TECNOCIENCIA CHIHUAHUA, Vol. XVIII (4): e1614 (2024)

https://vocero.uach.mx/index.php/tecnociencia

ISSN-e: 2683-3360

Research Article

Physicochemical and functional properties of yogurt

enriched with ultrafiltered juice of Red Prickly Pear

(Opuntia ficus-indica) as a functional ingredient

Propiedades fisicoquímicas y funcionales de yogurt enriquecido con

jugo ultrafiltrado de tuna roja (Opuntia ficus-indica) como ingrediente

funcional

*Correspondencia: sugey.sinagawagr@uanl.edu.mx (Sugey R. Sinagawa-Garcia)

DOI: https://doi.org/10.54167/tch.v18i4.1614

Recibido: 16 de agosto de 2024; Aceptado: 13 de noviembre de 2024

Publicado por la Universidad Autónoma de Chihuahua, a través de la Dirección de Investigación y Posgrado.

Editor de Sección: Dr. Armando Quintero-Ramos

Abstract

Yogurt has long been recognized as a health-promoting food for its bioavailability of protein, energy,

probiotics and calcium, increasing digestibility, as well as preventing potential diseases beyond basic

nutritional function. Recently, yogurt has become one of those foods whose functional properties are

developed by adding functional substances with a potential positive effect on health. The objective

was to evaluate the effect of ultrafiltered juice of red fruits of O. ficus-indica as a functional ingredient

in yogurt. A completely randomized block design (CRBD) was used. The treatments were: yogurt

with 0 % ultrafiltered red prickly pear juice (Y0) as a control; yogurt with 10 % ultrafiltered red

prickly pear juice (Y1); yogurt with 20 % ultrafiltered red prickly pear (Y2); and yogurt with 30 %

ultrafiltered red prickly pear juice (Y3). The variables to measure in each yogurt treatment were pH

during lactic fermentation and the final product, syneresis, energy content, colorimetry, composition

(total solids, protein, fat and ash), antioxidant activity (DPPH, ABTS and FRAP), phenol content and

sensory evaluation. The incorporation of red prickly pear ultrafiltrates accelerated antioxidant

activity and total phenolic contents. Finally, the sensory panel satisfactorily accepted the

incorporation of red prickly pear ultrafiltered in yogurt.

Alondra Valadez-Pineda1, Gerardo Méndez-Zamora1, Nydia Vásquez-Aguilar1, Luisaldo

Sandate Flores2, Carlos Alberto Hernández-Martínez1 and Sugey R. Sinagawa- García1*

1 Universidad Autónoma de Nuevo León, Facultad de Agronomía, UANL. Av. Francisco Villa S/N, Col. Ex

Hacienda el Canadá, General Escobedo, Nuevo León, México. C.P. 66050

2Tecnológico Nacional de México/ITS de Rioverde. Carretera Rioverde-San Ciro Km 4.5, Rioverde, San

Luis Potosí

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Keywords: ultrafiltered, yogurt, antioxidant activity, phenolic content, sensory panel.

Resumen

El yogurt ha sido reconocido desde hace mucho tiempo como un alimento que promueve la salud

por su biodisponibilidad de proteínas, energía, probióticos y calcio, aumentando la digestibilidad,

además de prevenir enfermedades potenciales más allá de la función nutricional básica.

Recientemente, el yogurt se ha convertido en uno de los alimentos cuyas propiedades funcionales se

desarrollan mediante la adición de sustancias funcionales con un efecto potencial positivo en la salud.

