Next Article in Journal
Antioxidant and Immune-Related Implications of Minerals in COVID-19: A Possibility for Disease Prevention and Management
Next Article in Special Issue
Combining Conventional Organic Solvent Extraction, Ultrasound-Assisted Extraction, and Chromatographic Techniques to Obtain Pure Betanin from Beetroot for Clinical Purposes
Previous Article in Journal
Sphingolipid Levels and Signaling via Resveratrol and Antioxidant Actions in Cardiometabolic Risk and Disease
Previous Article in Special Issue
Natural Carotenoids: Recent Advances on Separation from Microbial Biomass and Methods of Analysis
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Antioxidants
Volume 12
Issue 5
10.3390/antiox12051103
antioxidants-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Assessment of Antioxidant Stability of Meat Pâté with Allium cepa Husk Extract

by
Irina Chernukha
1,
Nadezhda Kupaeva
1,*,
Daniil Khvostov
2,
Yuliya Bogdanova
3,
Jutta Smirnova
1 and
Elena Kotenkova
1,*
1
V. M. Gorbatov Federal Research Center for Food Systems, Experimental Clinic and Research Laboratory for Bioactive Substances of Animal Origin, Talalikhina St., 26, 109316 Moscow, Russia
2
V. M. Gorbatov Federal Research Centre for Food Systems of RAS, Laboratory of Molecular Biology and Bioinformatics, Talalikhina St., 26, 109316 Moscow, Russia
3
V. M. Gorbatov Federal Research Centre for Food Systems of RAS, Department of Scientific, Applied and Technological Developments, Talalikhina St., 26, 109316 Moscow, Russia
*
Authors to whom correspondence should be addressed.
Antioxidants 2023, 12(5), 1103; https://doi.org/10.3390/antiox12051103
Submission received: 21 March 2023 / Revised: 10 May 2023 / Accepted: 14 May 2023 / Published: 16 May 2023
(This article belongs to the Special Issue Antioxidants in Food, Food Waste, and By-Product)
Versions Notes

Abstract

:
Antioxidants play a very important role in the food industry. Recently, both science and industry have shown substantial preference for natural antioxidants, including searching for antioxidant substances from natural sources without undesirable side effects. The purpose of this study was to evaluate the effect of adding Allium cepa husk extract at a volume of 68 or 34 µL/g of unsalted blanched materials to replace 34% and 17% of the beef broth, respectively, which corresponded to a total antioxidant capacity (TAC) of 44.4 or 22.2 µmol-equiv. Q/100 g meat pâté (i.e., 13.42 or 6.71 mg of quercetin/100 g meat pâté), on the quality and safety indicators of the developed meat pâté. The TAC according to a ferric reducing antioxidant power assay, thiobarbituric acid reactive substances, and physicochemical and microbiological characteristics were determined during the storage of the meat pâté. Proximal and UPLC-ESI-Q-TOF-MS analyses were also performed. The addition of yellow onion husk ethanolic extract to the meat pâté at both volumes allowed the maintenance of an increased content of antioxidants, which contributed to a decrease in the generation of secondary products of lipid peroxidation for 14 days of storage at 4 °C. The results of the microbiological analyses showed that the developed meat pâtés were safe according to all indicators of microbial spoilage within 10 days of production. The results supported the use of yellow onion husk extract in the food industry to contribute to improving the functionality of meat products, developing products for a healthy lifestyle, and providing clean-label foods without or with a minimal content of synthetic additives.

1. Introduction

Food products are important for maintaining many functions in humans, such as energy production; the supply of nutrients, including proteins and macro- and micronutrients; the maintenance of various metabolic processes; and the growth of the body [1]. A relationship exists between the food one consumes and one’s health [2], which promotes the development of functional food products that are characterized by scientifically proven biological activities and exert a beneficial physiological effect [3]. Food products can be made functional through fortification with natural (non-modified) ingredients and the addition, removal, or modification of the recipe by technological or biotechnological methods [4].
Meat and meat-containing products are one of the main food groups with high biological value, providing proteins and minerals—in particular, zinc and iron [5]. In addition, meat and meat-based products have a variety of preventive and health-promoting benefits [6], and meat is one of the most important food products along with vegetables, fruits, dairy products, and fish [7]. Therefore, meat is an acceptable matrix for functional food development [6]. However, the meat matrix is one of the most difficult for creating functional food products due to its complex composition and propensity toward oxidation and microbial spoilage [7]. The meat industry offers three options for the development of functional products: modifications at the farm level, raw materials, and meat products [8]. The beneficial properties of meat products can be improved by adding biologically active substances or probiotics, eliminating or replacing synthetic additives, and reducing cholesterol and energy levels [6,8]. Thus, a promising direction for the creation of functional meat products is the replacement of synthetic antioxidants with natural alternatives by enriching the product with plant antioxidants.
Liver and meat pâtés are cooked food products that have historically been considered traditional meals and are wide-spread in many countries, especially in Europe. The main ingredients of a pâté are liver, meat, fat, water, salt, various spices, and synthetic antioxidants [9,10]. Usually, pâtés are characterized by a high content of saturated fatty acids, fat, and non-heme iron and a low concentration of natural antioxidants. In addition, the intensive mincing of raw materials during pâté processing increases the sensitivity of lipids and proteins to oxidation [9,11]. Lipid oxidation is the main non-microbial cause of the deterioration of meat and meat products. Lipid oxidation is a very complex process involving many mechanisms interacting with each other. In brief, unsaturated fatty acids react with molecular oxygen by a free-radical mechanism [12]. During oxidative processes, hydroperoxides are generated, which are subject to further oxidation or decomposition with the formation of secondary reactive products such as aldehydes, ketones, acids, and alcohols. The presence of these compounds leads to a decrease in the quality and shelf life of meat products in terms of color, taste, aroma, texture, and nutritional value [10,13,14,15,16]. In addition, one of the most important problems of lipid oxidation is the generation of harmful compounds that could cause diseases, including atherosclerosis, cancer, inflammation, and aging processes stimulation [17,18,19]. The use of antioxidants in meat and meat-based products is the main way to slow down their oxidation in order to extend their shelf lives [10,13]. The antioxidants used in the food industry can be divided into natural and synthetic antioxidants. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and tertiary butylhydroquinone (THBQ) are the most widely used and studied [16,20], as well as ascorbic acid and tocopherol. Although both types of antioxidants play a very important role in the food industry, both science and industry have recently shown substantial preference for natural antioxidants, including searching for antioxidant substances from natural sources without undesirable side effects [21,22,23]. This trend is explained by the fact that synthetic antioxidants at high concentrations can have a negative impact on health [21,24], as well as the increase in consumer demand for “natural” products and products “without preservatives” [21].
Thus, plants, vegetables, fruits, and their co-products are a rich source of antioxidant compounds with good protective and therapeutic properties regarding socially significant diseases [13]. The use of such antioxidant compounds in the form of extracts is recommended, since purified phenolic compounds are more expensive. Moreover, extracts may have better antioxidant activity than pure compounds due to the potential synergistic effect [25]. Allium cepa L. (onion) was one of the first crops to be cultivated in the world, and it is currently one of the most popular [26], which is explained by its universal culinary use as a raw food or in various cooked forms: baked, boiled, stewed, grilled, fried, etc. [27]. In addition, onions are used in various forms of traditional medicine [28,29]. Onions have antimicrobial, antibacterial, anti-inflammatory, and antioxidant properties and present a bright taste and aroma; therefore, this plant is widely used in food and for the treatment of many diseases [30]. Numerous studies have confirmed that regular onion consumption reduces the risk of cancer, cardiovascular and neurodegenerative diseases, and osteoporosis [27,31]. The biological and medical benefits of onions are mainly associated with the high content of thiosulfinates and other organosulfur compounds. Besides well-known onions, several other species are also actively grown for culinary use, such as Allium porrum L., Allium fistulosum L., Allium ascalonicum Hort., Allium schoenoprasum L., and Allium tuberosum L. [28,29].
We previously showed that the husks of red and yellow onions are a promising and rich source of natural antioxidants, especially quercetin and its glycosides [32]. By the method of kinetic chemiluminescence, we also found that the antioxidant compounds of yellow onion husk belong to three categories, according their power. Unlike red onion husk, which had the highest total antioxidant capacity, yellow onion husk contained almost equal amounts of strong, medium, and weak antioxidants, which suggested that this extract exerted a more uniform and long-lasting antioxidant effect. In addition, we determined that the long-term consumption of yellow onion husk extract improved the antioxidant status of aging rodents [33]. The availability and cheapness of this waste support the prospect of using onion husks as a source of natural antioxidants. Despite the substantial number of available scientific papers devoted to studying the qualitative composition of onion husks and bulbs and the effectiveness of various extraction methods [34], works addressing the preservation of the husk compounds’ antioxidant properties during meat product processing are difficult to find.
Thus, the purpose of this study was to assess the stability of the antioxidant properties of yellow onion husk ethanolic extract in a meat matrix, as well as to evaluate the effect of the added extract on the quality and safety indicators of the developed meat pâté containing beef and pork.

