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The Effect of Microencapsulation of Phenolic Compounds from Lemon Waste by Persian and Basil Seed Gums on the Chemical and Microbiological Properties of Mayonnaise
1Department of Food Science and Technology, Islamic Azad University of Varamin-Pishva Branch, Varamin, Tehran 33317-74895, Iran
2Department of Food Science and Technology, Islamic Azad University, Isfahan (Khorasgan) Branch, Isfahan 81595-158, Iran
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Prev Nutr Food Sci 2021; 26(1): 82-91
Published March 31, 2021 https://doi.org/10.3746/pnf.2021.26.1.82
Copyright © The Korean Society of Food Science and Nutrition.
Abstract
Keywords
INTRODUCTION
Mayonnaise is a widely consumed food product used as seasoning for different types of food (Liu et al., 2007). Due to its structure and composition, mayonnaise is highly susceptible to chemical and microbial deterioration. Chemical deterioration includes oxidation and hydrolysis of fat and oil types, which increases the content of compounds like peroxide, leading to sourness and changes to its color, taste, and texture (Frankel et al., 2002). Mayonnaise is susceptible to deterioration due to the structure of its lipid molecules, and their interactions with other molecules in their immediate vicinity, greatly influencing susceptibility to lipid oxidation (Coupland and McClements, 1996). Mayonnaise oxidation is intensified by free radicals generated from fat oxidation with other molecules like proteins, carbohydrates, and vitamins in the emulsion. In addition, undesirable changes in pH and acidity caused by microbial or chemical activity in mayonnaise leads to microbial deterioration (Shahidi and Zhong, 2005).
Therefore, synthetic antioxidants and chemical conservatives are added to mayonnaise to control oxidation and microbial activity, which has economic advantages despite the compounds being harmful to human health (Kwon et al., 2015). For example, the food conservative sodium benzoate inhibits activity of bacteria and fungi in acidic environments (e.g., in sauces and salad seasonings). However, sodium benzoate damages cell mitochondrial DNA, consequently inactivating the cell and leading to development of the Alzheimer’s and Parkinson’s diseases (Daescu et al., 1986).
Consumption of food prepared with natural sources of phenolic compounds with a wide range of antioxidant, antimicrobial, and anti-mutagenic activities has recently increased (Kong et al., 2003). In Iran, 650,000 tons of citrus are produced annually, which accounts for 9% of the global production (Vand and Abdullah, 2012).
Lemon is a species of citrus. The large amounts of lemon waste (peels, seeds, and pulps) produced by lemon juice industries have attracted attention. Lemon exhibits antimicrobial and antioxidant activity as it contains phenolic compounds, including flavonoids, coumarin, and limonene (Altunkaya et al., 2013).
Polyphenols contain a large group of bioactive compounds, and are present in fruit, vegetables, seeds, and other parts of plants. For humans, diet is one of the most important sources of polyphenols, which exhibit antioxidant properties and roles in food preservation and human health (Shariatifar et al., 2014). These compounds are sensitive to heat, light, and oxidation due to unsaturated bands within their molecular structures (Halliwell, 2008).
The strong antioxidant activity of phenolic compounds arises from their ability to scavenge free radicals. Indeed, radical scavenging activity is attributed to the presence of hydroxyl groups that replace aromatic rings. Phenolic compounds are present in many plant-based foods that have antioxidant activity (Lakshmanashetty et al., 2010).
Destruction of phenolic compounds can be prevented by microencapsulation, a process whereby bioactive compounds are trapped by biolipids, protecting them against the environment, thus increasing stability (Belščak-Cvitanović et al., 2011). Encapsulation can be achieved by biopolymers like starch compounds, agar, alginate, chitosan, and other gums (King, 1995). Arabic gum (AB) extracted from
Persian gum (PG) is composed of galactose and arabinose and is applied as an emulsifier in acidic pH emulsion systems, high salt foods, and thermal processes. Similar to other gums, PG is water soluble and produces a sticky and viscous solution (Golkar et al., 2018).
Basil seed gum (BSG) exhibits a heterosaccharide structure that includes glucomannan, glucan, and xylan. BSG is an exclusive hydrocolloid categorized as an anionic gum and is added as an active ingredient in food formulation to increase consistency and gelation and to control microstructure, texture and aroma during storage (Dickinson, 2003; Hosseini-Parvar et al., 2010).
In the present study, we extracted phenolic compounds from lemon waste (peels and edible pulps) and microencapsulated them in gel matrixes of PG and BSG as substitutes to synthetic antioxidants and sodium benzoate in mayonnaise production. We assessed the effects of encapsulation on physical, chemical, sensory, and microbial properties of mayonnaise over one month. These phenolic compounds may represent new preservatives and flavorings for use in mayonnaise formulation.
