MINISTRY OF EDUCATION AND TRAININGNONG LAM UNIVERSITY - HO CHI MINH CITY Faculty of Chemical Engineering and Food Technology THE DEVELOPMENT OF PLANT-BASED MEATBALL FROM SOLID WASTE OF S
Extraction of total phenolic content from split gill mushroom
Cutting Extract at 120°C for 10 minutes
Drying for 3 hours at li
Figure 3.2 Autoclave extraction of total phenolic content from split gill mushroom
The extraction of total phenolic content from split gill mushrooms was conducted using an autoclave method Initially, the mushrooms were washed, chopped into small pieces, and dried at 60 °C for three hours The extraction involved mixing the dried mushrooms with distilled water in a 1:6 ratio, followed by incubation at 120 °C for ten minutes in the autoclave After cooling to room temperature for thirty minutes, the mixture was filtered to separate the liquid and solid components.
The solid fraction of the split gill mushroom was dried at 60°C for four hours in a dehydrator, then ground into a fine powder using a grinder The resulting split gill mushroom powder was subsequently packaged in plastic bags and stored in a desiccator until ready for use.
Solid fraction of SGM extraction
Figure 3.3 Preparation of split gill mushroom powder
The study examined and compared various properties of both liquid and dried solid fractions, focusing on key measurements such as color, pH, and particle size Additionally, a proximate analysis was conducted to assess moisture, fat, protein, fiber, and ash content.
Application of split gill mushroom powder in the formulation of plant-based
This experiment aimed to evaluate how varying ratios of split gill mushroom (SGM) powder affect the quality of plant-based meatballs Conducted as a one-factorial randomized study, it included five treatments: a control group with 0% SGM, and groups with 5%, 10%, 15%, and 20% SGM added The specific formulations for the meatball samples are detailed in Table 3.1.
Table 3.1 Experimental design for the development of a plant-based meatballs formula
Raw material Control 5% SGM 10%SGM 15%SGM 20% SGM
Mung bean Vua 7.3 7.3 7.3 hd Chickpea(g) 5.5 55 5.5 5.5 5.5
Water (g) 30 45 60 72 90 Ice (g) 35.6 35.6 35.6 35.6 35.6 Pepper (g) 0.1 0.1 0.1 0.1 0.1 Sea salt (g) 0.5 0.5 0.5 0.5 0.5 Sugar (g) 1:5 15 1.5 1.5 15
The preparation of plant-based meatballs involved several key steps First, soy protein isolate was soaked in water for 30 minutes Then, chickpea flour was dissolved in water and heated for 6 minutes to minimize its yellow color and odor Afterward, all ingredients were combined in a bowl and mixed for 5 minutes in a mixing machine During this process, konjac flour, water (with mushroom powder), and ice were gradually incorporated Finally, the mixture was shaped into small balls to complete the meatball preparation.
The meatballs, measuring 2.5 cm in diameter and weighing 11.4 grams, were boiled in water at 100°C for four minutes After boiling, they were cooled in cold water for 50 seconds and allowed to drain at room temperature for one and a half hours.
Soy isolate protein Sulit ill mnmiissia Chickpea (dissolved in
(soaked in water for 30 anil water and boiled at minutes) 60°C for 6 minutes)
⁄ ` Konjac flour and ice (slowly added while mixing in the mixing machine)
Dough was formed into small balls
Small balls (boiled at a temperature of 100°C for 4 minutes) y,
In the preparation of plant-based meatballs, all samples underwent analysis for color (L*, a*, and b* values) and various texture properties, including hardness, adhesiveness, springiness, cohesiveness, gumminess, and chewiness The highest quality samples were then chosen for sensory evaluation, which significantly influenced the final selection for further analysis and potential product development Ultimately, the two top-quality samples were selected for proximate analysis, assessing moisture, fat, protein, fiber, ash, carbohydrate content, and total phenolic content.
Methods for anal ysiSicansccsepscanmam rN RRES 29
Sensory quality/ Preference test of plant-based meatball
A total of 30 untrained panelists were recruited from Kasetsart University in Bangkok, Thailand, for the study The selection criteria included individuals aged between 18 and 60 years who regularly consumed minced meatballs and had no history of food allergies Panelists with asthma or any allergies were excluded from participation.
Each treatment consisted of one whole meatball placed in a labeled plastic bag, with samples served in cups coded with random 3-digit numbers to minimize bias Panelists were given drinking water to cleanse their palates between tastings They were instructed to visually assess the product's appearance and color before tasting, followed by scoring the samples for odor, taste, firmness, and overall liking on a 9-point hedonic scale, as outlined by Meilgaard et al (1999).
Chemical properties of split gill mushroom material after extraction
The moisture content was estimated using AOAC protocol (AOAC,
Moisture content was determined by calculating the weight difference before and after drying samples in an oven at 105 °C for 5 hours and 30 minutes This heating and cooling cycle was repeated until a stable weight was reached, with the weight loss after drying representing the moisture content.
Moisture (%) = (initial weight — final weight) x 100/initial weight of sample
Crude fat content was measured using the Soxhlet extraction method, where petroleum ether (boiling point 40 - 60 °C) was added to each sample in the extraction apparatus The extraction process lasted for 3 hours using a Biichi Labortechnik AG Extraction Unit E-812/E-816 HE, after which the ether was evaporated to dryness The fat amount was determined by calculating the weight difference of the flask before and after the ether was removed (AOAC, 2012).
W, = weight of flask with fat (g).
Crude protein content was measured by the Kjeldahl method One gram of ground sample from each of the mushroom types was digested in the Labtec
The TM Line Digestion Systems incorporate a scrubber unit that utilizes 98% sulfuric acid, followed by steam distillation After digestion, the sample is processed in the KjeltecTM8100 distillation unit The resulting solution is titrated to a pink or wine-red hue with 0.1 N hydrochloric acid, allowing for the calculation of the protein percentage using a specified formula.
V, = volumeof standard HCI acid used to titrate a sample (Nollet);
V, = volume of standard HC] acid used to titrate a blank (Nollet);
M = molarity (0.1) of standard HCI acid (N);
W = weight (g) of dried sample used.
A sample weighing one gram was placed in crucibles, followed by the addition of 1 gram of Celite 545, and analyzed using the Fibertec TM 8000 Auto Fibre Analysis System Initially, a moisture-free and ether-extracted crude fiber sample, composed of cellulose, was digested with dilute sulfuric acid (H₂SO₄) and subsequently with dilute potassium hydroxide (KOH) solution The undigested residue obtained post-digestion was ignited, and the weight loss recorded after ignition was identified as crude fiber Finally, the crucibles were positioned in a muffle furnace for further analysis.
500 °C for 5 h, cooled to room temperature, and then reweighed to find the fiber content percentage (AOAC, 2012).
W, = weight of crucible with fiber (g);
W, = weight of crucible with ash (g).
The ash content of dried split gill mushroom powder was determined following the AOAC protocol (AOAC, 2012) A 1 g sample of the powdered mushroom was placed in a crucible and subjected to combustion in a muffle furnace at 555 °C for 6 hours After cooling, the crucibles were stored in desiccators and weighed to obtain the final results.
W, = weight of crucible with ash (g).
The content of the available carbohydrate was calculated by difference according to the following equation:
Carbohydrate content (%) = 100 — [(ash + moiture + protein + fat) %] 3.4.5.7 Energy value
The energy values of the samples were calculated using following formula (AOAC, 1990):
Energy (Kceal/100 g) = (Crude protein x 4) + (Total carbohydrate x 4) + (Crude fat x 9)
Folin 10%: Take 10 ml of folin into a volumetric flask, then add water to 100ml The 10% folin solution is stored in the refrigerator and in light- blocking conditions.
Methanol 80%: Weigh 625 ml of 99.7% pure methanol, add 49.375 ml of distilled water to a 250 ml volumetric flask.
Na2CO3 7.5%: Weigh 7.5 g Na2CO3, makeup to 100 ml into the corresponding volumetric flask with distilled water.
A sample of two grams was dissolved in 20 ml of 80% methanol and vortexed for two minutes to achieve homogeneity (Melia et al., 2021) The solution was then ultrasonicated at 30 °C for 15 minutes and centrifuged at 4000 rpm for 10 minutes to collect the first supernatant For the second extraction, 15 ml of methanol was added to the pellet, and the resulting mixture was centrifuged again for 10 minutes to obtain the final supernatant for analysis.
The total phenolic content (TPC) was measured by mixing 400 µL of a filtered sample with 2 mL of 10% Folin reagent and allowing it to stand for 5 minutes in darkness Following this, 1.6 mL of Na2CO3 was added to the mixture, which was then incubated for 30 minutes at room temperature in the dark Absorbance readings were taken at 765 nm, with triplicate measurements performed for each sample to ensure accuracy.
The formula used to determine TPC
TPC: Total polyphenol content (mg GAE/g DM);
P: Phenolic content from the standard curve equation (ug GAE/ml);
V: volume of sample solution (Nollet);
D: dilution factor; m: weight of sample (g);
This study utilized a completely randomized design (CRD) to evaluate the effects of varying concentrations of SGM in meatballs The treatments included five different levels of SGM substitution: 1) no SGM, 2) 5% SGM (5 g SGM powder + 15 g water), 3) 10% SGM (10 g SGM powder + 30 g water), 4) 15% SGM (15 g SGM powder + 45 g water), and 5) 20% SGM (20 g SGM powder + 60 g water).
Data analysis was conducted using Microsoft Office Excel 2021 and Minitab 17, employing one-way analysis of variance (ANOVA) to assess the effects of treatments The differences among treatment means were evaluated with the Tukey test, as outlined by Steel et al (1997), with significance set at P < 0.05.
RESULTS AND DISCUSSION cccccsscsssssssssssssssssesessssssssesssees 36 4.1 Properties of liquid and solid fractions after extraction of total phenolic content IrGiBi,SBIIEJDIIlRUESHTOGHuusreeeenebiniastiitoi EitdSdEEGSIIHGENSSHGSSSNGERONSISEEEIGGRSSEIGSUISBUEHS.SISOG.HISSI4E018 36 4.1.1 Color, pH, and dwg (geometric mean diameter) measurement 3Ó 4.1.2 Proximate analysis of liquid and solid fractIons
Yield of split gill mushroom powder after drying
The production yield of dried split gill mushroom powder (Schizophyllum commune) was determined to be 29.02%, as shown in Table 4.3 This yield falls within the typical range of 20-30% for dried samples For comparison, the drying yield of Phyllanthus amarus has been reported to range from 15.32% to 23.84% (Nguyen et al., 2015).
Table 4.3 Yield of dried split gill mushroom
Parameter Value Drying temperature 60°C Time 4 hours Dried Yield 24.89 %
The drying yield of split gill mushrooms (Schizophyllum commune) is affected by initial moisture levels, drying methods, environmental conditions, and the mushrooms' specific traits Consequently, the yield can vary significantly based on these factors Additionally, drying may lower the total bitterness associated with free amino acids, indicating an enhancement in the taste characteristics of the mushrooms post-drying.
In 2023, drying significantly enhanced the organic acid content in S commune, with succinic acid being the predominant organic acid found in the mushroom Utilizing an appropriate processing method can yield S commune with an improved umami flavor and extended shelf life, making it a valuable nutritive ingredient for food production.
4.2 Application of split gill mushroom powder in the formulation of plant- based meatball
4.2.1 Color measurement of plant-based meatball with different split gill mushroom powder ratios
The color of plant-based meatball with different split gill mushroom powder ratios was measured and presented in Table 4.4 and Figure 4.2.
Table 4.4 Color parameters of the plant-based minced meatball with different levels of SGM powder
Mean + SD with different superscripts in each column are significantly (p < 0.05) different
Figure 4.2 Color of plant-based meatball with different split gill mushroom residues
The addition of split gill mushroom (SGM) powder to plant-based meatballs resulted in a lower L* value compared to the control, likely due to the high dietary fiber content in mushrooms, which reduces light reflection and causes surface drying As the SGM powder ratio increased, the a* value, indicating redness, gradually rose, while the b* value showed no significant trend These changes suggest that higher SGM powder levels contribute to a darker appearance and enhanced redness, potentially affecting the product's visual appeal and consumer acceptance, influenced by the concentration of SGM powder and its effects on the product matrix.
4.2.2 Texture properties of plant-based meatball with different split gill mushroom powder ratios
The texture profile analysis results for plant-based meatballs reveal that texture significantly impacts the overall eating experience Achieving an ideal texture involves careful selection of ingredients, processing methods, and formulation techniques Data indicates that the level of SGM powder added influences all textural parameters, with mushroom-based meatballs demonstrating lower texture scores compared to the control This suggests that incorporating mushrooms into plant-based meatballs may lead to a reduction in textural quality.
In 2023, research by Mena et al highlighted the importance of meat texture, emphasizing that hardness significantly influences consumer preferences Older consumers tend to prefer meat products with lower hardness levels, indicating a shift in texture preference among different age groups.
The hardness of the meatball was measured at 8.14 to 17.66 Newtons, aligning with Hirunyophat's (2023) findings, which reported hardness values of 15.63 and 15.89 Newtons for plant-based meatballs containing 5% and 10% pea protein isolate, respectively.
Table 4.5 Texture parameters of the plant-based meatball with different
Measurements Control 5% SGM 10% SGM 15% SGM = 20% SGM Hardness (N) 17.66+ 2.60? 15.794+2.15* 12.614 1.62° 10.75+2.37° 8.14+1.27 nia thuyng -0.09 + 0.04 -0.4140.48 -062+0.44° -1.22+ 0.56° -2.40 + 0.554
Mean + SD with different superscripts in each row are significantly (p < 0.05) different
4.2.3 Sensory test of plant-based meatball with different split gill mushroom powder ratio
Figure 4.3 The spider web of sensory liking of the plant-based meatball with different SGM powder ratios
In a sensory evaluation of plant-based meatballs using a 9-point hedonic scale, the content of mushroom residue emerged as the most influential factor affecting color, appearance, and firmness The results of the sensory analysis, which examined various ratios of SGM powder added to the meatballs, are illustrated in Figure 4.3.
The study revealed that the plant-based meatball control achieved the highest color score of 6.867 ± 1.907, while the 5% SGM and 10% SGM treatments scored 6.400 ± 1.476 and 6.100 ± 2.264, respectively There were no significant differences (p > 0.05) in color between the 5% and 10% SGM treatments, but the 15% SGM treatment exhibited the lowest color score at 5.167 ± 2.379 Color significantly affects consumer preference for meatball products (Gedikoglu and Clarke, 2019) Additionally, the 15% SGM treatment had the lowest scores for appearance and firmness, with values of 4.667 ± 2.279 and 2.933 ± 1.596, respectively The darker color and softer texture of the split gill mushroom particles contributed to these lower scores in color, appearance, and firmness.
45 meatball, while there were no significant differences (p > 0.05) in the odor, chewiness, taste, and overliking of the meatball product.
The odor of mushrooms does not impact the quality of the final product For further analysis, the control and 10% SGM treatments were chosen due to their superior liking scores and potential for enhanced nutritional value.
4.2.4 Chemical properties of plant-based meatball with two different split gill mushroom powder ratios (0% SGM and10% SGM)
4.2.4.1 Proximate analysis of plant-based meatball with 0% SGM and 10% SGM
The proximate results of plant-based meatballs with 10% SGM and without SGM are shown in Table 4.6.
Table 4.6 Proximate composition of plant-based meatball product with
Measurements 0% SGM meatball 10% SGM Meatball Moisture (Yowb) 74.91 +0.12> 77.30 + 0.058
Mean + SD with different superscripts in each column are significantly (p < 0.05) different
The addition of split gill mushroom (SGM) powder significantly increased the moisture content of the product due to its high water-holding capacity, measured at 396 ± 14.26 g/g (Acharya et al., 2016) This moisture level, at 9.831 ± 0.33%, remains within safe limits to inhibit microbial growth Additionally, the protein content of the product was found to be 14.98 ± 0.48%, surpassing that of commercially available plant-based meatballs, which had a protein content of 8.77 ± 0.02 g (Hirunyophat, 2023).
The addition of 10% SGM resulted in a slight decrease in fat (5.18 + 0.33% db) and energy content (392.50 + 1.56 kcal/100 g) compared to the control (0% SGM), which had higher values of 7.33 + 0.98 % db fat and 404.99 + 6.70 kcal/100 g energy The low ash percentage of SGM powder contributed to a reduction in the overall ash content of the product While the inclusion of 10% SGM increased fiber content, it did not create a significant difference when compared to the control The control meatball exhibited higher protein levels due to the use of soy protein isolate, leading to a dilution effect in the SGM product that slightly decreased its protein content; however, no significant differences in protein levels were observed overall Ultimately, the new product development maintains comparable energy levels to the control while offering a higher carbohydrate content.
4.2.4.2 Total phenolic content (TPC) of plant-based meatball with 0% SGM and 10% SGM
The total phenolic content of the control meatball (0% SGM), and the developed product (10% SGM) are presented in Figure 4.4.
Figure 4.4 Total phenolic contents of developed plant-based meatball product from split gill mushroom
Different letters indicate significant differences (p < 0.05)
SGM meatballs exhibited a significantly higher total phenolic content of 1.01 ± 0.10 mg GAE/g of dry extract, compared to the control meatball's 0.39 ± 0.04 mg GAE/g, highlighting the impact of SGM powder on enhancing phenolic content This increase is crucial as the total phenolic content is linked to antioxidant properties, supported by studies such as Panusa et al (2013), which found a strong correlation between phenolic content and antioxidant activity in spent coffee grounds Additionally, Bochnak-Niedzwiecka et al (2022) demonstrated that the phenolic content in protein-rich powdered beverages was significantly enhanced by the addition of lentil proteins and flaxseed gum, further influencing their antioxidant capabilities.
The plant-based meatball incorporating 10% SGM exhibited a notably higher total phenolic content than the control formula lacking SGM, while maintaining similar proximate values These findings suggest that the solid fraction of SGM obtained through autoclave extraction is a valuable ingredient for creating plant-based meatballs.
CONCLUSIONS AND RECOMMENDATIONS
The extraction of total phenolic compounds from split gill mushrooms revealed that the liquid fraction contained the highest phenolic content, measuring 22.41 ± 1.23 mg GAE/g of dry extract, compared to just 3.45 ± 0.19 mg GAE/g in the solid fraction This research suggests that the solid waste from autoclave extraction of split gill mushrooms can be effectively utilized as an ingredient in the production of plant-based meatballs.
The ideal formulation for plant-based meatballs consists of 10% SGM, incorporating ingredients such as 7% split gill mushroom dried powder, 5% soy protein isolate, 5% mung bean, 4% chickpea, and various other components including tapioca flour, konjac flour, and baking powder This optimized recipe also includes water, ice, pepper, sea salt, sugar, and sunflower oil Notably, the plant-based meatballs produced from this formulation deliver equivalent energy levels while offering a significantly higher total phenolic content (TPC) compared to traditional control formulas.
The texture of this plant-based product is a notable weakness, highlighting the need for further research to enhance it Adjustments to the formula, including the incorporation of transglutaminase enzyme and mycoproteins, could lead to improvements Additionally, exploring the development of new products from the waste of split gill mushrooms post-extraction may offer innovative solutions.
Recent studies on the split gill mushroom (Schizophyllum commune) have highlighted its phytochemical properties and potential health benefits Acanto et al (2022) conducted a comprehensive analysis, revealing significant cytotoxic activity alongside a detailed proximate analysis of the mushroom Additionally, Acanto and Cuaderes (2021) explored the antimicrobial properties of the ethanolic extract of Schizophyllum commune, further establishing its importance in scientific research These findings underscore the therapeutic potential of this mushroom, making it a subject of interest in multidisciplinary studies.
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APPENDIX A SENSORY RESULT OF PLANT-BASED MEATBALL
Table A.0.1 Sensory liking of the plant-based meatball with different split gill mushroom ratio
Measurements Control 5% SGM 10% SGM 15% SGM Color
Mean + SD with different lowercase superscripts in each column are significantly (p