2. Materials and Methods

2.1. Chemicals

Quercetin (purity ≥ 95%, Bangalore, Karnataka, India) and iron (III) chloride hexahydrate (purity ≥ 99%, Taufkirchen, Germany) were purchased from Sigma-Aldrich (Saint Louis, MO, USA). Dipotassium hydrogen phosphate anhydrous (purity ≥ 98%), potassium dihydrogen phosphate (purity ≥ 98%), sodium acetate anhydrous (purity ≥ 99%), hydrochloric acid (HCl, 37%), thiobarbituric acid (purity ≥ 98%), 1-butanol (purity ≥ 99.5%), and formic acid (FA, purity ≥ 98%) were purchased from PanReac AppliChem (Darmstadt, Germany). Furthermore, 2,4,4-tris(2-pyridyl)-1,3,5-triazine (TPTZ, purity ≥ 96%) was purchased from BLDpharm (Shanghai, China). Acetic acid (purity ≥ 99.8%) and ortho-phosphoric acid (purity ≥ 85%) were purchased from Component-Reaktiv (Moscow, Russia). Sodium thiosulfate standard titer was purchased from LLC Himtitry (Pereslavl-Zalessky, Russia). Starch indicator and sodium sulphate anhydrous were purchased from Himmed (Moscow, Russia). Acetic acid was purchased from Labtech (Moscow, Russia), and potassium iodide was purchased from Reahim (Moscow, Russia). Deionized water for chromatography (18 W) was obtained using a Milli-Q Merck water purification system (Millipore, Darmstadt, Germany).

2.2. Preparation of Onion Husk Extract

For the preparation of yellow onion (Allium cepa) husk ethanolic extract (OHE), onions were obtained from the supermarket, producer OOO “Agroleto”, Krasnodar, Russia. The husks were ground (particle size 5 mm or less) and soaked in 70% ethanol (60 g/900 mL) for 24 h with gentle shaking at room temperature (22 ± 2 °C). The mixture was filtered through a paper filter and kept in an airtight bottle in a refrigerator at 4 °C until use. The total antioxidant capacity (TAC) of the OHE was 6.53 ± 0.18 mmol-equiv. Q/L.

2.3. Manufacture of Meat Pâtés

Meat pâtés were produced in the Department of Scientific, Applied and Technological Developments of V. M. Gorbatov Federal Research Centre for Food Systems of RAS according to the recipes indicated in Table 1. OHE was added instead of broth at volumes of 68 µL/g for experiment 1 (E1) and 34 µL/g for experiment 1 (E2), which corresponded to a TAC values of 44.4 and 22.2 µmol-equiv. Q/100 g meat pâté, respectively (or 13.42 and 6.71 mg of quercetin/100 g meat pâté).
Raw materials were cut into 200–300 g pieces and blanched separately in water at a temperature of 95 ± 5 °C: beef liver and lean pork for 15–20 min, pork heart for 120 min, and beef flank for 40 min. Onions were peeled and ground in a Bosch MCM3501M food processor (Bosch, Škofja Loka, Slovenia) with a power of 800 W and blanched with the addition of oil in a frying pan until fully cooked. Then, the heat-treated raw meat materials and onions were individually ground in a meat grinder Hurakan HKN-12SC (Hurakan, Guangzhou, China) to a particle size of 2–3 mm and homogenized in a cutter (Robot-Coupe, Montceau-les-Mines, France) at 3000 rpm for 5 min. The ingredients were added during final homogenization in the following order: minced beef flank, lean pork, and pork heart; minced beef liver; fried onion and dry ingredients; and beef broth. This product constituted the control sample for the experimental variants. The technological variance factor, which changed the composition and quality of the final products, was the replacement of 34% (E1) and 17% (E2) of the beef broth with OHE. The final temperature of the product at the end of homogenization was over 40 °C. The product was packed at 100 ± 1 g in vacuum packaging VakumPak-M (Webomatic, Bochum, Germany), PA/PE, size 150 × 200 mm, thickness 70 µm; cooked at a temperature of 72 °C in the geometric center of the bar; and then cooled to 4 °C in a water bath (EKROS 4310, Saint Petersburg, Russia) for 20–30 min and stored at 4 °C. The temperature control was carried out using a digital thermometer WT-1 (Xuzhou Sanhe Automatic Control Equipment Co., Ltd., Xuzhou, China). Samples were periodically taken for analyses after 1, 3, 5, 7, 10, 14, or 28 days of storage, depending on the studied indicators.
In order to investigate the influence of OHE on the quality, safety, and chemical composition of the pâté, the following measurements were carried out: TAC and thiobarbituric acid reactive substances (TBARS) (0, 3, 5, 7, and 14 days of storage), and physicochemical and microbiological characteristics (0, 3, 7, 10, 14, and 28 days of storage). Proximal and UPLC-ESI-Q-TOF-MS analyses were performed on day 0 of storage.

2.4. Proximal Analysis

The techniques employed included the Kjeldahl method for protein, the Soxhlet method with acid hydrolysis for fat, and drying- and vacuum-oven methods for moisture and ash assessment based on the Association of Official Analytical Chemists (AOAC): Official Methods of Analysis [35]; total carbohydrates were calculated by their difference.

2.5. Extraction of Meat Pâtés

To determine the TAC and perform UPLC-ESI-Q-TOF-MS analysis, ethanolic extracts of the meat pâtés were prepared, and phosphate extracts were prepared to measure the TBARS. The sample was mixed with 96% ethanol or 50 mM phosphate buffer (pH 7.0) in a ratio of 1:5 (g:mL) and homogenized using an S10 manual homogenizer (Stegler, Guangzhou, China) for 2 min at 9000 rpm. Phosphate extracts were centrifuged at 7000× g for 5 min at 4 °C in a 5427R centrifuge (Eppendorf AG, Hamburg, Germany); ethanolic extracts were infused for 60 min at 22 ± 2 °C, followed by filtration through a paper filter. The obtained extracts were stored at minus 40 °C.

2.6. UPLC-ESI-Q-TOF-MS Analysis

The metabolome analysis of meat pâtés and OHE was performed using an UHPLC 1290 Infinity system (Agilent Technologies, Santa Clara, CA, USA), as described previously [36], with some modifications. Analysis was performed using a Luna Omega C18 analytical column (2.1 mm × 50 mm, 1.6 µm particle size, Phenomenex Inc., Torrance, CA, USA). The column temperature was maintained at 60 °C, the injection volume was 5 µL, and the linear gradient was as follows: 0% solvent B for 2 min, progression from 0% to 85% solvent B for 8 min, and 85% solvent B for 2 min. The total analysis time was 15 min. An Agilent 6545XT AdvanceBio LC/QTOF (Agilent Technologies, Santa Clara, CA, USA), set to positive ionization mode, was used for the high-pressure ion funnel. The capillary voltage was 4500 V; the nozzle voltage was 2000 V; the drying gas flow was operated at 8 L/min and 325 °C; the gas flow through the casing was operated at 12 L/min and 275 °C; and the atomizer pressure was 30 psi, with a high frequency (RF) of 175 V.
Detected compounds were identified by MS fragmentation using MSDIAL software (ver. 5.1.221218, RIKEN CSRS, Yokohama City, Japan) [37]. The total score for manually selected compounds was ≥80%. Flavonoid contents were determined according to a standard curve using quercetin (Q) in the concentration range of 1–1000 ng/mL [32] and expressed in µg-eq. Q/100 mL OHE or µg-eq. Q/100 g meat pâté.

2.7. Stability Assessment

TAC and TBARS were measured after 0, 3, 5, 7, and 14 days of storage at 4 °C; peroxide value (PV), pH, and microbiological parameters were determined after 3, 7, 10, 14, and 28 days of storage at 4 °C.

2.7.1. Ferric Reducing Antioxidant Power (FRAP) Assay

The TAC was measured by the FRAP method using an SF-2000 spectrophotometer (OCB Spectr, St. Petersburg, Russia) according to [38], with some modifications [33]. In brief, 1.45 mL of FRAP reagent and 50 μL of the sample/standard/distilled water (control) were mixed and incubated for 30 min at 37 °C in the dark. The optical density was determined at 594 nm. The standard curve of quercetin (Q) in the concentration range of 140–300 μM was used. The results are expressed in mmol-equiv. Q/L OHE or µmol-equiv. Q/100 g meat pâté.

