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The avocados were purchased from a supermarket in the city of Uberaba-MG-Brazil, weighed and separated into: peel, pulp, and stone. In this material, the average weight and percentage of peel, pulp and stone were determined.
Centesimal composition of avocado pulp
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The fresh avocado pulp was characterized by physicochemical methods, according to the official methodology of the Association of Official Analytical Chemists (AOAC)[15], by determination of moisture, proteins, lipids, and ash contents. All analyzes were performed in triplicate.
Water content determination
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Ten gram of the sample was weighed in a porcelain capsule and the sample was heated for 6 h at 105 °C. The sample was then cooled in a desiccator to room temperature, heated and cooled repeatedly until a constant mass was obtained, and the moisture percentage was calculated according to Eqn (1).
$ Umidity\;{\text%}=\dfrac{\mathrm{S}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}\;\mathrm{m}\mathrm{a}\mathrm{s}\mathrm{s}\left(\mathrm{g}\right)-Dry\;Sample\;mass\left(g\right)}{\mathrm{s}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}\;\mathrm{m}\mathrm{a}\mathrm{s}\mathrm{s}\;\left(\mathrm{g}\right)}\times 100 $ (1) Protein determination
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One gram of the sample was weighed, transferred to a Kjeldahl flask, 25 mL of concentrated sulfuric acid (96%) and ~6 g of the catalytic mixture was added. The sample mixture was heated on an electric plate until the solution turned blue-green and was free of undigested material. The sample was subsequently heated for 1 h and allowed to cool. The material from the Kjeldahl flask was quantitatively transferred to a distillation flask, 10 drops of phenolphthalein indicator and 1 g of zinc powder were added. The flask was immediately coupled to the distillation equipment and connected to an erlenmeyer flask containing 25 ml of 0.05 M sulfuric acid and 3 drops of the methyl red indicator. Sodium hydroxide solution (30%) was added to the vial containing the digested sample, which was then heated to boiling and distilled to obtain ~250 ml of distillate. The excess sulfuric acid was titrated with sodium hydroxide (0.1 M) using methyl red indicator. The percentage of proteins was calculated according to Eqn (2).
${ Proteín\;{\text%}=\dfrac{\mathrm{V}\mathrm{o}\mathrm{l}\mathrm{u}\mathrm{m}\mathrm{e}\;\mathrm{s}\mathrm{p}\mathrm{e}\mathrm{n}\mathrm{t}\;\mathrm{o}\mathrm{n}\;\mathrm{t}\mathrm{h}\mathrm{e}\;\mathrm{t}\mathrm{i}\mathrm{t}\mathrm{r}\mathrm{a}\mathrm{t}\mathrm{i}\mathrm{o}\mathrm{n}\left(\mathrm{m}\mathrm{L}\right)\times 0.14 \times Correction\;factor*}{\mathrm{S}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}\;\mathrm{w}\mathrm{e}\mathrm{i}\mathrm{g}\mathrm{h}\mathrm{t}} }$ (2) * Correction factor considered equal to 6.25.
Lipid determination
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Approximately 2 g of the dry sample was weighed and transferred to the Soxhlet-type extractor device and kept under heating for 6 h (2−3 drops per second). The material was removed from the extractor and the flask with the extracted residue was transferred to an oven at 105 °C, for ~3 h. The sample was cooled in desiccator to room temperature. The percentage of lipids was calculated according to Eqn (3).
$ Lipid\;{\text%}=\dfrac{\mathrm{L}\mathrm{i}\mathrm{p}\mathrm{i}\mathrm{d}\;\mathrm{m}\mathrm{a}\mathrm{s}\mathrm{s}\;\left(\mathrm{g}\right)}{\mathrm{S}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}\;\mathrm{m}\mathrm{a}\mathrm{s}\mathrm{s}\;\left(\mathrm{g}\right)}\times 100 $ (3) Ash determination
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Approximately 5 g of the sample was weighed in a porcelain capsule and elevated to a muffle at 550 °C. After cooling, the sample was weighed, and the percentage of ash was calculated according to Eqn (4).
