Physicochemical, Microbiological, and Sensorial Properties of Chickpea Yogurt Analogue Produced with Different Types of Stabilizers

doi:10.21203/rs.3.rs-3241171/v1

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Physicochemical, Microbiological, and Sensorial Properties of Chickpea Yogurt Analogue Produced with Different Types of Stabilizers

https://doi.org/10.21203/rs.3.rs-3241171/v1

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There is a growing need for plant-based yogurts analogue that meet consumer demands in terms of texture and sensory qualities. Stabilizers are crucial in plant-based yogurt's physical properties which develop a thicker and creamier texture mimicking dairy yogurt. The addition of stabilizers helps to prevent syneresis. Thus, the study aims to evaluate the effect of pectin, corn starch, and locust bean gum (LBG) at different ratios on the physical, chemical, microbiological, and sensory properties of chickpea yogurts analogue (CYA). The concentration of stabilizer significantly influenced (p < 0.05) the proximate compositions, physicochemical and textural properties, and cell viability. A significant increase (p < 0.05) was observed in yogurt viscosity with the addition of corn starch and LBG at 1.0%. Firmness and consistency were improved in samples supplemented with 1.0% corn starch and commercial stabilizer. The sensory evaluation indicated that adding LBG at the ratio of 0.5% generated better preference among panelists in the appearance, color, and texture aspects despite commercial CYA showing significantly higher overall acceptability (p < 0.05) than other samples. The stabilizer's behavior significantly impacts the features of CYA which with 0.5% LBG received high consumer acceptance, which proves a good potential for CYA to be on the same shelf with other commercial yogurts analogue in the market.

Yogurt analogue

chickpea

stabilizers

physicochemical properties

sensory evaluation

Yogurt is one of the dairy products that is well-known for its nutritional values that improve the digestibility and bioavailability of some nutrients due to the acidification process and become a medium for fortification [1]. Due to the current trend where consumers are shifting to plant-based foods, the demand for plant-based yogurt analogue has also escalated. The term analogue refers to the resemblance of yogurt made from plant sources to traditional yogurt, particularly its appearance. The public is aiming to accomplish a healthy and sustainable diet [2] and the ethical issue that relates to the confining and slaughtering of animals [3]. The development of yogurts analogue is focused on consumers with health limitations which include cow milk protein allergy and lactose intolerance [4]. Yogurts analogue is also attracting the interest of vegetarians who are anticipating having wider food options that they can enjoy accomplishing their diet. Soymilk is common as a dairy milk substitute in the development of yogurts analogue owing to its high protein content and quality, and functional properties [5]. However, it also possesses off-flavors and allergens which makes it unappealing to consumers [6]. Therefore, other alternative legumes are being discovered as a potential ingredient in yogurt making such as chickpea (Cicer arietinum L.) as it is high protein content ranging from 18 to 29% [7, 8]. Chickpea also exhibits antioxidant, antibacterial, anticancer, and antidiabetic properties [9]. The production of yogurts analogue from chickpea (CYA) is predicted to have good functional properties with a lower cost of production. However, limited information is available on the suitability of the chickpea as a substrate in yogurt making, which requires further studies to be conducted.

As the yogurt quality is mainly influenced by textural and sensorial attributes, producing plant-based yogurt analogue with the ultimate body, texture, consistency, and free from syneresis remains the main technological challenge in the food industry [10]. Due to the nature of proteins, yogurt analogue tends to have structural disadvantages in contrast to yogurt made from cow milk [5]. Furthermore, plant proteins form poor gelation and network in an acidic environment which can result in syneresis at early storage or during storage [11]. Stabilizers are generally incorporated to overcome this drawback by enhancing the consistency and viscosity that resulted in lower whey separation, increased binding of free water, and maintenance of gel structure [12]. The stabilizers may increase the shelf life of the product and ensure uniformity among batches. The most suitable stabilizer for a specific yogurt analogue should be chosen as there are plenty of hydrocolloids available which perform varyingly in food systems [13]. Furthermore, the concentration at which the stabilizer is added to the yogurt formulation would affect its physical properties. Therefore, this study aims to evaluate the effect of different types of stabilizers (pectin, corn starch, and locust bean gum) at different ratios on the physical and chemical properties of chickpea yogurt analogue and to determine the consumer acceptance of chickpea yogurt analogue based on several sensory attributes.

2.1 Raw materials

Chickpea, sugar, vanilla powder, and corn starch were purchased from a local supermarket at Nilai, Negeri Sembilan. Pectin and locust bean gum (LBG) were purchased from Chemiz (Selangor, Malaysia) and Xintai Biological Technology Co. Ltd. (Shanghai, China), respectively. Yogurt starter culture was obtained from belle + bella (Lexington, MA).

2.2 Chickpea milk preparation

The process of extracting the milk was adapted with modifications on the ratio [14]. The chickpeas were soaked overnight and blended with distilled water at a ratio of 1:4 (chickpea to water; v: v) for 1 min. The blended legume was drained using a cotton cheesecloth to separate the extract from its solid residue. The chickpea milk was kept at refrigerated storage (4°C).

2.3 Yogurt preparation

The yogurt samples were prepared and the chickpea milk (1 L) was preheated to 40°C [14]. Sugar (3%) and vanilla powder (0.2%) were incorporated and homogenized using a heavy-duty mixer (L2R, Silverson Machines Ltd., Buckinghamshire, England) then, pasteurized at 80–85°C for 15 min and cool down to 42°C. The starter culture (0.5%) was inoculated, and the milk was incubated at 40°C for 16 h to obtain a set-type yogurt. The yogurt was stored at 5–8°C and used as a negative control. Stabilizers (pectin, corn starch and locust bean gum) at different ratios (0.5% and 1.0%) were added.

2.4 Proximate analysis

Moisture content, ash content, protein content, and crude fiber was determined according to the standard method of the Association of Official Analytical Chemists [15]. Fat content was determined using a Gerber method [16]. The carbohydrate content was calculated based on the overall proximate content.

2.5 pH, titratable acidity, and apparent viscosity

The pH value of the yogurt samples was determined at room temperature (24°C) using a pH meter (Delta 320, Mettler Toledo, Greifensee, Switzerland). The titratable acidity was acquired by direct titration method [17]. The viscosity of the yogurt was evaluated using a rotational rheometer (RheolabQC, Anton Paar, Graz, Austria).

2.6 Texture profile analysis (TPA)

The texture profile of yogurt (firmness, consistency, cohesiveness, and viscosity index) was analyzed using a texture analyzer (TA.HDPlus Connect, Stable Micro Systems Ltd., Surrey, UK) and employed a cylindrical probe with 36 mm diameter (P36 R). The compression test was done at pre-test and test speeds of 1 mm/s and a depth of 10 mm.

2.7 Viable cells count

The viability of the starter cultures in the yogurt samples was carried out after 1-week storage at 4°C. The sample (10 g) was homogenized in 90 mL sterile 0.1% peptone water (CM0009B, Oxoid, Ltd., Hampshire, UK), and proper serial dilutions were performed [18]. The diluted samples (1 mL) was inoculated on De Man, Rogosa, and Sharpe (MRS) agar (CM1153, Oxoid Ltd., Hampshire, UK) using the pour plate method and the agar plates were incubated in an anaerobic jar with the anaerobic bag at 37°C for 48 h. The total count of the viable cells was determined using a colony counter (Galaxy 230 Wiggens GmbH, Straubenhardt, Germany), and the results were calculated and expressed as Log₁₀ CFU/mL.

2.8 Sensory analysis

The sensory evaluation was conducted by employing an acceptance test method [19, 20] with 50 untrained panelists. A seven-points hedonic scale was employed.

2.9 Statistical analysis

All samples were analyzed in triplicate. The obtained data was analyzed using two-way Analysis of Variance (ANOVA) using Minitab 19.0 and Tukey’s test (Minitab, Pty Ltd., Sydney, Australia). The confidence level that was employed to decide the significant difference is 95% (p < 0.05). Data was expressed in mean with standard deviation (mean ± SD).

3.1 Proximate analysis

The proximate compositions (Table 1) show that different type of stabilizers significantly affects the moisture content ranging from 87.10–89.48%. For different concentrations, only LBG samples had no significant difference (p > 0.05) in moisture content, whereas pectin and corn starch samples displayed positive and negative correlations between the concentration of stabilizer and moisture content. This relates to the water retention ability of a particular stabilizer as it behaves diversely depending on the concentration [21].