2.7.2. Lipid Peroxidation Products

The TBARS in the phosphate extracts of the meat pâtés were measured using an SF-2000 spectrophotometer following the method of Brazhe et al. [39], with some modifications [33]. In brief, glass tubes were filled with 1.5 mL of 2% (w/v) ortho-phosphoric acid, 100 µL of extract or distilled water for the control sample, and 0.5 mL of 0.8% (w/v) thiobarbituric acid. After incubation at 95 °C for 45 min, samples were cooled to room temperature; then, 2.5 mL of n-butanol was added, and the samples were mixed and centrifuged. Optical density was measured at wavelengths of 535 and 570 nm. TBARS were calculated using the molar extinction coefficient of the (malondialdehyde) MDA-TBA complex (1.56 × 105 M−1 cm−1) and expressed in µmol/100 g meat pâté.

2.7.3. Determination of Peroxide Value and pH

PV was determined according to the ISO 3960:2017 standard [40] based on the reaction of fat oxidation products (peroxides and hydroperoxides) with potassium iodide in an acidic medium. A solution of sodium thiosulfate was used for titration following the quantitative determination of the released iodine. The results are expressed as mmol. active O2/kg of fat. The pH was measured according to the ISO 2917:1999 standard [41] using a FiveEasy FP20 (Mettler Toledo, Stockholm, Sweden).

2.7.4. Microbiological Analyses

The following microbial parameters were determined: total mesophilic aerobic bacteria (TMAB), according to the ISO 4833-1:2013 standard [42]; sulfite-reducing bacteria growing under anaerobic conditions, according to the ISO 15213:2003 standard [43]; yeasts and molds, according to the ISO 21527-2:2008 standard [44]; Escherichia coli, according to the ISO 16654:2001 standard [45]; coliform bacteria, according to the ISO 4832:2006 standard [46]; Salmonella spp., according to the ISO 6579:2002 standard [47]; Bacillus cereus, according to the ISO 7932:2004 standard [48]; presumptive Pseudomonas spp., according to the ISO 13720:2010 standard [49]; coagulase-positive staphylococci (Staphylococcus aureus and other species), according to the ISO 6888-1:2021 standard [50]; and Listeria monocytogenes and Listeria spp., according to the ISO 11290-1:2017 standard [51].

2.8. Statistical Analyses

The measurements were carried out in triplicate. STATISTICA 17.0 software was used for the statistical analysis. The results were calculated as mean ± SD. Significant differences were tested by non-parametric Mann–Whitney U tests for independent variables; Freidman ANOVAs (n > 2) were used for dependent variables. Differences with p-values < 0.10 and 0.05, respectively, were considered statistically significant. After processing the UPLC-ESI-Q-TOF-MS data using the MS-DIAL program (version 5.1.221218, RIKEN CSRS, Yokohama, Japan), the metabolomic peaks were identified, including the collection of peaks, deconvolution, the identification of compounds, and the alignment of peaks to a reference database [37].

3. Results

3.1. Composition of Meat Pâtés

The physico-chemical composition of the meat pâtés is presented in Table 2.
E1 was characterized by a higher content of protein and ash, statistically exceeding the control indicators by 1.32-fold and 1.08-fold (p < 0.10), respectively, while the content of fat and moisture was lower by 1.33% and 4.17% (p < 0.10), respectively. E2 demonstrated the same tendency according to the proximal analysis, but the difference between E2 and the control was not as substantial. A decrease in the fat and moisture content accompanied an elevation in the protein content as follows: control > E2 > E1.

3.2. Identification of Active Compounds and Metabolome Profile in Meat Pâtés

Ethanolic extracts of the meat pâtés and OHE were investigated by UPLC-ESI-Q-TOF-MS analysis. More than 100 compounds were obtained using the MSDIAL accurate mass tolerance MS1 (0.01 Da) and MS2 (0.05 Da) program parameters. Table S1 (Supplementary Materials) shows the mass parameters and identification characteristics of all manually selected compounds for all samples. A total of 69 compounds were manually selected, including phosphoethanolamines (n = 4); other lipids (n = 10); acyl carnitines (n = 5); alpha amino acids and derivatives (n = 5); B vitamins and related compounds (n = 5); benzodioxoles (n = 5); flavonoids (n = 10); trihydroxy bile acids, alcohols, and derivatives (n = 4); and other organic and polyphenolic compounds (n = 21).
Table 3 shows the main tentative compounds determined in the meat pâtés and OHE; the chromatograms and spectra of the quercetin in the studied samples are presented in Figure S1 (Supplementary Materials); and the ID/structure, representative mass spectra compared to the reference, and total score for each compound are presented in Table S2 (Supplementary Materials). The main flavonoids were quantitatively determined using the calibration curve of quercetin; the regression coefficient was >0.990.
The predicted values of the compounds in the E1 and E2 meat pâtés were calculated based on the OHE results and the volumes added to the recipes for E1 and E2 (Table 1), and they were expected to correspond to ∆ (E1-C) and ∆ (E2-C). Regular spices were added to the recipes of both the control and experimental meat pâtés, which were also a source of antioxidants; therefore, the values of the control sample were subtracted from the experimental values.
The total amount of flavonoids in E1 and E2 was significantly higher than in the control by 2.8-fold (p < 0.1) and 1.9-fold (p < 0.1), respectively, while the difference between E1 and E2 averaged 1.5-fold (p < 0.1). The content of all flavonoids in E1 and E2 exceeded the control values, except for petunidin 3-galactoside. Thus, the concentrations of delphinidin 3-galactoside, luteolin-4′-O-glucoside, spiraeoside, myricitrin, isorhamnetin-3-O-beta-D-Glucoside, isorhamnetin, kaempferol, and quercetin in E1 were greater than in the control by 4.3-fold (p < 0.1), 2.0-fold (p < 0.1), 1.9-fold (p < 0.1), 2.6-fold (p < 0.1), 1.4-fold (p < 0.1), 6.0-fold (p < 0.1), 18.0-fold (p < 0.1), and 131.9-fold (p < 0.1), respectively, while for E1 these differences were 1.9-fold (p < 0.1), 1.4-fold (p < 0.1), 1.5-fold (p < 0.1), 2.0-fold (p < 0.1), 1.5-fold (p < 0.1), 3.3-fold (p < 0.1), 7.4-fold (p < 0.1), and 58.5-fold (p < 0.1), respectively. Quercetin 3-O-malonylglucoside, quercetin-3,4′-O-di-beta-glucoside, and baimaside presented in trace amounts in the control, while the content of these compounds in E1 and E2 was in the range of 8.37–73.66 µg-eq. Q/100 g meat pâté. The content of all flavonoids in E1, except for petunidin 3-galactoside and isorhamnetin-3-O-beta-D-Glucoside, exceeded the E2 values by 1.3–2.4-fold (p < 0.1).
Based on the determination of the main compounds in OHE, the predicted content in E1, E2, ∆ (E1-C), and ∆ (E2-C) was calculated in order to evaluate the antioxidant stability of Allium cepa husk components during meat pâté processing. We found that the content in ∆ (E1-C) and ∆ (E2-C) was lower than predicted. However, the difference between ∆ (E1-C) and ∆ (E2-C) averaged 1.9-fold, which corresponded to the ratio of OHE volume in E1 and E2.

3.3. Determination of Antioxidant Stability

The results of the TAC determination for the meat pâtés using the FRAP method after 0, 3, 5, 7, and 14 days of storage at 4 °C are presented in Table 4.
During storage, a decrease in the TACFRAP was observed in all samples. The TACFRAP of E1 and E2 for all storage durations statistically exceeded the indicators for the control meat pâté. Thus, the TACFRAP of E1, with an ethanolic OHE volume of 68 µL/g of raw materials, exceeded that of the control pâté by 36.67 ± 5.86 µmol-equiv. Q/100 g meat pâté, whereas the TACFRAP of E2, with an OHE volume of 34 µL/g exceeded the control value by 16.39 ± 2.75 µmol-equiv. Q/100 g meat pâté. We determined that a 50% decrease in the volume of OHE in the meat product recipe led to a statistically significant reduction in the contribution of the plant extract to the TACFRAP. On day 0, the difference between the TACFRAP of E1 and E2, as well as the ∆ values, was about 2.3 times.
The decreases in the TACFRAP of the OHE during meat product processing were calculated and are presented in Table 5.
When more OHE was added to the recipe, greater decreases in the TACFRAP were observed. Thus, the decrease for E1 averaged 17.41%, while it averaged only 9.45% for E2.

