Enhancing the quality and lipid stability of chicken nuggets using natural antioxidants
© The Author(s). 2017
Received: 16 January 2017
Accepted: 24 May 2017
Published: 8 June 2017
Current day consumers prefer natural antioxidants to synthetic antioxidants because they are more active. However, the activity generally depends on the specific condition and composition of food. The aim of this study was to investigate the effect of wheat germ oil and α-lipoic acid on the quality characteristics, antioxidant status, fatty acid profile, and sensory attributes of chicken nuggets.
Six types of diets were prepared for feeding the chickens to evaluate the quality of nuggets made from the leg meat of these experimental animals. These included control, diet enriched with wheat germ oil (WGO), which is a rich natural source of α-tocopherol (AT), diet with added AT or α-lipoic acid (ALA), diet with a combination of either ALA and WGO (ALA + WGO) or ALA and synthetic AT (ALA + AT). ALA has great synergism with synthetic as well as natural AT (WGO).
The diet with WGO and ALA showed the best potential with respect to both antioxidant activity and total phenolic content. HPLC results revealed that the chicken nuggets made from WGO + ALA group showed maximum deposition of AT and ALA. The stability of the nuggets from control group was found to be significantly lower than that of nuggets from the WGO + ALA group. Total fatty acid content too was higher in the nuggets from this group. The poly unsaturated fatty acids (PUFA) were found to be higher in the nuggets from the groups fed with a combination of natural and synthetic antioxidants.
It is concluded that the combination of natural and synthetic antioxidants in the animal feed exerts a synergistic effect in enhancing the stability and quality of chicken nuggets.
Using synthetic or natural antioxidants is a major strategy used to prevent lipid oxidation. The current trend in the food industry is using natural additives in preference to synthetic additives . The growth of the animals and the antioxidant potential of the meat from these animals can be improved by the addition of antioxidants as dietary supplements to animal feed. The addition of natural antioxidants to animal feed enhances the lipid stability, improves the sensory attributes of meat, and improves the functionality of meat products [2, 3].
The quality of meat increased by the addition of antioxidants in the animal feed . Different types of antioxidants are added to the animal feeds for different purposes. The use of synthetic and natural antioxidants has synergistic effect on the quality as well as the antioxidant potential of meat. Wheat germ and wheat germ oil (WGO) have been used as dietary supplements for different purposes . Wheat germ contains mainly protein (26–35%), lipids (10–15%), and minerals (4%) and major bioactive compounds like tocopherol, policosanols, and sterols . Wheat germ is the richest plant source of vitamin E. The meat quality from chickens can be improved by dietary supplementation of vitamin E, which alleviates oxidative stress .
WGO is also rich in essential fatty acids like linoleic acid and alpha linolenic acid. These are helpful in performing numerous functions like lowering the cholesterol level, and enhancing endurance. WGO also stimulates the tocopherol redox-system by changing the lipid peroxidation . In addition to vitamin E and essential fatty acids, WGO also contain Vitamin B complex and is important for chemoprevention . WGO, because of its vitamin E content, protects cells against free radicals, which negatively impact the metabolism.
ALA, widely distributed in many foods, is a short chain fatty acid and is a powerful natural antioxidant . ALA is not only an antioxidant but also a co-factor for many enzymes. ALA stimulates glucose oxidation in muscles and improves insulin sensitivity. It is also helpful to reduce the oxidative stress in tissues of different mammals . The antioxidant potential of both ALA and AT is helpful in reducing lipid oxidation in both raw and processed chicken meat and meat products [11, 12]. Antioxidants in chicken meat also reduce drip loss by lowering the post mortem pH of the meat .
The aim of the present project was to investigate the antioxidant potential of different natural antioxidants and their effect on the stability, sensory attributes, and fatty acid profile of chicken nuggets made from the meat of animals fed with different antioxidants individually or in combination.
Procurement of raw materials
All the chemicals and reagents required for the study were purchased from Sigma Aldrich (Tokyo, Japan) and Merck (Merck KGaA, Darmstadt, Germany). This research project was conducted at the National Institute of Food Science and Technology (NIFSAT) and Nutrition Research Center, University of Agriculture, Faisalabad, Pakistan. The experimental design included 6 different diets supplemented with various antioxidants, singly or in combination; control, WGO, AT, ALA, WGO + ALA and AT + ALA. Chicken nuggets were made from the leg portion of chickens fed with these diets.
Chicken nugget preparation and processing
Antioxidant enriched chicken meat was used for the preparation of nuggets by following the method described by Perlo et al. . The nuggets were stored for 45 days at −18 °C and analysis was done at 15-day intervals. The raw material for manufacturing the nuggets was weighed and cleaned according to the recipe. The recipe for the preparation of nuggets is as follows. Boneless chicken (500 g), Egg (1), oil (as required for frying), black pepper (12 g), garlic paste (1 tsp), onion (1), plain flour (120 g), bread crumbs (70 g), and salt (20 g). The control and antioxidant enriched broiler meat (leg) were washed multiple times with tap water, deboned manually and minced using an electric mincer to very fine consistency for preparing nuggets of excellent texture. The minced meat and onions were mixed in a meat mixer for 5 min, followed by the addition of all other ingredients according to the recipe and mixed using a meat mixer to obtain a uniform blend. When all the ingredients were thoroughly mixed, the mixture was spread into a thin layer (10 mm thickness) and shaped into discs of 30 mm diameter (10 ± 1 g/piece). The nuggets were dipped sequentially in plain flour and bread crumbs and fried in canola oil at 180 °C till golden brown in color.