$ Ash\;{\text%}=\dfrac{\mathrm{A}\mathrm{s}\mathrm{h}\;\mathrm{w}\mathrm{e}\mathrm{i}\mathrm{g}\mathrm{h}\mathrm{t}\;\left(\mathrm{g}\right)}{\mathrm{A}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}\;\mathrm{w}\mathrm{e}\mathrm{i}\mathrm{g}\mathrm{h}\mathrm{t}\left(\mathrm{g}\right)} \times 100 $ (4) Extraction of avocado pulp
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The fresh avocado pulp was submitted to drying in an oven at 60 °C and was ground to obtain a bran. Then, the avocado pulp bran was extracted by solvent extraction with pure ethanol over 2 d. After filtration, the extract was concentrated in a rotary evaporator.
Gas chromatography with mass spectrometry (GC-MS)
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The GC-MS analysis was performed as described by Posetti et al.[16]. The apparatus employed was a Shimadzu 2010 High Resolution Gas Chromatograph coupled to a Mass Spectrometry Detector. An Agilent DB-5MS column (30 m × 0.25 mm − 0.25 μm) was employed. The following run conditions were used: Injector Temperature: 220 °C, Splitless Injection Mode, 2 min sampling time, linear velocity 15.7 Psi Pressure, Total flow of 19.4 mL·min−1, Column flow of 1.49 mL·min−1, Linear velocity of 45.0 cm·s-1, Purge flow of 3.0 mL·min−1, Split ratio of 10, Gradient Mode column temperature of 80−280 °C. The Mass Spectrometry Detector parameters were 200 °C ion source temperature, 280 °C interface temperature, 3 min solvent cut-off time, 3 min initial detection time, a detection time of 17.0 min, SCAN acquisition mode, 0.25 s acquisition time, SCAN mass/charge ratio (m/z) from 40 to 600 and 1 μL injection volume[16].
Minimum inhibitory concentration (MIC)
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MIC determination was performed using 96-well microdilution plates (12 columns and 8 rows) and three microorganisms, Bacillus cereus (ATCC11778), Staphylococcus aureus (ATCC 29213) and Escherichia coli (ATCC 35218), according to Methods for Dilution Antimicrobial Susceptibility Tests[17]. Initially, the dilutions were carried out in test tubes where BHI broth and the tested extract were added. The dilutions were made in the following percentages: 100% extract, 95%, 90%, 85%, thus up to 5% extract. In each well of the plate, 300 µL of the previously diluted solution was added to the test tubes except for the last two columns, where the wells were only filled with BHI broth, for the positive control (using chlorhexidine 2%) and the control negative. After all wells were filled, the microorganisms were added, each in its respective plate, only the last column did not receive the microorganism, as this was the negative control. The plates were incubated at 37 °C for 24 h. The MIC is the lowest concentration that completely inhibits the growth of the microorganism, that is, in which turbidity is not observed in the medium in the well[4, 16, 17].
Preparation of coatings
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In the development of coatings, avocado pulp extract at 5% was used. The chitosan-based edible coating was elaborated according to de Araújo et al.[18]: 2% (m/v) Chitosan were dissolved in 1% (v/v) acetic acid and stirred under heating (40 °C) for 6 h. Subsequently, 0.75 mL of glycerol/q of chitosan was added, and the solution was stirred for another 30 min. After this period, 5% (v/v) avocado pulp extract was added.
The alginate-based edible coating was prepared according to Santos et al.[4] and de Araújo et al.[18]: 1.29% (w/v) of sodium alginate was dissolved in distilled water under heating (70 °C) and mechanical stirring until the mixture was clear. Then, 1.16% (m/v) of glycerol and 5% of the avocado pulp extract were added. To promote the crosslinking of the polymer matrix, a solution containing 2% (m/v) of calcium chloride, 1% (m/v) of ascorbic acid and 1% (m/v) of citric acid was used.