3.4. Determination of Storage Indicators

The results of the PV, pH, TBARS, and microbiological parameter determination are presented in Table 6, Table 7 and Table 8.
The PV of all samples increased by statistically significant amounts over the 28 days of storage (p < 0.05) but did not change significantly during the first 7 days (p > 0.05). The PV of E2 was the smallest on day 0, being significantly lower than that of the control and E1 by 0.72 and 0.74 mmol. active O2/kg of fat (p < 0.10), respectively. However, on day 3 of storage, E2 achieved the highest PV, which was 0.57 mmol. active O2/kg of fat (p < 0.10) higher than that of the control but did not differ from that of E1. After 7 days of storage, the PV of E2 was statistically lower than that of the control and E1, and after 10 days it was higher than that of E1 but lower than that of the control.
The pH value changed by statistically significant amounts in the control and E1 over the 28 days of storage (p < 0.05) but did not change significantly in any sample during the first 7 days of storage. On day 0 and 3 of storage, the pH of E1 did not differ from that of the control, while the pH of E2 did not differ from that of the control on day 0 alone. On day 0, the pH of E2 was lower than that of E1 and the control, but after 3 days of storage E2 demonstrated the highest pH value, which statistically differed from that of the control and E1. Despite the observed differences, the pH of the meat pâtés varied in the range of 5.83–6.07, representing an insignificant change for this type of product.
The TBARS of the meat pâtés did not differ significantly on day 0. However, the TBARS in the control were slightly increased by about 0.25 µmol/100 g meat pâté. After 3, 5, 7, and 14 days of storage, we found a significant difference in the TBARS between the control and experimental samples, while we observed no statistical difference between the TBARS of E1 and E2. On day 3 of storage, the concentration of TBARS in the control exceeded that in E1 and E2 by 2.42-fold and 2.1-fold (p < 0.10), respectively. During storage, a statistical change in TBARS was observed for the control and E2 (p < 0.05), whereas the TBARS in E1 changed only slightly. The TBARS in E1 remained unchanged for 5 days, while those in E2 remained unchanged for 3 days.
Table S3 (Supplementary Materials) shows the results of the microbiological analyses of the meat pâtés during storage at 4 °C, showing that the developed meat pâtés were safe according to all indicators of microbial spoilage within 10 days of production.

4. Discussion

Recently, a trend of replacing synthetic antioxidants with natural ones has emerged, including for the purpose of enriching foods with essential nutrients [52]. Antioxidants from natural sources are a good alternative to synthetic antioxidants due to their high content of phenols and other active components that can effectively prevent lipid oxidation [24]. Plant extracts are becoming important additives in the food industry due to their antimicrobial and antioxidant properties, which delay the development of undesirable tastes and improve the color stability of meat products [21]. Therefore, the use of antioxidants in the meat industry is a reasonable and necessary step to extend the shelf lives and maintain the organoleptic and nutritional qualities of meat products. From this point of view, the use of plant extracts in the food industry contributes to improving the functionality of meat products, developing products for a healthy lifestyle, and manufacturing clean-label foods without or with a minimal content of synthetic additives [21,53,54,55].
Numerous studies have shown the effectiveness of using plant extracts or ingredients to slow down oxidative processes in meat products. Monica Gallo et al. demonstrated the effectiveness of Echinacea angustifolia extract for reducing the oxidation of lipids and proteins in chicken meat [16]. Mario Estévez et al. slowed down the lipid peroxidation in liver pâté through the addition of sage and rosemary essential oils [11]. Lilian Regina Barros Mariutti et al. demonstrated that the addition of sage to chicken meat (0.1 g/100 g) is a good alternative to prevent and slow down the formation of compounds as a result of lipid oxidation that are responsible for unpleasant tastes and a loss of nutritional qualities during prolonged freezer storage [56]. Pellegrini et al. assessed the effect of partial fat replacement with quinoa paste on the quality of pork liver pâté [9]. The authors found that replacing 10% of the fat lowered the oxidation rate of the product in comparison to that of the control sample. An equally promising source of plant antioxidants is the waste from the fruit and vegetable industry. This area is currently under study, and biologically active substances obtained from such waste have been used as antioxidants. Jose M. Lorenzo et al. demonstrated the possibilities of using peanut skins, which are a by-product of peanut production, as a source of antioxidant compounds, especially proanthocyanidins, which are capable of inhibiting oxidative reactions involving mainly pigments, lipids, and proteins [57]. Our results demonstrated that the addition of yellow onion husk ethanolic extract to meat pâté at a volume of 68 µL/g or 34 µL/g unsalted blanched materials to replace 34% or 17% of the beef broth, respectively, corresponding to a TACFRAP value of 44.4 or 22.2 µmol-equiv. Q/100 g meat pâté, respectively, (i.e., 13.42 or 6.71 mg of quercetin/100 g meat pâté) allowed us to maintain an increased content of antioxidants, which decreased the generation of secondary products of lipid peroxidation for 14 days at 4 °C. However, the TACFRAP of both the control and experimental samples declined during storage, which was explained by the inhibition of oxidation products by antioxidants from the spices and OHE. This observation was confirmed by the increases in the PV and TBARS during storage. Remarkably, the increases in the lipid oxidation indicators in the experimental samples were not as noticeable as those in the control samples. We also showed that the addition of 34 µL/g of the extract to the recipe better preserved the effectiveness of the antioxidants. Thus, for their entire shelf life, a TAC decrease of 33.93% was noted for the control sample, 31.27% for E1, and only 27.60% for E2.
The control meat pâté also demonstrated a prominent TACFRAP and, according to the results of the UPLC-ESI-Q-TOF-MS analysis, contained almost all the detected flavonoids. Regular spices were included in the recipes for both the control and experimental meat pâtés, such as black pepper, allspice, mustard, and nutmeg, which are also sources of antioxidants. Black pepper contains flavonoids such as catechin, quercetin, myricetin, carotenoids [58], kaempferol 3-O-glucosyl-rhamnosyl-galactoside myricetin 3-O-rhamnoside, kaempferol 3-O-(2″-rhamnosyl-galactoside) 7-O-rhamnoside, isorhamnetin 3-O-glucoside7-O-rhamnoside, and delphinidin 3-O-glucosyl-glucoside [59], and its content of total flavonoids could reach 2149 μg QE/100 g [60]. Allspice also contains flavonoids, such as quercetin [61], gallic acid, quercetin 3-O-β-D-galactoside, quercetin 3-O-β-D-glucuronide 6″-methyl ester, myricetin, myricetin 3-O-β-D-galactoside, kaempferol 3-O-β-D-glucoside, and kaempferol 3-O-β-D-galactoside, which were isolated by repeated column chromatography from dried ground berries of allspice in [62]. A total of 26 compounds (phenolic acids, flavonoids, and glucosinolates) were identified in mustard samples [63], and among the flavonoids, the most abundant were quercetin and epicatechin [64]. Flavones, flavonols, and anthocyanidins appear to be more abundant in nutmeg [65]. Nutmeg could contain saponin, alkaloid, tannin, flavonoids, α-pinene, β-pinene, myrcene, 1,8-cineole, carvacrol, terpinen-4-ol, sabinene, camphene, myristicin, elemicin, isoelemicin, eugenol, etc. [66,67]. Although the presence of almost all of the detected flavonoids in the control meat pâté could be explained by the regular spices included in the recipe, surprisingly, spiraeoside was the most abundant antioxidant in this sample, averaging 11,041.1 ± 158.1 µg-eq. Q/100 g meat pâté. Spiraeoside (quercetin 3,4′-diglucoside) is one of the predominant flavonoids present in the husk of Allium cepa L. [68], alongside quercetin (the most abundant in onion husks) and its glycosides, kaempferol and myricetin [32,69,70,71,72]. However, these are not abundant in black pepper, allspice, mustard, or nutmeg. Fried onions were included in the meat pâté recipes. Despite our previous results showing that the total antioxidant capacity of onion bulbs is much lower than that of onion husks [73], as well as data reporting that the quercetin content in onion husks is about 32 times higher than in the edible parts of onions [70], quercetin 3,4′-O-diglucoside was detected as a major flavonol in onion bulbs [74], and yellow onion bulbs tend to accumulate quercertin-3,4′-diglucoside [75]. However, in this study we demonstrated that the flavonoid content in the experimental meat pâtés was significantly higher than in the control samples. Moreover, the addition of onion husk extract in both volumes during meat pâté processing ensured the presence of significant amounts of quercetin, the main flavonoid in onion husks, in developed products in appropriate ratio.

5. Conclusions

We found that the introduction of ethanolic yellow onion husk extract into the meat matrix at a volume of 68 µL/g of raw materials reduced the indicators of oxidative spoilage and increased the total antioxidant capacity by more than 1.5 times, which was maintained for 14 days of storage at 4 °C. A reduction in the amount of added ethanolic yellow onion husk extract ensured the preservation of the antioxidant effect in the meat matrix for 14 days of storage, allowing a decrease in the concentration of ethanol in the developed meat pâté. Our results support the use of yellow onion husk extract in the food industry to contribute to improving the functionality of meat products, developing products for a healthy lifestyle, and providing clean-label foods without or with a minimal content of synthetic additives.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/antiox12051103/s1, Table S1: Mass parameters and identification characteristics of all manually selected compounds, Figure S1: Chromatogram and spectrum of quercetin in the studied samples, Table S2: Results of microbiological analyses of meat pâtés during storage at 4 °C. Table S3. Results of microbiological analyses of meat pâtés during storage at 4 °C.