MATERIALS AND METHODS
Materials
The primary ingredients consumed during mayonnaise preparation (oil without antioxidants, vinegar, mustard, pasteurized eggs, sugar, and salt) were purchased from a local supermarket in Isfahan, Iran. Lemons were obtained from a local farm in Shiraz. AB was provided from Sigma-Aldrich Co. (St. Louis, MO, USA). BSG and PG were purchased from a local market in Isfahan, Iran. Folin-Ciocalteau’s phenol reagent and 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) were purchased from Merck KGaA (Darmstadt, Germany) and Sigma-Aldrich Co., respectively. Methanol, hexane, potassium chloride, trichloroacetic acid, and sodium bicarbonate were purchased from Sigma-Aldrich Co.. All other chemicals or solvents used were of analytical grade.
Extraction of polyphenols from lemon waste
Polyphenols were extracted from lemon by incubating 16 g lemon with 100 mg of 70% ethanol in an ultrasonic bath (60 Hz frequency, 60°C) for 2 h. The extracts were removed by a vacuum rotary evaporator machine at 40°C. The yield extract was filtered and then incubated in a freeze dryer for 48 h (Inoue et al., 2010).
Preparation the PG and BSG
Extraction of mucilage from Basil seeds was achieved following the method described by Hosseini-Parvar et al. (2010) with slight modifications. Extraction was performed using a water/seed ratio of 50:1 at 65°C, pH 7, for 20 min whilst stirring (500 rpm, RH basic 2, IKA, Staufen, Germany). A sterile extractor equipped with a rotating rough plate was used to separate the mucilage from the swollen seeds. The mucilage was then centrifuged and filtered to remove insoluble impurities. Finally, the filtrate was freeze-dried (Dena Vacuum, Tehran, Iran) at −40°C for 24 h and stored under dry, cool, and sterile conditions.
PG was prepared according to methods described in previous studies (Fadavi et al., 2014; Golkar et al., 2015a; Golkar et al., 2015b). Briefly, impurities were removed from lighter colored PG samples and the samples were milled (AQC 109, Agromatic AG, Zürich, Switzerland) and sieved (<500 μm). PG powder was then dissolved in distilled water with stirring (23°C, 500 rpm, RH basic 2, IKA) for 2 h and stored in a refrigerator for 24 h. The solution was centrifuged at 9,000 rpm for 15 min (PIT 320R, Hettichlab, Düsseldorf, Germany) to remove insoluble residues and the supernatant was collected and dried at −40 ºC for 24 h.
Preparation of microcapsules
PG and BSG were encapsulated using polyphenol compounds as coating agents following the method described by Rutz et al. (2013) with some modifications. To prepare the microcapsules, mixtures of hydrocolloids at different PG/BSG ratios (1:0, 1:1, and 0:1) with 15% (w/w) total biopolymer were dissolved in water. Polyphenol extracts (containing 0.17 g of polyphenol) were then added so that the ratio of polyphenol extracts to wall material was 1:3. The mixture was stirred (500 rpm, RH basic 2, IKA) for 12 h at room temperature (23°C) and freeze dried (at −40°C for 24 h).
Evaluation the of microcapsule properties
Preparation of mayonnaise
Mayonnaise was produced from the oil without antioxidant (65.2%), pasteurized egg (13.85%), water (8.2%), vinegar (7.7%), salt (1.5%), and mustard (0.3%) (all amounts are stated as % w/w). First, eggs and the powdered materials (salt, sugar, mustard powder, and citric acid) were homogenized, and then oil was added until a mayonnaise emulsion had been formed. Next, vinegar was added to the mayonnaise emulsion, followed by additives (PG, BSG, or PG/BSG extracts; 1,000 ppm in all samples) and distilled water whilst mixing.
Evaluation of the mayonnaise properties
where Asample is absorbance of sample, Ablank is absorbance of reagent blank, and m is the amount of oil sample amount (mg).
Statistical analysis
Statistical analysis was performed based on completely randomized factorial design and analysis of variance (ANOVA) using SAS 9.1 software (SAS Institute Inc., Cary, NC, USA). Mean comparison tests were performed using least significant difference (LSD) methods at 5% significant level.
RESULTS AND DISCUSSION
Assessment of microcapsule properties
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Table 1 . Total phenolic content, antioxidant activity, encapsulation efficiency, WSI and WAI of lemon waste extracts and microcapsules during storage
Samples Total phenolic content (mg gallic acid/g) Antioxidant activity (%) Encapsulation efficiency (%) WSI (g/100 g dry solid) WAI (g/100 g dry solid) BSG 16.87±0.25b 61.19±0.34b 70.72±0.76a 11.53±0.69a 24.76±0.65c PG/BSG 17.91±0.36b 58.23±0.81c 67.07±0.46b 8.33±0.27b 32.51±0.60a PG 13.92±0.33c 53.95±0.38d 65.06±0.40b 13.28±0.59a 27.11±0.03b Extract 30.50±0.25a 73.21±0.23a ? ? ? The results were expressed as mean±standard deviation of three determinations.