Antioxidant status of chicken nuggets
The antioxidant status of the chicken nuggets was determined by different methods described below. The total phenolic content (TPC) in the nuggets was determined by adopting the procedure described by Senevirathne et al. . The total phenolic content of the nuggets was estimated as gallic acid equivalent (mg GAE/100 g). The nugget samples were subjected to 2,2- diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity analysis according to the procedure outlined by Brand-Williams et al. . Percentage neutralization of free radicals was assayed using DPPH and calculated using the following formula. % Neutralization = 100 × (Ablank- Asample/ Ablank). ABTS+ reducing activity of the nugget samples was measured using the method described by Erel . ABTS+ reducing activity (%) = [(Acontrol – Asample) ÷ Acontrol] × 100. The ferric reducing antioxidant power (FRAP) in the nugget samples was estimated using the method described by Arshad et al.  with some modification.
Physico-chemical analysis of nuggets
pH of the nuggets was measured with a pH meter according to the method described by Sallam et al. . Ten grams of the sample was homogenized with 50 mL distilled water and pH was measured using a digital pH meter. A hand-held tristimulus colorimeter (Color Test Meter II) was used to determine the color of the nuggets at regular storage intervals (0, 15, 30, and 45 days) by following the procedure described by Elgasim and Al-Wesali . Color was determined by placing the nuggets in a petri plate under a photocell. The water content of the nuggets was measured using electronic Hygropalm water activity meter (Model Aw-Win, Rotronic, equipped with a Karl-Fast probe) at regular storage intervals, using the method described by Cosenza et al. . The textural characteristics of nuggets were measured at different storage intervals by using a texture analyzer (Mod. TA-XT2, Stable Microsystems, surrey, UK) as described by Cardoso et al. . The nuggets were fried and compression test was performed to check the texture of the product.
Lipid stability analysis
The lipid stability of chicken nuggets was determined in terms of the amounts of thiobarbituric acid-reactive substances (TBARS) and peroxide value (POV). The amount of TBARS in the nuggets was estimated as per the guidelines of Liu et al.  and TBARS was expressed as milligrams of malondialdehyde (MDA)/kg meat. The POV of the nuggets was determined by the method outlined by Shantha and Decker  and expressed as meq peroxide/kg.
Quantification of α-lipoic acid and α-tocopherol
The ALA content in the nugget samples was measured according to the method described by Satoh et al.  with some modifications. The nugget samples were prepared according to the method described by Asghar et al.  for the determination of AT. HPLC chromatograms were obtained by using a C18 column, (250 mm × 4.6 mm, 5.0 μm), System controller SCL-10 A, water pump LC-10 AT, and flow controller valve FCV-10 AL with a mobile phase of 100% methanol at a flow rate of 1 mL/min.
Fatty acid Profile
Total fatty acids were extracted from nugget samples per the method described by Folch et al. ; this method uses an antioxidant to prevent oxidation during sample preparation and a flame ionization detector (FID). The injector temperature was 250 °C and the detector temperature was 300 °C. The column temperature program initiated the runs at 100 °C for 2 min, warmed to 170 °C at 10 °C /min, held for 2 min, warmed to 220 °C at 7.5 °C /min and then again held for 20 min to facilitate optimal separation. Results are presented as percentage of total fatty acids.
Sensory evaluation of the nuggets was carried out by a trained panel at different storage intervals (0, 15, 30, and 45 days), using a 9-point hedonic scale (9 = like extremely; 1 = dislike extremely), as per the guidelines of Meilgaard et al. . Sensory assessment for various quality attributes of nuggets, such as appearance, flavor, taste, and overall acceptability, were recorded. All evaluations were conducted by panelists trained in sensory evaluation of foods made from muscle meat. The panelists carried out the evaluation in individual booths under clear white fluorescent light in the Sensory Evaluation Laboratory of NIFSAT, University of Agriculture, Faisalabad, Pakistan. During the evaluation process, the panelists were provided unsalted crackers, mineral water, and expectorant cups to neutralize and rinse their taste receptors between different samples to facilitate rational assessment. The descriptors were rated on a scale from “0” representing the lowest score and “9” the highest. The panelists were requested to rate the product quality by scoring for the selected parameters.
The work was carried out using completely randomized design (CRD), and the data obtained for different parameters was analyzed statistically using the Statistical Package, Statistic 8.1. Levels of significance (P ≤ 0.05) were determined (ANOVA) using 2-factor factorial CRD by following the principles outlined by Steel and Torrie . The means were compared using LSD.