Author Contributions

Conceptualization, I.C., N.K. and E.K.; methodology, N.K., J.S., Y.B. and D.K.; software, N.K. and D.K.; validation, I.C., N.K. and E.K.; formal analysis, N.K. and E.K.; investigation, N.K., Y.B., J.S. and D.K.; resources, I.C. and E.K.; data curation, I.C. and E.K.; writing—original draft preparation, I.C., N.K., D.K. and E.K.; writing—review and editing, I.C. and E.K.; visualization, N.K. and D.K.; supervision, E.K.; project administration, I.C. and E.K. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by state assignment of the V.M. Gorbatov Federal Research Center for Food Systems, No. FNEN-2019-0008.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data are contained within the article or Supplementary Materials.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Kaur, S.; Das, M. Functional foods: An overview. Food Sci. Biotechnol. 2011, 20, 861. [Google Scholar] [CrossRef]
  2. Milner, J.A. Functional foods: The US perspective. Am. J. Clin. Nutr. 2000, 71, 1654S–1659S; discussion 1674S–1675S. [Google Scholar] [CrossRef] [PubMed]
  3. Kausar, T.; Hanan, E.; Ayob, O.; Praween, B.; Azad, Z. A review on functional ingredients in red meat products. Bioinformation 2019, 15, 358–363. [Google Scholar] [CrossRef] [PubMed]
  4. Olmedilla-Alonso, B.; Jiménez-Colmenero, F.; Sánchez-Muniz, F.J. Development and assessment of healthy properties of meat and meat products designed as functional foods. Meat Sci. 2013, 95, 919–930. [Google Scholar] [CrossRef]
  5. Manassi, C.F.; de Souza, S.S.; Hassemer, G.S.; Sartor, S.; Lima, C.M.G.; Miotto, M.; De Dea Lindner, J.; Rezzadori, K.; Pimentel, T.C.; Ramos, G.L.P.A.; et al. Functional meat products: Trends in pro-, pre-, syn-, para- and post-biotic use. Food Res. Int. 2022, 154, 111035. [Google Scholar] [CrossRef] [PubMed]
  6. Ravani, A.; Sharma, H.P. Chapter 7. Meat Based Functional Foods. In Functional Foods, 1st ed.; Chhikara, N., Panghal, A., Chaudhary, G., Eds.; Scrivener Publishing LLC: Beverly, MA, USA, 2022; pp. 235–287. [Google Scholar] [CrossRef]
  7. Nazarova, N.E.; Lazutina, A.L.; Lebedeva, T.E.; Batsyna, Y.V.; Statuev, A.A. The use of plant raw materials in the production of meat pate. IOP Conf. Ser. Earth Environ. Sci. 2022, 1052, 012063. [Google Scholar] [CrossRef]
  8. Augustyńska-Prejsnar, A.; Ormian, M.; Sokołowicz, Z.; Kačániová, M. The Effect of the Addition of Hemp Seeds, Amaranth, and Golden Flaxseed on the Nutritional Value, Physical, Sensory Characteristics, and Safety of Poultry Pâté. Appl. Sci. 2022, 12, 5289. [Google Scholar] [CrossRef]
  9. Pellegrini, M.; Lucas-Gonzalez, R.; Sayas-Barberá, E.; Fernández-López, J.; Pérez-Álvarez, J.A.; Viuda-Martos, M. Quinoa (Chenopodium quinoa Willd) paste as partial fat replacer in the development of reduced fat cooked meat product type pâté: Effect on quality and safety. CyTA J. Food 2018, 16, 1079–1088. [Google Scholar] [CrossRef]
  10. Doolaege, E.H.; Vossen, E.; Raes, K.; De Meulenaer, B.; Verhé, R.; Paelinck, H.; De Smet, S. Effect of rosemary extract dose on lipid oxidation, colour stability and antioxidant concentrations, in reduced nitrite liver pâtés. Meat Sci. 2012, 90, 925–931. [Google Scholar] [CrossRef]
  11. Estévez, M.; Ramírez, R.; Ventanas, S.; Cava, R. Sage and rosemary essential oils versus BHT for the inhibition of lipid oxidative reactions in liver pâté. LWT Food Sci. Technol. 2007, 40, 58–65. [Google Scholar] [CrossRef]
  12. Domínguez, R.; Pateiro, M.; Gagaoua, M.; Barba, F.J.; Zhang, W.; Lorenzo, J.M. A Comprehensive Review on Lipid Oxidation in Meat and Meat Products. Antioxidants 2019, 8, 429. [Google Scholar] [CrossRef] [PubMed]
  13. Echegaray, N.; Gómez, B.; Barba, F.J.; Franco, D.; Estévez, M.; Carballo, J.; Marszałek, K.; Lorenzo, J.M. Chestnuts and by-products as source of natural antioxidants in meat and meat products: A review. Trends Food Sci. Technol. 2018, 82, 110–121. [Google Scholar] [CrossRef]
  14. Fernandes, R.P.P.; Trindade, M.A.; Tonin, F.G.; Pugine, S.M.P.; Lima, C.G.; Lorenzo, J.M.; de Melo, M.P. Evaluation of oxidative stability of lamb burger with Origanum vulgare extract. Food Chem. 2017, 233, 101–109. [Google Scholar] [CrossRef]
  15. Martín-Sánchez, A.M.; Ciro-Gómez, G.; Sayas, E.; Vilella-Esplá, J.; Ben-Abda, J.; Pérez-Álvarez, J.Á. Date palm by-products as a new ingredient for the meat industry: Application to pork liver pâté. Meat Sci. 2013, 93, 880–887. [Google Scholar] [CrossRef]
  16. Gallo, M.; Ferracane, R.; Naviglio, D. Antioxidant addition to prevent lipid and protein oxidation in chicken meat mixed with supercritical extracts of Echinacea angustifolia. J. Supercrit. Fluids 2012, 72, 198–204. [Google Scholar] [CrossRef]
  17. Pereira, A.L.F.; Abreu, V.K.G. Lipid peroxidation in meat and meat products. In Lipid Peroxidation, 1st ed.; Mansour, M.A., Ed.; IntechOpen: London, UK, 2018; pp. 29–42. [Google Scholar] [CrossRef]
  18. Alfaia, C.M.; Alves, S.P.; Lopes, A.F.; Fernandes, M.J.; Costa, A.S.; Fontes, C.M.; Castro, M.L.; Bessa, R.J.; Prates, J.A. Effect of cooking methods on fatty acids, conjugated isomers of linoleic acid and nutritional quality of beef intramuscular fat. Meat Sci. 2010, 84, 769–777. [Google Scholar] [CrossRef] [PubMed]
  19. Broncano, J.M.; Petrón, M.J.; Parra, V.; Timón, M.L. Effect of different cooking methods on lipid oxidation and formation of free cholesterol oxidation products (COPs) in Latissimus dorsi muscle of Iberian pigs. Meat Sci. 2009, 83, 431–437. [Google Scholar] [CrossRef]
  20. Shah, M.A.; Bosco, S.J.; Mir, S.A. Plant extracts as natural antioxidants in meat and meat products. Meat Sci. 2014, 98, 21–33. [Google Scholar] [CrossRef]
  21. Nikmaram, N.; Budaraju, S.; Barba, F.J.; Lorenzo, J.M.; Cox, R.B.; Mallikarjunan, K.; Roohinejad, S. Application of plant extracts to improve the shelf-life, nutritional and health-related properties of ready-to-eat meat products. Meat Sci. 2018, 145, 245–255. [Google Scholar] [CrossRef]
  22. Agregán, R.; Lorenzo, J.M.; Munekata, P.E.S.; Dominguez, R.; Carballo, J.; Franco, D. Assessment of the antioxidant activity of Bifurcaria bifurcata aqueous extract on canola oil. Effect of extract concentration on the oxidation stability and volatile compound generation during oil storage. Food Res. Int. 2017, 99, 1095–1102. [Google Scholar] [CrossRef]
  23. Franco, D.; Rodríguez-Amado, I.; Agregán, R.; Munekata, P.E.S.; Vázquez, J.A.; Barba, F.J.; Lorenzo, J.M. Optimization of antioxidants extraction from peanut skin to prevent oxidative processes during soybean oil storage. LWT 2018, 88, 1–8. [Google Scholar] [CrossRef]