El objetivo fue evaluar el efecto de jugo ultrafiltrado de frutos rojos de O. ficus-indica como ingrediente

funcional en yogurt. Se utilizó un diseño de bloques completamente al azar (DBCA). Los tratamientos

fueron: yogurt control 0 % jugo ultrafiltrado de tuna roja (Y0); yogurt con 10 % de jugo ultrafiltrado

de tuna roja (Y1); yogurt con 20 % de jugo ultrafiltrado de tuna roja (Y2); y yogurt con 30 % de jugo

ultrafiltrado de tuna roja (Y3). Las variables para medir en los tratamientos de yogurt fueron pH

durante la fermentación láctica y en el producto final, sinéresis, contenido energético, colorimetría,

composición (sólidos totales, proteína, grasa y ceniza), actividad antioxidante (DPPH, ABTS y FRAP),

contenido de fenoles y evaluación sensorial. La incorporación de ultrafiltrados de tuna roja aceleró

la fermentación del yogurt además se incrementó la sinéresis, el contenido de energía y pH. Los

ultrafiltrados de tuna roja en yogurt aumentaron la actividad antioxidante y el contenido de fenoles

totales. Finalmente, el panel sensorial acepto satisfactoriamente la incorporación de ultrafiltrados de

tuna roja en yogurt.

Palabras clave: ultrafiltrado, yogurt, actividad antioxidante, contenido fenólico, panel sensorial.

1. Introduction

Nowadays, the consumption of functional foods is in high demand because consumers

consider that their health is linked with the foods they ingest (Dinkçi et al., 2021). Foods or

ingredients classified as functional foods resemble traditional ones but may have the potential to

provide additional health benefits beyond the standard nutrient content, and should be incorporated

as part of a normal diet for consumption in a local setting or specific culture (Shori et al., 2022).

Functional foods require new bioactive ingredients which can be used by the food industry (Faustino

et al., 2019). In the past years, studies about horticultural crops have revealed that these products

have important functions in human health due to high levels of active biological compounds (Settar

Unal et al., 2022).

Cacti are a group species that live in arid areas; México hosts 518 species out of which 47.7 % are

endemic (Martínez et al., 2021). Opuntia ficus-indica (L.) Mill (Cactaceae) is a plant considered as

functional food for its nutritional benefits on human health due to its high content of antioxidant

compounds in its cladodes, flowers and fruits (Ayala et al., 2021). The fruits of O. ficus-indica are

berry-like and are known as “tunas” in Mexico, they contain flavonoids, phenolic acids, betalains,

ascorbic acid, fatty acids, lignans and sterols (Martínez et al., 2021). Betalains are derived from

betalamic acid and are divided into two structural subgroups: betacianines and betaxantines. These

compounds are extracted commercially from beetroot and are utilized commonly in the food

industry as a color additive in many types of foods, such as dairy products, candies, soft drinks and

some emulsified meat products (Bassama et al., 2020). Many studies have investigated the properties

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and interactions of the pigments of prickly pear (betalains), for example, characterizing and

quantifying betalains and phenolic compounds (Gómez-Maqueo et al., 2020); evaluating the

antioxidant potential, antiinflamatory and anti hyperglucemic activity of the extracts of O. ficus-

indica (Gómez-Maqueo et al., 2019); and studying the potential of semi-processed juices from O. ficus-

indica as a natural antimicrobial against Gram-negative and Gram-positive food pathogenic bacteria

(Palmeri et al., 2020).

According to the lastest studies, the use of numerous additives, fruits or bioactive compounds of

plant origin in the production of yogurt has a significant improvement in its quality and benefits

such as antioxidant properties (Cenobio et al., 2019; Tavakoli et al., 2019; Won-Young et al., 2020).

Yogurt is classified as a functional food and it is one of the most consumed dairy products (Jeong et

al., 2018). It is produced by the coagulation of proteins during fermentation with lactic acid bacteria

such as Lactobacillus bulgaricus and Streptococcus thermophilus under defined time and temperature

(Kim et al., 2019; Cho et al., 2020). These bacteria ferment lactose, producing lactic acid, carbon

dioxide, acetic acid, dactyls, acetaldehyde and other components that give yogurt its characteristic,

flavor and smell; and they must be viable and abundant in the final product (Shori et al., 2022). Yogurt

is a source of bioactive compounds that are formed during lactic fermentation and has a limited

content of antioxidant activity. For this reason, the production of yogurt enriched with antioxidants

from natural sources is of considerable interest and presents a novel approach for product

development (Dabija et al., 2018). In summary, yogurt and red prickly pear are both functional foods

that, when combined, may have physicochemical and functional properties that are beneficial to

human health. Therefore, the objective is to evaluate the effect of ultrafiltered juice of red fruits of O.

ficus-indica as a functional ingredient in yogurt.