Different letters (a-d) in the column indicate significant differences (
P <0.05).BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated; WSI, water solubility index; WAI, water absorption index.
Many studies have shown a direct correlation between phenolic content and antioxidant properties, whereby PG/BSG samples with the highest phenolic compound contents have lower antioxidant properties compared with BSG samples. The antioxidant properties of polyphenols arise from their redox properties, which allow them to act as reducing agents, hydrogen donators, metal chelators, and single oxygen quenchers (Piluzza and Bullitta, 2011).
The antioxidant activity of lemon extracts was higher than that of the encapsulated samples. This may be attributed to deficient extraction of the phenolic compounds in microcapsules and extract reaction with the capsule membrane, leading to reduction in antioxidant activity. da Rosa et al. (2014) obtained similar results, indicating extract and microcapsule inhibition of 93.07% and 80.38∼90.75%, respectively.
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Figure 1. SEM micrographs of encapsulated lemon waste extracts with PG (A), BSG (B), and PG/BSG (C). BSG, basil seed gum; PG, Persian gum.
In general, freeze-dried encapsulated powder had an irregular shape and flaky structure. Furthermore, micro grooves may be formed due to ice sublimation during freeze-drying. In a previous study, Pasrija et al. (2015) reported similar morphologies for freeze-dried microcapsules containing green tea polyphenols. SEM images of BSG showed that BSG has a fibril structure and contains a globular structure similar to that of the scattered cotton (Naji-Tabasi and Razavi, 2017).
Samples coated with PG/BSG showed the highest WAI (32.51±0.60%), whereas lowest WAI were observed for those coated with PG (24.76±0.65%). Therefore, samples coated with PG is less likely to be influenced by humidity and is more stable under storage conditions (Tan et al., 2015). These results suggest that the percentage of water absorption decreases with increased water solubility of the microcapsules. In a previous study, Moreira et al. (2009) obtained similar results for acerola pomace extracts. Furthermore, according to Ahmed et al. (2010) agglomeration of the encapsulated particles could contribute to a smaller WAI and, subsequently, increase WSI.
Assessing the mayonnaise properties
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Table 2 . Acid values of mayonnaise samples prepared using extracts and microcapsules of lemon waste during storage (unit: mg KOH/g)
Samples 0 10 20 30 Control 0.84±0.016abD 0.87±0.024aC 0.93±0.004aB 1.04±0.005aA BSG 0.83±0.003abB 0.81±0.006bC 0.83±0.003dB 0.87±0.003eA PG/BSG 0.85±0.004aC 0.89±0.003aB 0.90±0.005bA 0.90±0.001dA PG 0.82±0.006bD 0.90±0.006aB 0.89±0.001cC 0.91±0.003cA Extract 0.84±0.016abD 0.87±0.024aC 0.90±0.004bB 0.95±0.003bA The results were expressed as mean±standard deviation of three determinations.
Different small letters (a-e) in the same column and capital letters (A-D) in the same row indicate significant (
P <0.05) differences during the 30-day storage.Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
After 30 days of storage, POV peaked at 27.32±0.1 meqO2/kg oil in the control samples, with significant differences (
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Figure 2. Peroxide values (meqO2/kg) measured as peroxide and hydroperoxide concentrations in lipid extracts of mayonnaise samples prepared using extracts and microcapsules of lemon waste with PG and BSG during storage. Different letters (a-l) indicate significant (
P <0.05) differences between results. Values are mean±standard deviation of three experimental repeats. Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
Evidence of encapsulation improving bioactivity and bioavailability of polyphenols has been reported in previous studies (Fang and Bhandari, 2010). However, it has also been reported that higher concentrations of extracts may act as better pro-oxidants (Sarah et al., 2010). Rasmy et al. (2012) assessed the antioxidant effect of rosemary alcoholic extracts on mayonnaise formulation and found that applying 400 μg of extract (concentration in 1 g of mayonnaise) reduced POV compared with the control group. Moreover, similar to the increase in POVs after 4 months, mayonnaise-containing rosemary followed a slower increase compared to butylated hydroxyanisole antioxidants.
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Figure 3. Thiobarbituric acid (TBA) [mg malondialdehyde (MA)/kg] reactive substance value of lipid extracts of mayonnaise samples prepared using extracts and microcapsules of lemon waste with PG and BSG during storage. Different letters (a-j) indicate significant (
P <0.05) differences between results. Values are mean±standard deviation of three experimental repeats. Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
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Table 3 . Color measurements of mayonnaise samples prepared using extracts and microcapsules of lemon waste during storage
Samples L* a* b* Control 91.77±0.24a ?1.31±0.03a 8.48±0.07b BSG 85.30±0.10d ?1.73±0.02c 8.51±0.13b PG/BSG 87.02±0.29c ?1.68±0.06bc 9.39±0.15ab PG 89.31±0.13b ?1.58±0.03b 10.21±0.06a Extract 91.63±0.20a ?1.51±0.01b 10.35±0.10a The results were expressed as mean±standard deviation of three determinations.