  24. Kumar, Y.; Yadav, D.N.; Ahmad, T.; Narsaiah, K. Recent Trends in the Use of Natural Antioxidants for Meat and Meat Products. Compr. Rev. Food Sci. Food Saf. 2015, 14, 796–812. [Google Scholar] [CrossRef]
  25. Calliste, C.A.; Trouillas, P.; Allais, D.P.; Duroux, J.L. Castanea sativa Mill. leaves as new sources of natural antioxidant: An electronic spin resonance study. J. Agric. Food. Chem. 2005, 53, 282–288. [Google Scholar] [CrossRef] [PubMed]
  26. Fredotović, Ž.; Šprung, M.; Soldo, B.; Ljubenkov, I.; Budić-Leto, I.; Bilušić, T.; Čikeš-Čulić, V.; Puizina, J. Chemical Composition and Biological Activity of Allium cepa L. and Allium × cornutum (Clementi ex Visiani 1842) Methanolic Extracts. Molecules 2017, 22, 448. [Google Scholar] [CrossRef]
  27. González-de-Peredo, A.V.; Vázquez-Espinosa, M.; Espada-Bellido, E.; Ferreiro-González, M.; Carrera, C.; Barbero, G.F.; Palma, M. Development of Optimized Ultrasound-Assisted Extraction Methods for the Recovery of Total Phenolic Compounds and Anthocyanins from Onion Bulbs. Antioxidants 2021, 10, 1755. [Google Scholar] [CrossRef] [PubMed]
  28. Corzo-Martínez, M.; Corzo, N.; Villamiel, M. Biological properties of onions and garlic. Trends Food Sci. Technol. 2007, 18, 609–625. [Google Scholar] [CrossRef]
  29. Lanzotti, V. The analysis of onion and garlic. J. Chromatogr. A 2006, 1112, 3–22. [Google Scholar] [CrossRef]
  30. Arshad, M.S.; Sohaib, M.; Nadeem, M.; Saeed, F.; Imran, A.; Javed, A.; Amjad, Z.; Batool, S.M. Status and trends of nutraceuticals from onion and onion by-products: A critical review. Cogent Food Agric. 2017, 3, 1. [Google Scholar] [CrossRef]
  31. Law, Y.Y.; Chiu, H.F.; Lee, H.H.; Shen, Y.C.; Venkatakrishnan, K.; Wang, C.K. Consumption of onion juice modulates oxidative stress and attenuates the risk of bone disorders in middle-aged and post-menopausal healthy subjects. Food Funct. 2016, 7, 902–912. [Google Scholar] [CrossRef]
  32. Chernukha, I.; Kupaeva, N.; Kotenkova, E.; Khvostov, D. Differences in Antioxidant Potential of Allium cepa Husk of Red, Yellow, and White Varieties. Antioxidants 2022, 11, 1243. [Google Scholar] [CrossRef]
  33. Chernukha, I.; Fedulova, L.; Vasilevskaya, E.; Kulikovskii, A.; Kupaeva, N.; Kotenkova, E. Antioxidant effect of ethanolic onion (Allium cepa) husk extract in ageing rats. Saudi J. Biol. Sci. 2021, 28, 2877–2885. [Google Scholar] [CrossRef] [PubMed]
  34. Liu, Z.-W.; Zeng, X.-A.; Ngadi, M. Enhanced extraction of phenolic compounds from onion by pulsed electric field (PEF). J. Food Process. Preserv. 2018, 42, e13755. [Google Scholar] [CrossRef]
  35. The Association of Official Analytical Chemists (AOAC). Official Methods of Analysis, 17th ed.; The Association of Official Analytical Chemists (AOAC): Gaithersburg, MD, USA, 2000. [Google Scholar]
  36. Chernukha, I.; Kotenkova, E.; Derbeneva, S.; Khvostov, D. Bioactive Compounds of Porcine Hearts and Aortas May Improve Cardiovascular Disorders in Humans. Int. J. Environ. Res. Public Health 2021, 18, 7330. [Google Scholar] [CrossRef]
  37. Tsugawa, H.; Nakabayashi, R.; Mori, T.; Yamada, Y.; Takahashi, M.; Rai, A.; Saito, K. A cheminformatics approach to characterize metabolomes in stable-isotope-labeled organisms. Nat. Methods 2019, 16, 295–298. [Google Scholar] [CrossRef]
  38. Benzie, I.F.F.; Strain, J.J. The ferric reducing ability of plasma (FRAP) as a measure of ‘antioxidant power’: The FRAP assay. Anal. Biochem. 1996, 239, 70–76. [Google Scholar] [CrossRef] [PubMed]
  39. Brazhe, N.A.; Baizhumanov, A.A.; Parshina, E.Y.; Yusipovich, A.I.; Akhalaya, M.Y.; Yarlykova, Y.V.; Maksimov, G.V. Studies of the blood antioxidant system and oxy-gen-transporting properties of human erythrocytes during 105-day isolation. Hum. Physiol. 2014, 40, 804–809. [Google Scholar] [CrossRef]
  40. ISO 3960:2017; Animal and Vegetable Fats and Oils—Determination of Peroxide Value—Iodometric (Visual) Endpoint Determination. ISO: Geneva, Switzerland, 2017. Available online: https://www.iso.org/standard/71268.html (accessed on 1 November 2022).
  41. ISO 2917:1999; Meat and Meat Products—Measurement of pH. ISO: Geneva, Switzerland, 1999. Available online: https://www.iso.org/ru/standard/24785.html (accessed on 1 November 2022).
  42. ISO 4833-1:2013; Microbiology of the Food Chain—Horizontal Method for the Enumeration of Microorganisms—Part 1: Colony Count at 30 °C by the Pour Plate Technique. ISO: Geneva, Switzerland, 2013. Available online: https://www.iso.org/obp/ui/#iso:std:iso:4833:-1:ed-1:v1:en (accessed on 1 November 2022).
  43. ISO 15213:2003; Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Sulfite-Reducing Bacteria Growing under Anaerobic Conditions. ISO: Geneva, Switzerland, 2003. Available online: https://www.iso.org/ru/standard/26852.html (accessed on 1 November 2022).
  44. ISO 21527-2:2008; Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Yeasts and Moulds—Part 2: Colony Count Technique in Products with Water Activity Less than or Equal to 0.95. ISO: Geneva, Switzerland, 2008. Available online: https://www.iso.org/obp/ui#iso:std:iso:21527:-2:ed-1:v1:en (accessed on 1 November 2022).
  45. ISO 16654:2001; Microbiology-Horizontal Method for the Detection of Escherichia coli O157. ISO: Geneva, Switzerland, 2001. Available online: https://www.iso.org/standard/29821.html (accessed on 1 November 2022).
  46. ISO 4832:2006; Horizontal Method for the Enumeration of Coliforms. ISO: Geneva, Switzerland, 2006. Available online: https://www.iso.org/standard/38282.html (accessed on 1 November 2022).
  47. ISO 6579:2002; Microbiology of Food and Animal Feeding Stuffs–Horizontal Method for the Detection of Salmonella spp. ISO: Geneva, Switzerland, 2002. Available online: https://www.iso.org/ru/standard/29315.html (accessed on 1 November 2022).
  48. ISO 7932:2004; Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Presumptive Bacillus cereus—Colony-Count Technique at 30 degrees C. ISO: Geneva, Switzerland, 2004. Available online: https://www.iso.org/standard/38219.html (accessed on 1 November 2022).
  49. ISO 13720:2010; Meat and Meat Products—Enumeration of Presumptive Pseudomonas spp. ISO: Geneva, Switzerland, 2010. Available online: https://www.iso.org/ru/standard/45099.html (accessed on 1 November 2022).
  50. ISO 6888-1:2021; Microbiology of the Food Chain—Horizontal Method for the Enumeration of Coagulase-Positive Staphylococci (Staphylococcus aureus and Other Species)—Part 1: Method Using Baird-Parker Agar Medium. ISO: Geneva, Switzerland, 2021. Available online: https://www.iso.org/ru/standard/76672.html (accessed on 1 November 2022).
