1. Introduction
Currently, agro-food by-product re-valorization is important for reducing generated wastes and could be achieved by implementing a circular economic model to protect the environment. Moreover, these by-products can be used as an alternative source of natural additives to improve the quality of food products [
1].
Mayonnaise is one of the most popular and well-liked condiments used in a variety of foods to improve flavor and taste. The ingredients used for mayonnaise formulation include egg, either as a whole or egg yolk, vinegar, water, mustard, spices, and vegetable oil, such as soybean, rapeseed, sunflower, or corn oil [
2]. Mayonnaise preparation uses different emulsifying proportions of vegetable oil (65–80%) with other ingredients, such as eggs and mustard [
3]. Although it is highly appreciated for its flavor and taste, mayonnaise is susceptible to lipid oxidation due to the high polyunsaturated fatty acid content (PUFA) from vegetable oil [
4]. The lipid oxidation process is initiated at the interface between oil and aqueous phases and continues after the oil phase, during product storage. Lipid oxidation affects the organoleptic and nutritional properties and even the storage stability of mayonnaise [
5,
6]. Factors such as fatty acid profile, pro-oxidants, and storage conditions make multiphase food systems, such as mayonnaise, susceptible to oxidation [
5,
7]. It is well established that lipid oxidation degrades product quality through rancid odors production, and unpleasant flavors, and potentially compromises food safety through the production of toxic chemicals [
8]. Synthetic ingredients are commonly used to prevent lipid degradation in mayonnaise composition, which is included in the recipe to accomplish a variety of functions to ensure product quality. Synthetic antioxidants, such as butylated hydroxyanisole (BHA), ethylenediaminetetraacetic acid (EDTA) and butylated hydroxytoluene (BHT), are added to mayonnaise composition to control the oxidative degradation and avoid fast degradation [
7,
9]. However, there is increased concern about food products that include synthetic ingredients, that may seriously affect consumers’ health. Although synthetic antioxidants are inexpensive and effective, they are not widely accepted by consumers. Therefore, the food industry is currently focused on substituting natural antioxidants with those obtained from natural resources [
10].
Plant materials represent rich sources of phenolic compounds which demonstrate a broad range of biological functions, including antioxidative protection, antimicrobial, antimutagenic, and anti-inflammatory properties [
11]. Therefore, the food industry has been focused on using natural sources of bioactive compounds with multiple functionalities to obtain healthy products. Numerous studies have demonstrated that antioxidants derived naturally from phenolic-rich plant components, such as extracts from berries [
12], olives [
13], olive leaves [
14], grape seed [
15], buckwheat hull [
10], purple corn [
6], or sesame sprouts powder [
16], can increase the stability and delay the oxidation process of emulsions.
Beetroot (
Beta vulgaris L.) belongs to the
Amaranthaceae family. It is one of the most potent vegetables concerning antioxidant activity, mainly because it is rich in betalains and phenolic compounds [
17]. Betalains are water-soluble pigments that have been authorized by the European Union as E-162 for use in food as colorants [
18]. Moreover, beetroot is an important source of beneficial compounds to health, including: folate, which offers protection against congenital disabilities; iron, which helps prevent and treat anemia; and dietary fibers, with a crucial role in improving colon health [
19]. Although the resulting pomace from the beetroot processing is abundant in dietary fibers, antioxidants, and minerals, it is not fully valorized and might be targeted as a significant source of naturally occurring antioxidants that can help enhance food quality. Therefore, incorporating beetroot by-products into food products such as mayonnaise may be affordable. Beetroot has been used as a natural ingredient in different food products for obtaining value-added products. Hence in the reviewed literature beetroot added products were identified that improved both nutritional and sensorial properties, such as yogurt with beetroot juice [
20], yogurt beetroot powder [
21], ice cream with beetroot puree [
22], and with beetroot juice [
23], and cakes enriched with beetroot powder [
24]. In this way, beetroot by-products can be used as low-cost food components, hence reducing food waste. Therefore, plant wastes could represent a nutraceutical source used in functional food production [
25].
A previous study has shown that either raw or processed beetroot may be successfully incorporated in the mayonnaise composition to replace synthetic antioxidants and extend shelf-life [
4]. The present research focused on utilizing freeze-dried beetroot peels as a source of natural antioxidants and other lipophilic bioactive compounds into the mayonnaise composition to obtain a value-added product. Furthermore, the impact of beetroot peels powder (BPP) supplementation on the phytochemical composition, sensory characteristics, viscosity, color, and textural properties of the mayonnaise were also investigated.
2. Materials and Methods
2.1. Materials
Beetroot was collected from a Galați local farmer. All ingredients used for the mayonnaise were obtained at a local market, including refined sunflower oil, vinegar, egg powder, lemon salt, and salt. Folin–Ciocalteau reagent, sodium carbonate, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 6-hydroxi-2,5,7,8-tetrametilcroman-2-carboxilic acid (Trolox), ethanol (70%), and citric acid were supplied from Sigma-Aldrich (Steinheim, Germany).
2.2. Beetroot Peels Powder (BPP) Preparation
The beetroots were washed and dried. Then, the peel was removed using a knife, and the collected peels were lyophilized using Alpha 1–4 LD plus equipment (Christ, Germany), at −42 °C, under a pressure of 0.10 mBar, for 48 h. Following that, the freeze-dried peels were ground to a fine powder using a grinder MC 12 (Producer Stephan, Germany) and stored in the dark at room temperature.
2.3. Extraction of Polyphenols and Betalains from Beetroot Peels Powder and Antioxidant Activity Evaluation
An amount of 1 g of BPP was added to 9 mL of a 50% (v/v) ethanol solution and 1 mL citric acid (1%) and then manually mixed for 1 min. Samples were sonicated in a water bath (Smart Sonic cleaner MRC) for 50 min, at 40 °C. After that, the extract was centrifuged at 5000 rpm for 15 min, at 20 °C, followed by collection of the supernatant. The residue was extracted again with a solvent (10- and 20-mL ethanol 50%), and the extracts were mixed and kept at 4 °C.
2.3.1. Betalains
The concentration of betalains in the BPP extract was determined using the spectrophotometric method described by Stintzing et al. [
26]. The absorbances of the betalains were read at 538 nm for betacyanins and 480 nm for betaxanthins. The betalain concentration (BC) was determined as follows:
where A is the sample absorbance, DF—the dilution factor, l—cuvette pathlength (1 cm), W d/g is the amount of BPP, and Vd is the volume of BPP solution. The molecular weights (MW) and molar extinction coefficients of betacyanins and betaxanthins were used to quantify them (e) (MW = 550 g/mol; e = 60.000 L/mol cm in H
2O) and (MW = 308 g/mol; e = 48,000 L/mol cm in H
2O) were applied.
2.3.2. Total Polyphenolic Content
The total soluble phenolics contained in BPP extract were quantified using the Folin–Ciocalteu assay in accordance with the procedure specified by Turturica et al. [
27]. Briefly, to tubes containing 15.8 mL distilled water, 1.0 mL Folin–Ciocalteau solution, and 0.2 mL BPP extract was added. After 10 min, 3 mL of Na
2CO
3 20% was added to the mixture. The mixture obtained was stored for 60 min at room temperature in the dark, prior to determining the absorbance at 765 nm against a control (pure ethanol). Milligrams of gallic acid equivalents per gram of dry weight (mg GAE/g DW) were used to express the results.
2.3.3. Antioxidant Activity
The DPPH technique was used to determine the antioxidant activity, and the findings were represented as Trolox equivalents in millimoles per gram of dry weight (mM TE/g DW) [
28]. Briefly, the absorbance of the blank, 3.9 mL DPPH solution 0.1 M in ethanol, was measured at 515 nm. Then, to the reaction mixture of 3.9 mL DPPH solution 0.1 M, a volume of 0.1 mL of BPP extract was added, and afterward, the mixture was kept for 1 h and 30 min at room temperature in the dark before the absorbance at
λ = 515 nm was recorded.
2.4. Mayonnaise Preparation
A mayonnaise recipe was created by combining the following ingredients in the following weight ratios (w/w): sunflower oil (80%), egg powder (8%), water (7%), vinegar (2%), lemon salt (3%), salt (0.3%) and different proportions (1.5%, 3%, 5%, and 7%) of BPP hydrated with water (1:1).
To begin, a coarse emulsion was created in water by dissolving egg powder, lemon salt, salt, and vinegar. Mayonnaise was prepared by gradually adding the oil to the aqueous mixture and quickly blending the components with a Morphy Richards hand blender1.5 (Argos, Milton Keynes, UK) for 10–15 min. The BPP was further added to the mayonnaise at four different concentrations of 1.5%, 3%, 5%, and 7%, while continuing mixing until the purple samples were uniform and coded S1, S2, S3, and S4, respectively. The control mayonnaise was produced in the same manner as the experimental mayonnaise but without the addition of BPP. The mayonnaise samples obtained were kept at 4 °C until measurements were taken.
2.5. Characterization of Phytochemicals, Physicochemical, and Antioxidant Activity of Mayonnaise Supplemented with BPP
The physicochemical parameters of mayonnaise samples (e.g., moisture content, protein content, fat content, ash content, total sugar content, and energy value) were estimated using AOAC methods [
29]. The betalains, phenolic content, and antioxidant activity of mayonnaise enhanced with BPP, were assessed using the techniques described above in
Section 2.3.1,
Section 2.3.2 and
Section 2.3.3.
2.6. Color Measurements
Color attributes of the supplemented mayonnaise containing varying amounts of BPP were quantified using the CR300 Chroma Meter (Konica Minolta, Sensing Americas, Inc., Ramsey, NJ, USA) provided with a D65 Illuminant. The CIELAB color parameters (L*, a*, and b*) were obtained in triplicate following equipment calibration against the white plate. The total color change (ΔE) was also calculated for each sample according to Polat et al. [
30] using the formula below:
where the abbreviation of color parameters represents m—sample without BPP, and bm—sample with BPP.
2.7. Textural Profile Analysis
The texture profile analysis (TPA) approach was used to examine the textural features of the mayonnaise samples, using a CT3-1000 Texture Analyzer (Brookfield Ametek, Chandler, AZ, USA). Double dispersion into a 38.1 mm diameter acrylic cylinder sample was used to achieve a depth of 20 mm. The test speed was set to one millisecond per second, the trigger load to 0.067 N, and the load cell to 9.8 N. The textural parameters firmness, cohesiveness, springiness, adhesiveness, and chewiness were calculated using the TexturePro CT V1.5 software. Each sample was subjected to three determinations. The samples were maintained at room temperature for two hours before testing.
2.8. Viscosity
The viscosity of the mayonnaise samples was assessed using a rotational viscometer (Brookfield Viscometers Ltd., Harlow, UK), equipped with an LV2 Spindler, with a diameter of 18.72 mm and a height of 115 mm. The mayonnaise samples were added into cylindrical plastic containers, with 37 mm diameter and 200 mm height. All measurements were performed at room temperature. The dynamic viscosity was read directly for different values of the rotational speed of the spindler. A thixotropic loop measurement was performed first, increasing the shear rate from 0.06 to 21.2 s−1 followed by decreasing it back to 0.06 s−1 and reading the values of dynamic viscosity. The power-law model was used to express the dependence between shear rate and shear stress.
2.9. Oxidative Stability
All samples were kept in glass containers and stored in the dark at a refrigerated temperature of 4 °C for 28 days. To evaluate the samples’ oxidative stability, titratable acidity, peroxide value, phytochemical and color parameters were monitored. Titratable acidity was assessed using the official method by AOAC [
29]. The results were reported as mg KOH per gram of sample (mg KOH/g). Peroxide value was quantified using the method described by AOAC [
29]. Extraction of oils from the mayonnaise samples before the peroxide value determination was carried out in accordance with the procedure specified by Park et al. [
10]. In brief, the samples were freeze-dried at −20 °C for 24 h and then kept at room temperature for 2 h in the dark for breaking the emulsion. Then, the samples were centrifuged at 1000 rpm for 10 min, and the oily phase was used for peroxide value analysis. Peroxide value was reported as milliequivalents per kg of the sample (meq/kg).
2.10. Sensory Analysis
The sensory attributes of mayonnaise samples were evaluated by 20 untrained consumers ranging from 25 to 60 years old (80% women and 20% men). A training session was provided to the panelists before sensory analysis. A 9-point hedonic assessment with scores from 1 (very dislike) to 9 (very like) was used in the questionnaire to evaluate the mayonnaise samples. Spreadability, consistency, texture, and color uniformity were assessed for appearance and structure; aroma, odor, and taste were reviewed for flavor and perception; aftertaste and acceptability for overall acceptance were also rated [
7,
14]. The samples were served at room temperature, and water and crackers were served to rinse and eat between tasting samples. The data were adjusted to the average score assigned to each attribute by each panelist.
2.11. Statistical Data
All analyses were conducted in triplicate, and the data are presented as mean and standard deviation. Significant differences among results were identified using analysis of variance (ANOVA). Tukey’s test was applied using Minitab 18 software to determine which pairwise comparisons were significant. For all tests, p-values of p < 0.05 were considered statistically significant.
4. Conclusions
The current paper proposes a unique strategy for valorizing beetroot by-products as a source of bioactive chemicals to develop novel value-added products. BPP extract characterization revealed a high concentration of phenolic compounds and exhibited significant antioxidant activity. BPP functionality was further evaluated by adding it to a mayonnaise composition. The mayonnaise samples’ antioxidant activity increased with BPP content and, due to the enhanced polyphenol content, demonstrated a greater nutritional quality than the control sample. Due to the presence of considerable levels of beetroot pigments, BPP addition resulted in a rise in the redness value and a reduction in the yellowness of the mayonnaise sample. Moreover, the addition of BPP in mayonnaise samples determined higher firmness and an improvement of adhesion and chewiness, giving the product a soft texture. The viscosity of mayonnaise was also significantly improved. Sensory evaluation of value-added mayonnaise indicated that the addition of BPP enhanced the product’s color attributes and caused no noticeable impact on the odor, aftertaste, spreadability, and overall acceptance score of the samples. Moreover, using by-products obtained from the industrial processing of beetroots, by considering them a source of biologically active compounds, may become a viable alternative to synthetic dyes, flavorings, and antioxidants. This can have multiple uses in the food industry and can contribute to the reduction of wastes and support the implementation of a circular economic model for the protection of the environment.