Different letters (a-d) in the column indicate significant differences (
P <0.05).Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
There were no significantly differences in the L* values of the control samples and extract samples. The lowest L* values were observed for BSG sample (85.30±0.10). Furthermore, the samples containing microcapsules had lower transparency compared with the control samples. The highest a* values (red-green) were determined for the control samples (−1.31±0.03), with the lowest values recorded for the BSG samples (−1.73±0.2). In addition, the lowest b* (green-blue) value was observed for the control sample.
Adding microcapsules to prepared mayonnaise samples decreased the L* and a* indexes, and increased the b* indexes. The reduction in the L* index is due to increases in particle diameter, which reduces light differentiation and product lightness. In a previous study, McClements and Demetriades (1998) reported that mayonnaise transparency decreases as the diameters of particles diameter in the emulsion increase.
Negative a* values were reported in the mayonnaise samples. This change in color toward greenness may be due to formation of phenolic polymer compounds. Adding PG increased yellowness (b*), whereas basil mucilage induced the opposite effect due to the color pigments in the extracts and gums. Our results are in line with those of a study by Amiri Aghdaei et al. (2014) which showed decreases in transparency (L*) and yellowness (b*) by addition of
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Figure 4. Total bacterial counts of mayonnaise samples prepared using extracts and microcapsules of lemon waste with PG and BSG during storage. Different letters (a-k) indicate significant (
P <0.05) differences between results. Values are mean±standard deviation of three experimental repeats. Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated; CFU, colony-forming unit.
In all samples, the total microorganism count was <103, resulting from organic acids in the mayonnaise reducing pH and acting as an antimicrobial material (Ma and Boye, 2013). All the samples indicated negative responses to
In the current study, microencapsulation of phenolic compounds extracted from the lemon waste was performed by PG, BSG, and BSG/PG. Highest EE and antioxidant activity were observed for sample coated with BSG (70.72% and 61.19%, respectively). Furthermore, we observed increased chemical stability of microencapsulated phenolic compounds. Samples containing microcapsules reduced peroxide production, stabilized free fatty acids, and prevented increases in acidity by eliminating and preventing formation of free radicals in the mayonnaise. Mayonnaise samples prepared using PG/BSG had significantly (
ACKNOWLEDGEMENTS
The authors would like to acknowledge the Islamic Azad University (IAU), Khorasgan (Isfahan) Branch, for their valuable help, and cooperation with this project.
AUTHOR DISCLOSURE STATEMENT
The authors declare no conflict of interest.
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Article
Original
Prev Nutr Food Sci 2021; 26(1): 82-91
Published online March 31, 2021 https://doi.org/10.3746/pnf.2021.26.1.82
Copyright © The Korean Society of Food Science and Nutrition.
The Effect of Microencapsulation of Phenolic Compounds from Lemon Waste by Persian and Basil Seed Gums on the Chemical and Microbiological Properties of Mayonnaise
Shima Shaygannia1 , Mohammad Reza Eshaghi1
, Mohammad Fazel2, and Mahnaz Hashemiravan1
1Department of Food Science and Technology, Islamic Azad University of Varamin-Pishva Branch, Varamin, Tehran 33317-74895, Iran
2Department of Food Science and Technology, Islamic Azad University, Isfahan (Khorasgan) Branch, Isfahan 81595-158, Iran
Correspondence to:Mohammad Reza Eshaghi, Tel: +989132129793, E-mail: mreshaghi@yahoo.com
Author information: Shima Shaygannia (Student), Mohammad Reza Eshaghi (Instructor), Mohammad Fazel (Instructor), Mahnaz Hashemiravan (Instructor)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Natural preservatives with high level of phenolic compounds, antioxidants and antimicrobial activities are used in mayonnaise to improve quality and safety due to their potential health benefits. Application of these compounds in production processes highlights many difficulties due to instability of their physical and chemical properties. Microencapsulation is used to address these restrictions. In this study, phenolic compounds from lemon waste were encapsulated with Persian gum (PG) and basil seed gum (BSG) as coating materials at different ratios (0:1, 1:0, and 1:1) at 15% (w/w) total biopolymer. We confirmed microencapsulation by scanning electron microscopy, and evaluate phenolic content, antioxidant activity, encapsulation efficiency, morphology, water solubility indexes, and water absorption indexes. Sample mayonnaise was prepared using microencapsulated polyphenols from lemon waste and extract (1,000 ppm of concentration), and control samples without extracts or microcapsules. All samples were subjected to chemical (measuring the peroxide, thiobarbituric acid, acidity, and color) and microbial (total count of microorganisms and Escherichia coli) analysis during 30 days of storage. BSG samples exhibited the highest antioxidant activity (61.19%) and encapsulation efficiency (70.72%), and PG/BSG microcapsules had the highest capability to prevent oxidative deterioration during storage. Addition of microcapsules led to increases in parameter b* and decreases in parameters a* and L*. In general, PG/BSG microcapsules were considered optimal samples for production of mayonnaise, since they prevented mayonnaise deterioration and exhibited antioxidant and antimicrobial properties.