  51. ISO 11290-1:2017; Microbiology of Food and Animal Feeding Stuffs–Horizontal Method for the Enumeration of Listeria monocytogenes—Part 1, Incorporating Amendment 1. ISO: Geneva, Switzerland, 2017. Available online: https://www.iso.org/standard/60313.html (accessed on 1 November 2022).
  52. Manessis, G.; Kalogianni, A.I.; Lazou, T.; Moschovas, M.; Bossis, I.; Gelasakis, A.I. Plant-Derived Natural Antioxidants in Meat and Meat Products. Antioxidants 2020, 9, 1215. [Google Scholar] [CrossRef]
  53. Liu, H.; Qiu, N.; Ding, H.; Yao, R. Polyphenols contents and antioxidant capacity of 68 Chinese herbals suitable for medical or food uses. Food Res. Int. 2008, 41, 363–370. [Google Scholar] [CrossRef]
  54. Alirezalu, K.; Pateiro, M.; Yaghoubi, M.; Alirezalu, A.; Peighambardoust, S.H.; Lorenzo, J.M. Phytochemical constituents, advanced extraction technologies and techno-functional properties of selected Mediterranean plants for use in meat products. A comprehensive review. Trends Food Sci. Technol. 2020, 100, 292–306. [Google Scholar] [CrossRef]
  55. Awad, A.M.; Kumar, P.; Ismail-Fitry, M.R.; Jusoh, S.; Ab Aziz, M.F.; Sazili, A.Q. Overview of plant extracts as natural preservatives in meat. J. Food Process. Preserv. 2022, 46, e16796. [Google Scholar] [CrossRef]
  56. Mariutti, L.R.; Nogueira, G.C.; Bragagnolo, N. Lipid and cholesterol oxidation in chicken meat are inhibited by sage but not by garlic. J. Food Sci. 2011, 76, C909–C915. [Google Scholar] [CrossRef] [PubMed]
  57. Lorenzo, J.M.; Munekata, P.E.S.; Sant’Ana, A.S.; Carvalho, R.B.; Barba, F.J.; Toldrá, F.; Mora, L.; Trindade, M.A. Main characteristics of peanut skin and its role for the preservation of meat products. Trends Food Sci. Technol. 2018, 77, 1–10. [Google Scholar] [CrossRef]
  58. Ashokkumar, K.; Murugan, M.; Dhanya, M.K.; Pandian, A.; Warkentin, T.D. Phytochemistry and therapeutic potential of black pepper [Piper nigrum (L.)] essential oil and piperine: A review. Clin. Phytosci. 2021, 7, 52. [Google Scholar] [CrossRef]
  59. Feng, Y.; Dunshea, F.R.; Suleria, H.A.R. LC-ESI-QTOF/MS characterization of bioactive compounds from black spices and their potential antioxidant activities. J. Food Sci. Technol. 2020, 57, 4671–4687. [Google Scholar] [CrossRef]
  60. Lee, J.G.; Chae, Y.; Shin, Y.; Kim, Y.-J. Chemical composition and antioxidant capacity of black pepper pericarp. Appl. Biol. Chem. 2020, 63, 35. [Google Scholar] [CrossRef]
  61. Zhang, L.; Lokeshwar, B.L. Medicinal properties of the Jamaican pepper plant Pimenta dioica and Allspice. Curr. Drug Targets 2012, 13, 1900–1906. [Google Scholar] [CrossRef]
  62. Miyajima, Y.; Kikuzaki, H.; Hisamoto, M.; Nikatani, N. Antioxidative polyphenols from berries of Pimenta dioica. Biofactors 2004, 21, 301–303. [Google Scholar] [CrossRef]
  63. Torrijos, R.; Righetti, L.; Cirlini, M.; Calani, L.; Mañes, J.; Meca, G.; Dall’Asta, C. Phytochemical profiling of volatile and bioactive compounds in yellow mustard (Sinapis alba) and oriental mustard (Brassica juncea) seed flour and bran. LWT 2023, 173, 114221. [Google Scholar] [CrossRef]
  64. Martinović, N.; Polak, T.; Poklar Ulrih, N.; Abramovič, H. Mustard Seed: Phenolic Composition and Effects on Lipid Oxidation in Oil, Oil-in-Water Emulsion and Oleogel. Ind. Crop. Prod. 2020, 156, 112851. [Google Scholar] [CrossRef]
  65. Reddy, B.C.; Noor, A.; Sabareesh, V.; Vijayalakshmi, M.A. Preliminary screening of potential flavonoid-subclasses in Myristica fragrans and Cordyline terminalis by LCESI-MS. J. Pharmacogn. Phytochem. 2016, 5, 437–450. [Google Scholar]
  66. Antasionasti, I.; Datu, O.S.; Lestari, U.S.; Abdullah, S.S.; Jayanto, I. Correlation Analysis of Antioxidant Activities with Tannin, Total Flavonoid, and Total Phenolic Contents of Nutmeg (Myristica fragrans Houtt) Fruit Precipitated by Egg white. Borneo J. Pharm. 2021, 4, 301–310. [Google Scholar] [CrossRef]
  67. Gupta, A.D.; Bansal, V.K.; Babu, V.; Maithil, N. Chemistry, antioxidant and antimicrobial potential of nutmeg (Myristica fragrans Houtt). J. Genet. Eng. Biotechnol. 2013, 11, 25–31. [Google Scholar] [CrossRef]
  68. Karna, K.K.; Choi, B.R.; You, J.H.; Shin, Y.S.; Cui, W.S.; Lee, S.W.; Kim, J.H.; Kim, C.Y.; Kim, H.K.; Park, J.K. The ameliorative effect of monotropein, astragalin, and spiraeoside on oxidative stress, endoplasmic reticulum stress, and mitochondrial signaling pathway in varicocelized rats. BMC Complement. Altern. Med. 2019, 19, 333. [Google Scholar] [CrossRef] [PubMed]
  69. Pal, C.B.T.; Jadeja, G.C. Deep eutectic solvent-based extraction of polyphenolic antioxidants from onion (Allium cepa L.) peel. J. Sci. Food Agric. 2019, 99, 1969–1979. [Google Scholar] [CrossRef] [PubMed]
  70. Munir, M.T.; Kheirkhah, H.; Baroutian, S.; Quek, S.Y.; Young, B.R. Subcritical water extraction of bioactive compounds from waste onion skin. J. Clean. Prod. 2018, 183, 487–494. [Google Scholar] [CrossRef]
  71. Campone, L.; Celano, R.; Lisa Piccinelli, A.; Pagano, I.; Carabetta, S.; Sanzo, R.D.; Russo, M.; Ibañez, E.; Cifuentes, A.; Rastrelli, L. Response surface methodology to optimize supercritical carbon dioxide/co-solvent extraction of brown onion skin by-product as source of nutraceutical compounds. Food Chem. 2018, 269, 495–502. [Google Scholar] [CrossRef]
  72. Kwak, J.H.; Seo, J.M.; Kim, N.H.; Arasu, M.V.; Kim, S.; Yoon, M.K.; Kim, S.-J. Variation of quercetin glycoside derivatives in three onion (Allium cepa L.) varieties. Saudi J. Biol. Sci. 2017, 24, 1387–1391. [Google Scholar] [CrossRef]
  73. Kotenkova, E.A.; Kupaeva, N.V. Comparative antioxidant study of onion and garlic waste and bulbs. IOP Conf. Ser. Earth Environ. Sci. 2019, 333, 012031. [Google Scholar] [CrossRef]
  74. Vijayalakshmi, G.; Raja, M.M.; Naik, M.L.; Carbone, V.; Russo, G.L.; Khan, P.S.V. Determination of antioxidant capacity and flavonoid composition of onion (Allium cepa L.) landrace ‘Krishnapuram’ bulb using HPLC-ESI-ITMS. J. Biosci. 2021, 46, 58. [Google Scholar] [CrossRef]
  75. Oku, S.; Aoki, K.; Honjo, M.; Muro, T.; Tsukazaki, H. Temporal Changes in Quercetin Accumulation and Composition in Onion (Allium cepa L.) Bulbs and Leaf Blades. Hortic. J. 2021, 90, 326–333. [Google Scholar] [CrossRef]
Table
Table 1. Recipes for meat pâtés.
Table 1. Recipes for meat pâtés.
IngredientControlE1 #E2 #
Blanched materials, g/kg blanched materials
Beef flank350350350
Beef liver230230230
Lean pork200200200
Pork heart100100100
Wheat flour505050
Powdered cow’s milk202020
Fried onions505050