Results and Discussions
Antioxidant potential of chicken nuggets
The group with combination of ALA and WGO showed maximum antioxidant power (637.56 μmol/Fe+2/g meat) and TPC (140.92 mg GAE/100 g meat) compared to those in the control (571.93 μmol/Fe+2/g meat) and TPC (99.02 mg GAE/100 g meat) at 0 day of storage. The ferric reducing antioxidant power and TPC decreased as the storage intervals increased. FRAP (624.84 μmol/Fe+2/g meat) and TPC (87.62 mg GAE/100 g meat) were higher at 45th day of storage in nuggets from the group fed with WGO + ALA, whereas the values were lower in control group (559.67 μmol/Fe+2/g meat and 87.62 mg GAE/100 g meat). Different studies proved that there was positive correlation between the antioxidant activities and total phenolic contents in raw and cooked chicken meat [11, 35–37]. The results showed that the antioxidant potential was higher in the meat from the group fed with both natural antioxidant (WGO) and ALA. This finding is consistent with those of Mancini et al. , who speculated that by using antioxidants like turmeric and ascorbic acid, the antioxidant potential (DPPH, ABTS and FRAP) in rabbit burgers could be increased compared to that of the control where no antioxidants were used during storage. Banerjee et al.  showed that goat meat enriched with natural antioxidants showed higher FRAP and DPPH values than those of the control.
Physico-chemical properties of chicken nuggets
pH and color of the chicken leg meat nuggets
pH of leg nuggets with storage days
Color of leg nuggets with storage days
WGO + ALA
WGO + AT
Water activity and texture of the leg chicken nuggets during storage
WGO + ALA
WGO + AT
The results showed that the water activity in chicken nuggets from the control decreased whereas the shear force value for texture increased during storage. There was increased water activity in chicken nuggets from the group fed with antioxidants. Sohaib et al.  reported that the water activity was higher and shear force value for texture was lower in antioxidant enriched chicken nuggets. Water activity tended to decrease and texture value tended to increase with storage. These results were also supported by the Arshad et al. . Malav et al.  speculated that the decrease in shear force value observed in mutton patties enriched with antioxidant was due to the reduction in compactness because of the higher moisture content and aeration that lowers the shear force. The shear force value increased during storage because of the myofibrillar protein oxidation and the resultant increase in cross-linking and aggregation in meat [48, 49].
Thiobarbituric acid reactive substances and peroxide value of the chicken nuggets
Thiobarbituric acid reactive substances and peroxide value of the leg chicken nuggets during storage
TBARS (mg MDA/kg)
POV (meq peroxide/kg)
WGO + ALA
WGO + AT
α-Lipoic acid and α-tocopherol contents
Fatty acid profile of chicken leg nuggets
Fatty acid composition and profile in chicken nuggets
WGO + ALA
WGO + AT
fatty acids (%) in leg chicken nuggets
fatty acid profile (%) in leg chicken nuggets
Sensory evaluation of chicken nuggets during storage
It is evident from Fig. 3 that the sensory score given by the panel of judges decreased as the storage interval increased. This result is in agreement with the results of many other researchers; the appearance and color of different meat products diminished as the storage interval increased [55, 56]. Other researchers have reported that the panel of judges had a better sensory perception of the nuggets enriched in natural antioxidants . The score for flavor also decreased with storage due to the loss of volatile flavoring compounds during storage. This hypothesis was supported by Thomas et al.  and Bhat et al. . The overall acceptability also decreased during storage because of the decline in the sensory score of other parameters like appearance, flavor, and taste. This decrease in overall acceptability was confirmed by the results of Malav et al.  who reported that the overall acceptability of mutton patties decreased during storage.
It is concluded that the chicken nuggets made from the leg meat of chickens fed with diet supplemented with both WGO and ALA have better antioxidant potential as well as stability during storage. Nuggets from the group of animals fed with WGO supplemented diet showed better fatty acid profile because of the higher PUFA content in WGO. It is believed that ALA regenerates other antioxidants like AT and ascorbic acid. Therefore, nuggets from the group fed with diet containing WGO + ALA had higher content of ALA and AT because of the synergism between ALA and AT. Regarding the descriptive sensory evaluation, the overall acceptability of chicken nuggets from the animals fed with both ALA and AT was the highest according to the scores awarded by the panelists. However, this score decreased on storage.
The authors are thankful to Higher Education Commission (HEC), Govt. of Pakistan for providing financial support for this study under HEC Ph.D. indigenous fellowship program. We also extend our thanks to Directorate of Poultry Research Farm, University of Agriculture Faisalabad (UAF), Pakistan, for the use of the broiler rearing facility. The authors also extend their appreciation to the International Scientific Partnership Program (ISPP) at King Saud University, for funding this research work through ISPP# 0023.
The project was funded by Higher Education Commission, Islamabad, Pakistan.
Availability of data and materials
All data presented in the manuscript.
The contribution of each of the authors for this work was as follows. MSA carried out the research and prepared the manuscript. FMA helped in planning and executing the research. MS, FS, AI, MTN and SH helped in the manuscript preparation and provided the statistical skills. JHK reviewed the manuscript. All authors read and approved the final manuscript.
The authors have no competing interests. The authors are solely responsible for the content and writing of this manuscript.
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