Keywords: basil seed gum, encapsulation, lemon waste, mayonnaise, Persian gum
INTRODUCTION
Mayonnaise is a widely consumed food product used as seasoning for different types of food (Liu et al., 2007). Due to its structure and composition, mayonnaise is highly susceptible to chemical and microbial deterioration. Chemical deterioration includes oxidation and hydrolysis of fat and oil types, which increases the content of compounds like peroxide, leading to sourness and changes to its color, taste, and texture (Frankel et al., 2002). Mayonnaise is susceptible to deterioration due to the structure of its lipid molecules, and their interactions with other molecules in their immediate vicinity, greatly influencing susceptibility to lipid oxidation (Coupland and McClements, 1996). Mayonnaise oxidation is intensified by free radicals generated from fat oxidation with other molecules like proteins, carbohydrates, and vitamins in the emulsion. In addition, undesirable changes in pH and acidity caused by microbial or chemical activity in mayonnaise leads to microbial deterioration (Shahidi and Zhong, 2005).
Therefore, synthetic antioxidants and chemical conservatives are added to mayonnaise to control oxidation and microbial activity, which has economic advantages despite the compounds being harmful to human health (Kwon et al., 2015). For example, the food conservative sodium benzoate inhibits activity of bacteria and fungi in acidic environments (e.g., in sauces and salad seasonings). However, sodium benzoate damages cell mitochondrial DNA, consequently inactivating the cell and leading to development of the Alzheimer’s and Parkinson’s diseases (Daescu et al., 1986).
Consumption of food prepared with natural sources of phenolic compounds with a wide range of antioxidant, antimicrobial, and anti-mutagenic activities has recently increased (Kong et al., 2003). In Iran, 650,000 tons of citrus are produced annually, which accounts for 9% of the global production (Vand and Abdullah, 2012).
Lemon is a species of citrus. The large amounts of lemon waste (peels, seeds, and pulps) produced by lemon juice industries have attracted attention. Lemon exhibits antimicrobial and antioxidant activity as it contains phenolic compounds, including flavonoids, coumarin, and limonene (Altunkaya et al., 2013).
Polyphenols contain a large group of bioactive compounds, and are present in fruit, vegetables, seeds, and other parts of plants. For humans, diet is one of the most important sources of polyphenols, which exhibit antioxidant properties and roles in food preservation and human health (Shariatifar et al., 2014). These compounds are sensitive to heat, light, and oxidation due to unsaturated bands within their molecular structures (Halliwell, 2008).
The strong antioxidant activity of phenolic compounds arises from their ability to scavenge free radicals. Indeed, radical scavenging activity is attributed to the presence of hydroxyl groups that replace aromatic rings. Phenolic compounds are present in many plant-based foods that have antioxidant activity (Lakshmanashetty et al., 2010).
Destruction of phenolic compounds can be prevented by microencapsulation, a process whereby bioactive compounds are trapped by biolipids, protecting them against the environment, thus increasing stability (Belščak-Cvitanović et al., 2011). Encapsulation can be achieved by biopolymers like starch compounds, agar, alginate, chitosan, and other gums (King, 1995). Arabic gum (AB) extracted from
Persian gum (PG) is composed of galactose and arabinose and is applied as an emulsifier in acidic pH emulsion systems, high salt foods, and thermal processes. Similar to other gums, PG is water soluble and produces a sticky and viscous solution (Golkar et al., 2018).
Basil seed gum (BSG) exhibits a heterosaccharide structure that includes glucomannan, glucan, and xylan. BSG is an exclusive hydrocolloid categorized as an anionic gum and is added as an active ingredient in food formulation to increase consistency and gelation and to control microstructure, texture and aroma during storage (Dickinson, 2003; Hosseini-Parvar et al., 2010).
In the present study, we extracted phenolic compounds from lemon waste (peels and edible pulps) and microencapsulated them in gel matrixes of PG and BSG as substitutes to synthetic antioxidants and sodium benzoate in mayonnaise production. We assessed the effects of encapsulation on physical, chemical, sensory, and microbial properties of mayonnaise over one month. These phenolic compounds may represent new preservatives and flavorings for use in mayonnaise formulation.
MATERIALS AND METHODS
Materials
The primary ingredients consumed during mayonnaise preparation (oil without antioxidants, vinegar, mustard, pasteurized eggs, sugar, and salt) were purchased from a local supermarket in Isfahan, Iran. Lemons were obtained from a local farm in Shiraz. AB was provided from Sigma-Aldrich Co. (St. Louis, MO, USA). BSG and PG were purchased from a local market in Isfahan, Iran. Folin-Ciocalteau’s phenol reagent and 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) were purchased from Merck KGaA (Darmstadt, Germany) and Sigma-Aldrich Co., respectively. Methanol, hexane, potassium chloride, trichloroacetic acid, and sodium bicarbonate were purchased from Sigma-Aldrich Co.. All other chemicals or solvents used were of analytical grade.