Auxiliary materials, g/kg unsalted blanched materials
Table salt141414
Sugar333
Ground black pepper111
Ground allspice0.50.50.5
Ground mustard555
Nutmeg0.50.50.5
Auxiliary materials, mL/kg unsalted blanched materials
Beef broth after blanching200132166
OHE *-6834
* OHE—yellow onion (Állium cépa) husk ethanolic extract; # OHE was added instead of broth at volumes of 68 µL/g for experiment 1 (E1) and 34 µL/g for experiment 2 (E2).
Table
Table 2. Proximal analysis of meat pâtés.
Table 2. Proximal analysis of meat pâtés.
Fat, %Protein, %Moisture, %Ash, %Carbohydrates, %
Control9.33 ± 0.0621.27 ± 0.0657.17 ± 0.211.95 ± 0.0410.29 ± 0.12
E18.00 ± 0.20 *28.03 ± 0.15 *53.0 ± 0.17 *2.11 ± 0.06 *8.85 ± 0.43 *
E28.93 ± 0.15 *,#24.0 ± 0.10 *,#55.7 ± 0.20 *,#2.10 ± 0.06 *9.27 ± 0.34 *
* Significant difference as compared to control (based on two-tailed Mann–Whitney U test, p < 0.1); # significant difference between E2 and E1 (based on two-tailed Mann–Whitney U test, p < 0.1).
Table
Table 3. Compounds in meat pâtés and OHE.
Table 3. Compounds in meat pâtés and OHE.
MetaboliteOHE,
µg-eq. Q/100 mL		µg-eq. Q/100 g Meat Pâté
Predicted (Calculated)Control
(C)		Obtained
E1E2E1E2∆ (E1-C)∆ (E2-C)
Delphinidin
3-galactoside		955.5 ± 59.465.0 ± 4.032.5 ± 2.018.8 ± 2.680.4 ± 34.2 *35.7 ± 3.1 *,#61.617.0
Petunidin
3-galactoside		293.5 ± 19.820.0 ± 1.410.0 ± 0.713.1 ± 4.615.6 ± 3.519.1 ± 5.92.56.0
Luteolin-4′-
O-glucoside		804.9 ± 56.953.7 ± 3.927.4 ± 1.930.9 ± 2.261.8 ± 8.2 *43.3 ± 8.9 *30.912.5
Spiraeoside365,200.0
± 0.0		24,833.6
± 0.0		12,416.8
± 0.0		11,041.1
± 158.1		20,809.7
± 372.8 *		16,447.7
± 504.1 *,#		9768.65406.7
Myricitrin6534.4 ± 344.7444.3 ± 23.4222.2 ± 11.7254.9 ± 20.0656.7 ± 8.1 *506.9 ± 32.4 *,#402.1252.0
Isorhamnetin-3-O-beta-D-
Glucoside		2789.6 ± 53.1189.7 ± 3.694.9 ± 1.8180.5 ± 25.8259.0 ± 6.3 *273.4 ± 21.0 *78.492.8
Quercetin 3-O-malonylglucoside441.0 ± 17.530.0 ± 1.215.0 ± 0.60.0 ± 0.015.0 ± 6.4 *8.4 ± 1.7 *15.08.4
Quercetin-3,4′-O-di-beta-
glucoside		1796.9 ± 121.3122.2 ± 8.361.1 ± 4.10.1 ± 0.273.7 ± 3.8 *40.0 ± 9.2 *,#73.539.9
Baimaside449.7 ± 34.030.6 ± 2.315.3 ± 1.20.4 ± 0.819.4 ± 10.2 *11.4 ± 0.3 *19.011.0
Isorhamnetin1831.1 ± 132.9124.5 ± 9.062.3 ± 4.524.2 ± 6.7144.0 ± 7.7 *80.1 ± 11.3 *,#119.855.9
Kaempferol1057.3 ± 29.171.9 ± 2.036.0 ± 1.03.0 ± 1.953.8 ± 10.4 *22.1 ± 1.8 *,#50.819.1
Quercetin174,546.7
± 3596.7		11,869.2
± 244.6		5934.6
± 122.3		77.9
± 10.1		10,279.0 *
± 389.8 *		4563.6 *
± 76.8 *,#		10,201.04485.7
Total556,700.5
± 3495.1		37,855.6
± 237.8		18,927.8
± 118.8		11,644.9
± 164.3		32,468.2 *
± 664.1 *		21,949.4
± 417.5 *,#		20,823.310,304.5
* Significant difference as compared to control (based on two-tailed Mann–Whitney U test, p < 0.1); # significant difference between E2 and E1 (based on two-tailed Mann–Whitney U test, p < 0.1).
Table
Table 4. TAC of meat pâtés during storage at 4 °C.
Table 4. TAC of meat pâtés during storage at 4 °C.
DayTACFRAP, µmol-equiv. Q/ 100 g Meat Pâté
Control (C)E1E2∆ (E1-C)∆ (E2-C)
032.83 ± 1.1972.94 ± 1.85 *50.52 ± 1.27 *,#40.1117.69
329.96 ± 0.7973.35 ± 0.30 *49.88 ± 0.67 *,#43.3919.92
526.19 ± 1.2164.25 ± 1.03 *42.97 ± 0.20 *,#38.0616.78
725.06 ± 0.2058.42 ± 0.29 *37.73 ± 0.29 *,#33.3612.67
1421.69 ± 0.4450.13 ± 0.97 *36.59 ± 0.14 *,#28.4414.90
* Significant difference as compared to control (based on two-tailed Mann–Whitney U test, p < 0.1); # significant difference between E2 and E1 (based on two-tailed Mann–Whitney U test, p < 0.1).
Table
Table 5. Decreases in TACFRAP during meat product processing.
Table 5. Decreases in TACFRAP during meat product processing.
µmol-equiv. Q/100 g Meat PâtéDecrease in TACFRAP in E1, %µmol-equiv. Q/100 g Meat PâtéDecrease in TACFRAP in E2, %
Amount in Recipe E1∆ (E1-C)Amount in Recipe E2∆ (E2-C)
44.436.67 ± 5.8617.4122.216.39 ± 2.759.45
Table
Table 6. PV of meat pâtés during storage at 4 °C.
Table 6. PV of meat pâtés during storage at 4 °C.
DayPV, mmol. Active O2/kg of Fat
ControlE1E2
02.07 ± 0.342.09 ± 0.161.35 ± 0.25 *,#
32.0 ± 0.372.44 ± 0.12 *2.57 ± 0.14 *
72.83 ± 0.042.98 ± 0.142.61 ± 0.05 *,#
103.62 ± 0.093.32 ± 0.03 *3.54 ± 0.06 #
143.65 ± 0.163.84 ± 0.224.07 ± 0.02 *
284.19 ± 0.113.92 ± 0.233.96 ± 0.23
p-value 1
(0–28 days)		0.0140.0120.014
p-value 1
(0–7 days)		0.0970.0500.097
* Significant difference as compared to control (based on two-tailed Mann–Whitney U test, p < 0.1); # significant difference between E2 and E1 (based on two-tailed Mann–Whitney U test, p < 0.1); 1 based on a Freidman ANOVA, p < 0.05.
Table
Table 7. pH of meat pâtés during storage at 4 °C.
Table 7. pH of meat pâtés during storage at 4 °C.
DaypH Value
ControlE1E2
06.07 ± 0.066.09 ± 0.045.98 ± 0.05 #
35.95 ± 0.035.92 ± 0.036.05 ± 0.04 *,#
75.95 ± 0.025.83 ± 0.03 *6.02 ± 0.04 *,#
105.89 ± 0.035.83 ± 0.02 *6.00 ± 0.01 *,#
145.93 ± 0.045.87 ± 0.02 *6.02 ± 0.01 *,#
286.0 ± 0.055.91 ± 0.02 *6.06 ± 0.03 #
p-value 1
(0–28 days)		0.0250.0140.187
p-value 1
(0–7 days)		0.0970.0500.202
* Significant difference as compared to control (based on two-tailed Mann–Whitney U test, p < 0.1); # significant difference between E2 and E1 (based on two-tailed Mann–Whitney U test, p < 0.1); 1 based on a Freidman ANOVA, p < 0.05.
Table
Table 8. TBARS of meat pâtés during storage at 4 °C.
Table 8. TBARS of meat pâtés during storage at 4 °C.
DayTBARS, µmol/100 g Meat Pâté
ControlE1E2
02.27 ± 0.301.96 ± 0.191.94 ± 0.36
34.57 ± 0.321.89 ± 0.33 *2.20 ± 0.35 *
75.83 ± 3.832.11 ± 0.13 *2.96 ± 0.73 *
106.07 ± 0.172.80 ± 0.57 *2.26 ± 0.34 *
144.79 ± 0.732.29 ± 0.69 *2.83 ± 0.53 *
p-value 1
(0–14 days)		0.0090.1140.027
* Significant difference as compared to control (based on two-tailed Mann–Whitney U test, p < 0.1); 1 based on a Freidman ANOVA, p < 0.05.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Chernukha, I.; Kupaeva, N.; Khvostov, D.; Bogdanova, Y.; Smirnova, J.; Kotenkova, E. Assessment of Antioxidant Stability of Meat Pâté with Allium cepa Husk Extract. Antioxidants 2023, 12, 1103. https://doi.org/10.3390/antiox12051103

AMA Style

Chernukha I, Kupaeva N, Khvostov D, Bogdanova Y, Smirnova J, Kotenkova E. Assessment of Antioxidant Stability of Meat Pâté with Allium cepa Husk Extract. Antioxidants. 2023; 12(5):1103. https://doi.org/10.3390/antiox12051103

Chicago/Turabian Style

Chernukha, Irina, Nadezhda Kupaeva, Daniil Khvostov, Yuliya Bogdanova, Jutta Smirnova, and Elena Kotenkova. 2023. "Assessment of Antioxidant Stability of Meat Pâté with Allium cepa Husk Extract" Antioxidants 12, no. 5: 1103. https://doi.org/10.3390/antiox12051103

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop