts compared to control burger patties was observed in the study past Ganhao et al. (2010). Protein oxidation has been shown to induce a number of changes in proteins, such as modification of amino acid side chains, germination of poly peptide polymers, loss of solubility, increment in carbonyl groups, change in amino acid composition and increase in proteolytic susceptibility (Levine et al., 1990; Xiong, 2000). This modification of muscle proteins, because of denaturation and proteolysis, induces changes in meat quality including texture traits, colour, aroma, flavor and water-holding chapters.

Considered the foregoing it is articulate that lipid and poly peptide oxidations are closely associated with deteriorative processes able to affect the overall quality of meat and meat products.

Meat color was non affected by the phenolic compounds, opposite to what was observed in meat of other species (Branciari et al., 2015a). This result might be due to the characteristics of P. major muscle which is mainly composed past α-white fibres that contain a lower amount of iron compared to the α-carmine or β-red blazon fibres and for this reason are less susceptible to color modification (Branciari et al., 2015b).

The results obtained from ORAC_FL assay showed the increased antioxidant activity of treated groups compared to command.

The effect of dietary OW on meat antioxidant activity might be related to several factors, including the presence of bioactive molecules in tissues (muscle and liver) of animal fed with olive oil polyphenols as reported past Branciari et al. (2017). Indeed, in that location is prove that polyphenols supplementation can exert a protective event on α-tocopherols, a defensive barrier against lipid oxidation from oxidative disuse (Bars-Cortina et al., 2018). In addition, polyphenols were shown to affect the expression of genes involved in lambs' stress response and in primary homeostasis pathways, in both liver and muscle, increasing the expression of MYLK genes and improving the meat quality in female lambs (Sabino et al., 2018).

Conclusions

The results of the nowadays experiment demonstrated that olive polyphenols used in chicken diets could delay meat lipid and protein oxidation without affecting its color stability. Further studies are needed in social club to better clarify the role of olive mill wastewater bioactive compounds in the shelf-life of chicken meat, packaged with different systems. Moreover, additional data is required to define the most suitable concentration of polyphenol compounds to be used in animal diets able to exert at best their activity.

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Consequence of storage time on CIE a* value of breast meat stored at 4°C.

Effect of storage time on Hue angle of breast meat muscle stored at four°C.

Result of storage time on Saturation index of breast meat stored at four°C.

Funding Argument

Funding: this work is part of a research project funded by the Italian Ministry of Health (RC 007/2012 "Valutazione dell'effetto di una dieta integrata con polifenoli derivanti da reflui dell'industria elaiotecnica sulle caratteristiche igienico-sanitarie, chimico-fisiche, reologiche e organolettiche della carne fresca e delle preparazioni di carne: filiera del pollo da carne").

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Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6240832/

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