Extraction of polyphenols from lemon waste
Polyphenols were extracted from lemon by incubating 16 g lemon with 100 mg of 70% ethanol in an ultrasonic bath (60 Hz frequency, 60°C) for 2 h. The extracts were removed by a vacuum rotary evaporator machine at 40°C. The yield extract was filtered and then incubated in a freeze dryer for 48 h (Inoue et al., 2010).
Preparation the PG and BSG
Extraction of mucilage from Basil seeds was achieved following the method described by Hosseini-Parvar et al. (2010) with slight modifications. Extraction was performed using a water/seed ratio of 50:1 at 65°C, pH 7, for 20 min whilst stirring (500 rpm, RH basic 2, IKA, Staufen, Germany). A sterile extractor equipped with a rotating rough plate was used to separate the mucilage from the swollen seeds. The mucilage was then centrifuged and filtered to remove insoluble impurities. Finally, the filtrate was freeze-dried (Dena Vacuum, Tehran, Iran) at −40°C for 24 h and stored under dry, cool, and sterile conditions.
PG was prepared according to methods described in previous studies (Fadavi et al., 2014; Golkar et al., 2015a; Golkar et al., 2015b). Briefly, impurities were removed from lighter colored PG samples and the samples were milled (AQC 109, Agromatic AG, Zürich, Switzerland) and sieved (<500 μm). PG powder was then dissolved in distilled water with stirring (23°C, 500 rpm, RH basic 2, IKA) for 2 h and stored in a refrigerator for 24 h. The solution was centrifuged at 9,000 rpm for 15 min (PIT 320R, Hettichlab, Düsseldorf, Germany) to remove insoluble residues and the supernatant was collected and dried at −40 ºC for 24 h.
Preparation of microcapsules
PG and BSG were encapsulated using polyphenol compounds as coating agents following the method described by Rutz et al. (2013) with some modifications. To prepare the microcapsules, mixtures of hydrocolloids at different PG/BSG ratios (1:0, 1:1, and 0:1) with 15% (w/w) total biopolymer were dissolved in water. Polyphenol extracts (containing 0.17 g of polyphenol) were then added so that the ratio of polyphenol extracts to wall material was 1:3. The mixture was stirred (500 rpm, RH basic 2, IKA) for 12 h at room temperature (23°C) and freeze dried (at −40°C for 24 h).
Evaluation the of microcapsule properties
Preparation of mayonnaise
Mayonnaise was produced from the oil without antioxidant (65.2%), pasteurized egg (13.85%), water (8.2%), vinegar (7.7%), salt (1.5%), and mustard (0.3%) (all amounts are stated as % w/w). First, eggs and the powdered materials (salt, sugar, mustard powder, and citric acid) were homogenized, and then oil was added until a mayonnaise emulsion had been formed. Next, vinegar was added to the mayonnaise emulsion, followed by additives (PG, BSG, or PG/BSG extracts; 1,000 ppm in all samples) and distilled water whilst mixing.
Evaluation of the mayonnaise properties
where Asample is absorbance of sample, Ablank is absorbance of reagent blank, and m is the amount of oil sample amount (mg).
Statistical analysis
Statistical analysis was performed based on completely randomized factorial design and analysis of variance (ANOVA) using SAS 9.1 software (SAS Institute Inc., Cary, NC, USA). Mean comparison tests were performed using least significant difference (LSD) methods at 5% significant level.
RESULTS AND DISCUSSION
Assessment of microcapsule properties
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Table 1 . Total phenolic content, antioxidant activity, encapsulation efficiency, WSI and WAI of lemon waste extracts and microcapsules during storage.
Samples Total phenolic content (mg gallic acid/g) Antioxidant activity (%) Encapsulation efficiency (%) WSI (g/100 g dry solid) WAI (g/100 g dry solid) BSG 16.87±0.25b 61.19±0.34b 70.72±0.76a 11.53±0.69a 24.76±0.65c PG/BSG 17.91±0.36b 58.23±0.81c 67.07±0.46b 8.33±0.27b 32.51±0.60a PG 13.92±0.33c 53.95±0.38d 65.06±0.40b 13.28±0.59a 27.11±0.03b Extract 30.50±0.25a 73.21±0.23a ? ? ? The results were expressed as mean±standard deviation of three determinations..
Different letters (a-d) in the column indicate significant differences (
P <0.05)..BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated; WSI, water solubility index; WAI, water absorption index..
Many studies have shown a direct correlation between phenolic content and antioxidant properties, whereby PG/BSG samples with the highest phenolic compound contents have lower antioxidant properties compared with BSG samples. The antioxidant properties of polyphenols arise from their redox properties, which allow them to act as reducing agents, hydrogen donators, metal chelators, and single oxygen quenchers (Piluzza and Bullitta, 2011).