Application of the edible coatings
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The edible coatings were applied to 'Fuji' variety apples, purchased in a supermarket in the city of Uberaba, Brazil. The chosen apples were ripe and selected manually, analyzing the redness, average size and if there were no signs of injury. The fruits were washed and sanitized in 0.5% chlorinated water (v/v) for 15 min, then peeled and cut into slices of approximately 25 g. The apple slices were submerged in the coating solutions and left to rest to allow for dripping. The fruits coated with the alginate-based coating were submerged in the solution 2% (m/v) of calcium chloride, 1% (m/v) of ascorbic acid and 1% (m/v) of citric acid to promote polymeric crosslinking. Once dried, samples were packaged separately in non-toxic plastic bags, sealed and stored at 3 °C for the analysis period (15 days performing microbiological analysis every 5 d). Four treatments were obtained:
• AA1 – apple + alginate-based coating without the extract.
• AA2 – apple + alginate-based coating with the extract.
• AC1 – apple + chitosan-based coating without the extract.
• AC2 – apple + chitosan-based coating with the extract.
Microbiological evaluation of the samples (shelf life)
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For the standard plate count, 25 g of the sample were added to 225 mL saline solution (0.1%), followed by homogenizing and proceeding with the serial dilution up to 10-4. Afterwards, 1 mL of each dilution was inoculated into sterile, empty Petri dishes, pouring 20 mL of molten Plate Count Agar medium. The medium was homogenized with the inoculum through movements in the shape of a figure eight and, after the medium had solidified, the plates were incubated at 37 °C for 24 h[19]. For the determination of thermotolerant coliforms, the technique of multiple tubes by the most probable number (MPN/g) was used. A 10-1 dilution was performed, homogenizing 25g of sample with 225 mL of peptone water (0.1%) and from the serial dilutions (10-2 and 10-3), 1 mL of each dilution was seeded. The medium used was 0.1% peptone water and incubated at 35 °C for 24 h. From the reading of the positive tubes (with growth and gas production in the Durham tubes), confirmation was performed with Brilliant Green Bile Broth under the same time and temperature conditions recommended by the American Public Health Association (APHA)[20]. The determination of the Most Probable Number of Thermotolerant Coliforms was carried out using the technique of multiple tubes, with Broth Escherichia coli (EC). For positive tubes for thermotolerant coliforms, culture subcultures were performed into tubes with EC Broth. After sowing, the sample was incubated at 37 °C for 24 h, in a water bath, with a series of three tubes for each subculture[21]. Subsequently, an elevation of the positive EC Broth tubes was striated on plates containing the Eosin Methyl Blue medium and incubated at 37 °C for 24 h to confirm the growth of Escherichia coli characterized by the growth of black flowering colonies. As for the evaluation of Salmonella ssp, 25 g of each sample were homogenized in 225 mL of broth for Salmonella enrichment and incubated at 30 °C, after 20−24 h 1 mL aliquots were transferred to tubes containing 9 mL of Tetrathionate Broth Base and incubated at 30 °C for 20−24 h. The sample was then, striated to a Petri dish containing XLD Agar and incubated at 37 °C for 24 h[19, 22].
Sensory analysis
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The sensory analysis was approved by the Research Ethics Committee of UFTM (process number 2,108.891), using the acceptance test employing a 9-point hedonic scale that contains the defined terms situated between 'I really liked' and 'I disliked extremely', as described by Santos et al.[4] and Posetti et al.[16]. The sensory analysis was performed in a laboratory that has a specific area for sample preparation and individual, climate-controlled booths. The tasters were instructed to read the Informed Consent Form and after clarifying any doubts that they had about the object of the study, the form was signed in two copies, one of which remained with the taster and the other with the researcher. This analysis involved the participation of 50 untrained tasters aged between 18−63 years, composed of university employees and students. The samples were characterized microbiologically and physiochemically before sensory analysis. The apple samples slices of approximately 25 g (a coating-less control sample apple without any coating), an AA1 sample, an AA2 sample, an AC1sample and an AC2 sample) were served individually in plastic dishes, and duly identified with numerical codes of three random digits. Samples were served simultaneously and were at 8 °C. Tasters were instructed to identify the sample codes on the sensory analysis form, taste the samples from left to right, arranged on the tray, record the score according to the hedonic scale score for each attribute: color, aroma, texture and taste and finally rinse your mouth with water before tasting the next sample.
Statistical analysis
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Data were analyzed using Analysis of Variance (F test) at 5% probability and once the significant effect of the treatment was verified, the Tukey test was applied at 5% probability. Statistical analyzes were performed using the SISVAR computer program, developed by Ferreira[23].
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The raw material characterization is important to observe how the fruit varies, besides being possible to see how significant the by-products are. The average percentage of each component and the average values and standard deviation of pulp, peel, seed and fruit, where the accuracy of the measuring equipment was 0.01g (Table 1). Emphasizing that the variety of avocado used was Fortuna.
Table 1. Average weight and average percentage of components.
Attributes Seed mass
(g)Pulp mass
(g)Pell mass
(g)Total mass
(g)Average weight 105.05 ±
6.07452.56 ±
39.0051.34 ±
0.05609.06 ±
45.19Average percentage 17.25% 74.30% 8.45% − Proximate composition of the avocado pulp
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The samples were separated and the analyzes were performed in triplicate, the precision of the measuring equipment was 0.0001 g (Table 2).
Table 2. Results of the proximate composition of the Fortuna avocado pulp.
Sample Water content (%) Protein (%) Dry matter lipids (%) Whole matter lipids (%) Ashes (%) Pulp 83.4 ± 0.19 1.16 ± 0.12 53.62 ± 1.67 8.89 ± 0.26 0.53 ± 0.011 Avocado pulp extraction, Minimum Inhibitory Concentration (MIC) and evaluation of the chemical composition of the obtained extract
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The extraction had a yield of approximately 8%, considering the input of fresh raw material, that is, with high water content. The minimum inhibitory concentration values for the extract obtained against Escherichia coli, Staphylococcus aureus and Bacillus cereus are shown in Table 3.
Table 3. MIC of the studied extract.
Avocado pulp extract Escherichia coli Bactericide 80% Bacteriostatic 75% Staphylococcus aureus Bactericide 80% Bacteriostatic 75% Bacillus cereus Bactericide 45% Bacteriostatic 40% Regarding the chemical composition, 12 peaks were obtained in the GC analysis that were identified by MS and Table 4 shows the compounds present in the avocado pulp extract.
Table 4. Chemical composition of avocado pulp extract.
Retention time (min) Compound Structure 7.825 (6E)-3,7,11-trimethyl-1,6,10-dodecatrien-3ol 10.154 2-octylfuran 11.758 palmitic acid 13.433 ci-vaccenic acid 13.933 1,2-decanediol 15.025 3-methyl-2-(2-methylene-cyclohexyl)-butan-2-ol 15.504 (13,14-epoxy)-tetradec-11-en-1-ol acetate 15.651 7-octene-1,2-diol 15.942 2,3,4-trimethyl-5-hexen-3-ol 16.225 2,3,6-trimethyl—7-octen-3-ol 16.425 10-methyl-(11E)-tridece-1-ol acetate 16.762 O-methyloxime-(2E)-nonadecanone Microbiological evaluation – shelf life
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The samples of fresh apples (without any coating, only minimally processed) and coated with alginate or chitosan-based coating with and without avocado pulp extract, were evaluated for 15 d, every 5 d and the results are shown in Table 5.
Table 5. Standard count on plates.
Day 0
(CFU)Day 05
(CFU)Day 10
(CFU)Day 15
(CFU)In natura apple 3.2 × 103 5.7 × 105 1.5 × 106 2.0 × 106 AA1 < 3.0 < 3.0 2.0 × 104 1.0 × 105 AA2 < 3.0 < 3.0 1.0 × 102 3.8 × 102 AC1 8.0 × 102 5.0 × 104 3.5 × 105 6.8 × 105 AC2 < 3.0 3.0 × 104 4.8 × 105 6.6 × 105 Sensory analysis
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The statistical analysis of the results obtained in the sensory analysis are summarized in Table 6. It was observed that the coated apples were well accepted by the panelists, except for sample AC1 (apple + chitosan-based coating without extract) which obtained in the global evaluation a mean less than 5, which symbolizes neither liked nor disliked in the hedonic scale used.
Table 6. Averages, standard deviations and results of statistical tests to compare the evaluated attributes.
Samples Attributes evaluated Apparency Aroma Texture Flavor General AA1 8.00 ± 1.28A 6.88 ± 1.55A 7.74 ± 1.47A 6.80 ± 1.88A 7.26 ± 1.23A AA2 7.86 ± 1.78A 6.62 ± 1.59A 7.44 ± 1.45A 6.60 ± 2.12A 7.04 ± 1.52A AC1 4.68 ± 1.94Ba 4.94 ± 1.72B 5.88 ± 2.09B 4.88 ± 2.09B 5.14 ± 1.59B AC2 4.42 ± 1.75Ba 4.76 ± 2.07B 5.76 ± 2.40B 4.24 ± 2.33B 4.76 ± 1.80B Control (in natura apple) 5.94 ± 1.89Bb 6.36 ± 1.69A 6.94 ± 1.70A 6.90 ± 1.64A 6.54 ± 1.33A Legend: Means followed by the same letter between lines do not differ significantly from each other, at 5% probability, by Tukey's test. -
The edible coatings prepared from alginate showed the best performance in the various criteria evaluated. Regarding shelf life, samples with alginate-based coating had the best results, and the sample coated with alginate-based coating and incorporating P. americana (avocado) pulp extract showed the best result, 15 d of shelf life. Sensorially, as the alginate-based coatings were able to inhibit enzymatic browning, considerably improving the appearance of the minimally processed apple and obtained the best results in all aspects evaluated in the sensory analysis. It can be said then, the alginate-based edible coating incorporating P. americana pulp extract is a viable solution for the preservation of minimally processed apples.
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About this article
Cite this article
Ribeiro AAP, Sanfelice RC, Malpass GRP, Okura MH, Malpass ACG. 2023. Shelf life and sensory analysis comparison of alginate and chitosan edible coating incorporating avocado extract applied to minimally processed apples. Food Materials Research 3:26 doi: 10.48130/FMR-2023-0026
Shelf life and sensory analysis comparison of alginate and chitosan edible coating incorporating avocado extract applied to minimally processed apples
- Received: 19 April 2023
- Accepted: 23 July 2023
- Published online: 26 December 2023
Abstract: The purpose of the present study was to evaluate Persea americana (avocado) in relation to the pulp composition, moisture content, lipids present, proteins, and ash. The P. americana pulp extract was prepared by solvent extraction using pure ethanol, and the chemical composition of the extract was evaluated by GC-MS. The resulting extract was subsequently incorporated in alginate and chitosan-based edible coatings on minimally processed Fuji apples. The edible coatings were evaluated microscopically, for shelf-life improvement and by sensory analysis. The alginate-based coatings were able to inhibit enzymatic browning, improve the appearance of the minimally processed apple samples and obtained the best results in all aspects evaluated in the sensory analysis. The incorporation of P. americana pulp extract contributed to the improvement the shelf life of minimally processed apples for 15 days and the fruits covered with the alginate-based edible coating incorporated with P. americana pulp extract had the best scores in acceptance and appearance.
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Key words:
- Edible coatings /
- Minimally processed apples /
- Persea americana /
- Shelf life