Table 1

Proximate compositions of yogurts.
Sample	Moisture (%)	Fat (%)	Protein (%)	Ash (%)	Fiber (%)
Control A	89.48 ± 0.09^a	1.20 ± 0.00^cd	2.43 ± 0.01^Aa	0.24 ± 0.02^a	0.04 ± 0.01^c	6.87 ± 0.23^Ba
P (A)	87.84 ± 0.12^c	4.75 ± 0.07^a	2.39 ± 0.07^Aa	0.22 ± 0.03^ab	0.25 ± 0.02^c	5.40 ± 0.71^Ba
P (B)	89.58 ± 0.00^a	1.60 ± 0.00^bc	1.60 ± 0.10^Ab	0.18 ± 0.02^ab	5.24 ± 0.13^a	6.77 ± 0.47^Ba
CS (A)	88.50 ± 0.37^bc	1.45 ± 0.07^bc	2.20 ± 0.14^Aa	0.25 ± 0.01^a	0.51 ± 0.11^c	7.97 ± 0.60^Aa
CS (B)	87.10 ± 0.43^d	1.00 ± 0.00^c	1.66 ± 0.04^Ab	0.24 ± 0.01^a	3.36 ± 0.58^b	9.88 ± 0.59^Aa
LBG (A)	88.38 ± 0.19^bc	1.20 ± 0.00^de	1.70 ± 0.01^Ba	0.12 ± 0.03^b	0.12 ± 0.02^c	8.51 ± 0.14^Aa
LBG (B)	88.61 ± 0.28^b	1.70 ± 0.14^d	1.73 ± 0.12^Aa	0.23 ± 0.04^a	4.90 ± 0.54^a	8.08 ± 0.10^ABa
Note: Control A = without stabilizer, P(A) = pectin (0.5%), P(B) = pectin (1.0%), CS(A) = corn starch (0.5%), CS(B) = corn starch (1.0%), LBG(A) = locust bean gum (0.5%), LBG(B) = locust bean gum (1.0%).

For lowercase letters superscript; different letters are significantly different at p < 0.05. The same uppercase letters superscript in the same column are significantly different between the control and stabilizer variation for the same concentration (p < 0.05); means that do not share the same lowercase letters superscript in the same column are significantly different between the different concentrations within the same stabilizer (p < 0.05).

n.s. = not significant. Values are expressed as mean ± SD.

The fat content of the samples varied from 1.00–6.00%. According to CODEX Standard for fermented milk [22], dairy yogurt should possess milk fat less than 15%. The goal of the establishment of YA is to have low-fat content to address health concerns was achieved. There was a significant relationship (p < 0.05) between the stabilizers and concentrations of the fat content as a certain level of fat is required as it influences the texture, flavor, taste, and appearance of yogurt [23]. The protein contents recorded in this study for plant-based yogurt made from chickpea was better compared to the commercial ones made from hemp and coconut, but the values were rather similar to cashew- and almond-based yogurt [24]. The protein content was significantly reduced (p < 0.05) when the concentration of pectin and corn starch was elevated to 1.0%. The increment of stabilizer concentration induced a dilution effect, depending on the type of stabilizer [21]. Only LBG exhibits a significant effect (p > 0.05) on the ash content due to the high amount of sodium and calcium in LBG [25]. The results obtained for carbohydrate content (5.40–9.88%) were almost similar from the previous studies [23, 26–27]. For each type of stabilizer, the sample with 1.0% stabilizer showed significantly higher (p < 0.05) fiber content as compared to its respective sample with 0.5% stabilizer. The presence of fiber could be beneficial for the texture through an interaction with the casein matrix which resulted in better gel structure, and increased water-holding capacity and viscosity [28].

3.2 pH, titratable acidity and colour

The pH of yogurts containing pectin, corn starch, and LBG (Table 2) at both concentrations ranges from pH 4.00–4.22, while control A has the highest pH value of 4.33, which meets the standard of Food and Drug Administration (FDA) stating that yogurt should have pH of 4.6 or lower [29]. Increasing the concentration of stabilizer can cause pH reduction in yogurt due to the buffering capacity of the stabilizer, depending on its variety [21]. Based on the titratable acidity values, all samples possessed a lower percentage of titratable acidity as compared to the CODEX Standard for fermented milk which it specifies that yogurt should have a titratable acidity of at least 0.6% [30]. This may occur due to the absence of lactose as the plant substrate in the yogurt [24]. Therefore, the LAB strain used for specific plant-based milk should be critically determined to ensure an optimal fermentation is achieved [31].

Table 2

pH and titratable acidity values of different yogurt formulations.
Sample	pH	Titratable acidity (%)
Control A	4.33 ± 0.01^a	0.46 ± 0.02^bc
P (A)	4.11 ± 0.01^cd	0.59 ± 0.01^a
P (B)	4.14 ± 0.05^c	0.50 ± 0.02^b
CS (A)	4.00 ± 0.01^e	0.52 ± 0.01^b
CS (B)	4.07 ± 0.02^de	0.43 ± 0.01^c
LBG (A)	4.22 ± 0.03^b	0.51 ± 0.03^b
LBG (B)	4.22 ± 0.04^b	0.47 ± 0.07^bc
Note: Control A = without stabilizer, P(A) = pectin (0.5%), P(B) = pectin (1.0%), CS(A) = corn starch (0.5%), CS(B) = corn starch (1.0%), LBG(A) = locust bean gum (0.5%), LBG(B) = locust bean gum (1.0%). Means in the same column that do not share a letter (a,b,c) are significantly different (p < 0.05) for each sample

3.3 Textural properties

For the viscosity properties (Table 3), LBG(A) and LBG(B) samples showed significant differences (p < 0.05) due to the physical and chemical properties of the stabilizer itself [21]. LBG is composed of galactomannan with high molecular weight and low galactose residues as a side chain that contributes to its water-binding capacity, especially at high temperatures where it can completely solubilize [32]. LBG(A) showed a significantly higher viscosity as compared to control A, which indicates the ability of LBG to impart viscosity even at low concentrations [32]. P(A) and CS(B) samples also revealed an apparent viscosity higher than that of control A, attributed to the type and concentration of stabilizer used. A notable correlation between viscosity and fat content in their studies that involved dairy milk [21, 33]. The results acquired in the present study were in agreement with this since fat content in P(A) was higher than in P(B) which is dependent on the type of stabilizer. The high viscosity of the CS(B) sample is explained based on the ability of starch granules to absorb water and swell, which leads to restricted movement of the swollen granules [20].

The firmness of yogurts formulated with LBG at distinct concentrations showed no significant difference (p > 0.05) compared to pectin and corn starch, in which the values were significantly different (p < 0.05). The high viscosity of the LBG samples hinders the association of starter culture with other yogurt ingredients which prevents the yogurt to achieve a desired texture [34].

Table 3

Textural parameters of yogurts.
Sample	Apparent viscosity (mPa.s)	Firmness (g)	Consistency (g.sec)	Cohesiveness (g)	Viscosity index (g.sec)
Control A	185.35 ± 5.59^d	61.03 ± 1.21^cb	344.05 ± 0.60^c	-23.37 ± 0.25^b	-16.40 ± 1.56^b
P (A)	550.60 ± 2.97^b	128.44 ± 0.86^a	846.55 ± 16.68^b	-41.15 ± 0.25^c	-28.15 ± 1.34^c
P (B)	92.45 ± 1.20^e	35.61 ± 0.05^b	228.00 ± 2.11^e	-12.80 ± 0.21^a	-6.20 ± 0.08^a
CS (A)	145.95 ± 16.33^de	51.97 ± 0.05^b	291.13 ± 0.91^d	-9.75 ± 0.11^a	-4.34 ± 0.78^a
CS (B)	573.85 ± 37.12^b	138.68 ± 1.56^a	989.14 ± 0.99^a	-36.92 ± 1.31^c	-29.08 ± 0.66^c
LBG (A)	359.55 ± 12.80^c	45.89 ± 0.50^b	294.38 ± 0.11^d	-25.32 ± 0.11^b	-25.00 ± 0.66^c
LBG (B)	1460.50 ± 10.32^a	42.47 ± 1.41^b	237.17 ± 17.62^e	-39.48 ± 2.52^c	-46.30 ± 4.29^d
Note: Control A = without stabilizer, P(A) = pectin (0.5%), P(B) = pectin (1.0%), CS(A) = corn starch (0.5%), CS(B) = corn starch (1.0%), LBG(A) = locust bean gum (0.5%), LBG(B) = locust bean gum (1.0%). Means in the same column that do not share a letter (a,b,c,d,e) are significantly different (p < 0.05) for each sample

The yogurt is considered viscous and thick in consistency if it flows slowly on the spoon [31]. The inclusion of commercial stabilizer, pectin at 0.5%, and corn starch at 1.0% enhanced the consistency of the yogurt due to the increase in consistency as compared to control A. Next, cohesiveness determines the strength of internal bonds in the yogurt structure and is often associated with viscosity as well [34]. The cohesiveness of the sample improved only with corn starch at 0.5% and pectin at 1.0%. The increases in cohesiveness was accompanied by an elevated concentration of partially hydrolyzed guar gum supplemented in the set-type yogurt [30]. Based on this, it is assumed that the addition of a stabilizer helps in strengthening the molecular linkage in yogurt composition.

3.4 Viable cells count

The viability of lactic acid bacteria (LAB) in the samples was measured following one week of storage at 4°C and the findings were displayed in Table 4. A minimum of 10⁷ cfu/g of total starter culture is required in yogurt products [36]. The recent study showed that viable cells count for the samples ranged from log₁₀ 5.21 ± 0.12 CFU/mL to log₁₀ 7.13 ± 0.04 CFU/mL. It was shown that the addition of corn starch at higher concentrations boosted the cell viability in the yogurt whereas the cell viability was significantly decreased (p < 0.05) when the LBG level was higher. This was possible because the incorporation of corn starch at 1.0% imparted a favorable environment for the LAB to flourish. The reduction of viable cell count when the concentration of stabilizer was elevated from 0.5 to 1.0% [21] and depending on the concentration of stabilizer could provide optimum growth conditions for the starter cultures [21].

Table 4

Viable cells count of all yogurt formulations.
Sample	Viable cell count (log₁₀ CFU/mL)
Control A	6.70 ± 0.09^b
P (A)	5.45 ± 0.15^e
P (B)	5.21 ± 0.12^e
CS (A)	6.27 ± 0.06^c
CS (B)	7.13 ± 0.04^a
LBG (A)	6.63 ± 0.07^b
LBG (B)	5.77 ± 0.03^d
Note: Control A = without stabilizer, P(A) = pectin (0.5%), P(B) = pectin (1.0%), CS(A) = corn starch (0.5%), CS(B) = corn starch (1.0%), LBG(A) = locust bean gum (0.5%), LBG(B) = locust bean gum (1.0%). Means in the same column that do not share a letter (a,b,c,d,e) are significantly different (p < 0.05) for each sample

3.5 Sensory analysis

Sensory profile of the chickpea yogurt analogue was presented in Fig. 1. The commercial sample was a non-dairy oat milk yogurt and was used as a benchmark for the sensory profile of a plant-based yogurt analogue (YA) product. The results showed that control A has the lowest rating for all attributes with appearance and texture receiving significantly lower (p < 0.05) scores as opposed to other samples. This may be resulting from the absence of any stabilizer in the yogurt that made it unappealing to the panelists [19, 35].

Furthermore, CY showed no significant difference (p > 0.05) with the YA sample for appearance, color, and texture except for flavor. The developed CY formulation could mimic the commercially available YA in terms of these important sensory attributes. CY samples were rated significantly higher (p < 0.05) than other samples in the aspect of flavor, aftertaste, and overall acceptability. This was apparently due to the taste of oat that was more acceptable, given that oat-based foods are more popular and highly consumed as opposed to chickpea-based food.

To conclude, the employment of different stabilizers at different ratios significantly impacted the physical and chemical properties of chickpea yogurt analogue. The effects of stabilizers were obvious on pH value, syneresis, cell viability, textural and sensory quality of the yogurt. This was nearly similar for the concentrations of stabilizer. The suitable concentration of stabilizer is employed in specific YA must be determined to obtain an optimum physical and chemical characteristic.

Ethics approval and Consent to Participate: Not applicable.

Consent for publication: The results/data/figures in this manuscript have not published elsewhere, nor they are under consideration by another publisher.

Conflicts of interest: The authors have no conflicts of interests (financial or non-financial) to declare that are relevant to the content of this article.

Authors’ contribution statements: Authors 1 (S.N.M.F), 2 (A.A.M), and 6 (A.Z.M) wrote the manuscript test. Authors 3 (A.S.M.H), 4 (M.H.A.R), 5 (I.N.M), and 6 (A.Z.M) provide supervision, research materials, and funding acquisition. All authors reviewed the manuscript.

Funding: This work was supported by the Universiti Putra Malaysia (Grant numbers: GP-IPM/2022/9740300).

Data availability: All dataset used in this study has been provided in this article.

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Editorial decision: Major revision
31 Aug, 2023
Editor assigned by journal
29 Aug, 2023
Submission checks completed at journal
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Physicochemical, Microbiological, and Sensorial Properties of Chickpea Yogurt Analogue Produced with Different Types of Stabilizers

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Version 1

Abstract

Figures

1. Introduction

2. Materials and Methods

2.1 Raw materials

2.2 Chickpea milk preparation

2.3 Yogurt preparation

2.4 Proximate analysis

2.5 pH, titratable acidity, and apparent viscosity

2.6 Texture profile analysis (TPA)

2.7 Viable cells count

2.8 Sensory analysis

2.9 Statistical analysis

3. Results and Discussion

3.1 Proximate analysis

3.2 pH, titratable acidity and colour

3.3 Textural properties

3.4 Viable cells count

3.5 Sensory analysis

4. Conclusion

Declarations

References

Additional Declarations

Status:

Version 1