The antioxidant activity of lemon extracts was higher than that of the encapsulated samples. This may be attributed to deficient extraction of the phenolic compounds in microcapsules and extract reaction with the capsule membrane, leading to reduction in antioxidant activity. da Rosa et al. (2014) obtained similar results, indicating extract and microcapsule inhibition of 93.07% and 80.38∼90.75%, respectively.
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Figure 1. SEM micrographs of encapsulated lemon waste extracts with PG (A), BSG (B), and PG/BSG (C). BSG, basil seed gum; PG, Persian gum.
In general, freeze-dried encapsulated powder had an irregular shape and flaky structure. Furthermore, micro grooves may be formed due to ice sublimation during freeze-drying. In a previous study, Pasrija et al. (2015) reported similar morphologies for freeze-dried microcapsules containing green tea polyphenols. SEM images of BSG showed that BSG has a fibril structure and contains a globular structure similar to that of the scattered cotton (Naji-Tabasi and Razavi, 2017).
Samples coated with PG/BSG showed the highest WAI (32.51±0.60%), whereas lowest WAI were observed for those coated with PG (24.76±0.65%). Therefore, samples coated with PG is less likely to be influenced by humidity and is more stable under storage conditions (Tan et al., 2015). These results suggest that the percentage of water absorption decreases with increased water solubility of the microcapsules. In a previous study, Moreira et al. (2009) obtained similar results for acerola pomace extracts. Furthermore, according to Ahmed et al. (2010) agglomeration of the encapsulated particles could contribute to a smaller WAI and, subsequently, increase WSI.
Assessing the mayonnaise properties
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Table 2 . Acid values of mayonnaise samples prepared using extracts and microcapsules of lemon waste during storage (unit: mg KOH/g).
Samples 0 10 20 30 Control 0.84±0.016abD 0.87±0.024aC 0.93±0.004aB 1.04±0.005aA BSG 0.83±0.003abB 0.81±0.006bC 0.83±0.003dB 0.87±0.003eA PG/BSG 0.85±0.004aC 0.89±0.003aB 0.90±0.005bA 0.90±0.001dA PG 0.82±0.006bD 0.90±0.006aB 0.89±0.001cC 0.91±0.003cA Extract 0.84±0.016abD 0.87±0.024aC 0.90±0.004bB 0.95±0.003bA The results were expressed as mean±standard deviation of three determinations..
Different small letters (a-e) in the same column and capital letters (A-D) in the same row indicate significant (
P <0.05) differences during the 30-day storage..Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated..
After 30 days of storage, POV peaked at 27.32±0.1 meqO2/kg oil in the control samples, with significant differences (
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Figure 2. Peroxide values (meqO2/kg) measured as peroxide and hydroperoxide concentrations in lipid extracts of mayonnaise samples prepared using extracts and microcapsules of lemon waste with PG and BSG during storage. Different letters (a-l) indicate significant (
P <0.05) differences between results. Values are mean±standard deviation of three experimental repeats. Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
Evidence of encapsulation improving bioactivity and bioavailability of polyphenols has been reported in previous studies (Fang and Bhandari, 2010). However, it has also been reported that higher concentrations of extracts may act as better pro-oxidants (Sarah et al., 2010). Rasmy et al. (2012) assessed the antioxidant effect of rosemary alcoholic extracts on mayonnaise formulation and found that applying 400 μg of extract (concentration in 1 g of mayonnaise) reduced POV compared with the control group. Moreover, similar to the increase in POVs after 4 months, mayonnaise-containing rosemary followed a slower increase compared to butylated hydroxyanisole antioxidants.
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Figure 3. Thiobarbituric acid (TBA) [mg malondialdehyde (MA)/kg] reactive substance value of lipid extracts of mayonnaise samples prepared using extracts and microcapsules of lemon waste with PG and BSG during storage. Different letters (a-j) indicate significant (
P <0.05) differences between results. Values are mean±standard deviation of three experimental repeats. Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
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Table 3 . Color measurements of mayonnaise samples prepared using extracts and microcapsules of lemon waste during storage.
Samples L* a* b* Control 91.77±0.24a ?1.31±0.03a 8.48±0.07b BSG 85.30±0.10d ?1.73±0.02c 8.51±0.13b PG/BSG 87.02±0.29c ?1.68±0.06bc 9.39±0.15ab PG 89.31±0.13b ?1.58±0.03b 10.21±0.06a Extract 91.63±0.20a ?1.51±0.01b 10.35±0.10a The results were expressed as mean±standard deviation of three determinations..
Different letters (a-d) in the column indicate significant differences (
P <0.05)..Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated..
There were no significantly differences in the L* values of the control samples and extract samples. The lowest L* values were observed for BSG sample (85.30±0.10). Furthermore, the samples containing microcapsules had lower transparency compared with the control samples. The highest a* values (red-green) were determined for the control samples (−1.31±0.03), with the lowest values recorded for the BSG samples (−1.73±0.2). In addition, the lowest b* (green-blue) value was observed for the control sample.
Adding microcapsules to prepared mayonnaise samples decreased the L* and a* indexes, and increased the b* indexes. The reduction in the L* index is due to increases in particle diameter, which reduces light differentiation and product lightness. In a previous study, McClements and Demetriades (1998) reported that mayonnaise transparency decreases as the diameters of particles diameter in the emulsion increase.
Negative a* values were reported in the mayonnaise samples. This change in color toward greenness may be due to formation of phenolic polymer compounds. Adding PG increased yellowness (b*), whereas basil mucilage induced the opposite effect due to the color pigments in the extracts and gums. Our results are in line with those of a study by Amiri Aghdaei et al. (2014) which showed decreases in transparency (L*) and yellowness (b*) by addition of
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Figure 4. Total bacterial counts of mayonnaise samples prepared using extracts and microcapsules of lemon waste with PG and BSG during storage. Different letters (a-k) indicate significant (
P <0.05) differences between results. Values are mean±standard deviation of three experimental repeats. Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated; CFU, colony-forming unit.
In all samples, the total microorganism count was <103, resulting from organic acids in the mayonnaise reducing pH and acting as an antimicrobial material (Ma and Boye, 2013). All the samples indicated negative responses to
In the current study, microencapsulation of phenolic compounds extracted from the lemon waste was performed by PG, BSG, and BSG/PG. Highest EE and antioxidant activity were observed for sample coated with BSG (70.72% and 61.19%, respectively). Furthermore, we observed increased chemical stability of microencapsulated phenolic compounds. Samples containing microcapsules reduced peroxide production, stabilized free fatty acids, and prevented increases in acidity by eliminating and preventing formation of free radicals in the mayonnaise. Mayonnaise samples prepared using PG/BSG had significantly (
ACKNOWLEDGEMENTS
The authors would like to acknowledge the Islamic Azad University (IAU), Khorasgan (Isfahan) Branch, for their valuable help, and cooperation with this project.
AUTHOR DISCLOSURE STATEMENT
The authors declare no conflict of interest.
Fig 1.

Fig 2.

Fig 3.

Fig 4.

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Table 1 . Total phenolic content, antioxidant activity, encapsulation efficiency, WSI and WAI of lemon waste extracts and microcapsules during storage
Samples Total phenolic content (mg gallic acid/g) Antioxidant activity (%) Encapsulation efficiency (%) WSI (g/100 g dry solid) WAI (g/100 g dry solid) BSG 16.87±0.25b 61.19±0.34b 70.72±0.76a 11.53±0.69a 24.76±0.65c PG/BSG 17.91±0.36b 58.23±0.81c 67.07±0.46b 8.33±0.27b 32.51±0.60a PG 13.92±0.33c 53.95±0.38d 65.06±0.40b 13.28±0.59a 27.11±0.03b Extract 30.50±0.25a 73.21±0.23a ? ? ? The results were expressed as mean±standard deviation of three determinations.
Different letters (a-d) in the column indicate significant differences (
P <0.05).BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated; WSI, water solubility index; WAI, water absorption index.
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Table 2 . Acid values of mayonnaise samples prepared using extracts and microcapsules of lemon waste during storage (unit: mg KOH/g)
Samples 0 10 20 30 Control 0.84±0.016abD 0.87±0.024aC 0.93±0.004aB 1.04±0.005aA BSG 0.83±0.003abB 0.81±0.006bC 0.83±0.003dB 0.87±0.003eA PG/BSG 0.85±0.004aC 0.89±0.003aB 0.90±0.005bA 0.90±0.001dA PG 0.82±0.006bD 0.90±0.006aB 0.89±0.001cC 0.91±0.003cA Extract 0.84±0.016abD 0.87±0.024aC 0.90±0.004bB 0.95±0.003bA The results were expressed as mean±standard deviation of three determinations.
Different small letters (a-e) in the same column and capital letters (A-D) in the same row indicate significant (
P <0.05) differences during the 30-day storage.Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
-
Table 3 . Color measurements of mayonnaise samples prepared using extracts and microcapsules of lemon waste during storage
Samples L* a* b* Control 91.77±0.24a ?1.31±0.03a 8.48±0.07b BSG 85.30±0.10d ?1.73±0.02c 8.51±0.13b PG/BSG 87.02±0.29c ?1.68±0.06bc 9.39±0.15ab PG 89.31±0.13b ?1.58±0.03b 10.21±0.06a Extract 91.63±0.20a ?1.51±0.01b 10.35±0.10a The results were expressed as mean±standard deviation of three determinations.
Different letters (a-d) in the column indicate significant differences (
P <0.05).Control, non-encapsulated samples; BSG, basil seed gum; PG, Persian gum; Extract, lemon extract free encapsulated.
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