2. Materials and methods

2.1 Materials

For the yogurt production, commercially available ultra-pasteurized milk, sugar, bacterial cultures,

and ultrafiltered juice from red prickly pear (O. ficus-indica) were used, as shown in Table 1.

Table 1. Yogurt treatments with ultrafiltered red prickly pear juice (O. ficus-indica) as a functional ingredient.

Tabla 1. Tratamientos del yogurt con jugo ultrafiltrado de tuna roja (O. ficus-indica) como ingrediente funcional.

Ingredient (%) †

Treatment ‡

Ultrapasteurized milk

90.90

90.49

90.08

89.68

Saccharose

9.09

9.05

9.01

8.97

Bacteria cultures

0.01

PUJ O. ficus-indica

0.00

0.45

0.90

1.35

†PUJ: Pasteurized ultrafiltered juice. ‡ Y0: control yogurt; Y1: yogurt with 10 % ultrafiltered red prickly pear juice;

Y2: yogurt with 20 % ultrafiltered red prickly pear juice; Y3: yogurt with 30 % ultrafiltered red prickly pear juice.

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2.2 Methods

2.2.1 Yogurt processing

The elaborate process of yogurt formulation (Table 1) was done following the Karnopp

method et al., (2017) with some modifications. Saccharose was added to the ultra pasteurized milk

and the mixture was heated to 42 °C. At this temperature, the freeze-dried bacterial cultures and

pasteurized ultrafiltrate of red prickly pear juice were added to each treatment. The mixture was

then incubated at 42 °C for 6 h. Subsequently, the products were cooled to 7 °C and kept at that

temperature for 12 h for the analysis of the variables. The commercial ultra-pasteurized milk used in

the experiment was composed of 33.1 % total solids, 3.1 % protein, 2.9 % fat and 0.6 % ash.

2.2.2 Physicochemical analysis

The pH of the yogurt samples was measured with a potentiometer (EcoSense pH1000A, YSI,

Ohio, USA) during fermentation and in final products.

Yogurt syneresis was performed according to the method of Jeong et al. (2018). Samples of 10 g of

yogurt were analyzed, which were centrifuged at 600 × g for 6 min at 4 °C. The supernatant (serum)

obtained was weighed, and the syneresis was expressed as a percentage weight of the serum

separated from the initial sample (Kim et.al., 2020) using the following Eq. (1):

󰇛󰇜 󰇣 󰇛󰇜

󰇛󰇜󰇤   Eq. (1)

Gross energy was determined using an adiabatic bomb calorimeter (Bomb Calorimeter, Parr

Instrument, Illinois, USA), using petroleum ether as the combustion reagent.

2.2.3 Colorimetry

The color was determined by the values of CIE L* (0 = black; 100 = white), CIE a* (- green; +

red), CIE b* (- blue; + yellow), Chroma (C* = intensity of the color) and Hue angle (hue), with a

colorimeter (CR 410, Konica Minolta, Japan). Furthermore, the color difference (∆E) between the

color of the samples (L2) compared to the control (L1), was calculated according to Dinkçi et al. (2021)

with the following Eq. (2):

  󰇛 󰇜 󰇛 󰇜 󰇛 󰇜 Eq. (2)

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2.2.4 Proximal composition

The total solids content was determined according to the method 930.15 (AOAC, 2005) by

gravimetric determination of the moisture loss of the sample dried for 3 h in an oven (Yamato

Scientific America Inc. Constant Temperature Oven, Japan) at 103 °C. The percentage of ash

according to the method 942.05 (AOAC, 2005) was considered as the residue after incineration of the

samples at 550 °C in a muffle (Thermo Scientific, Waltham, MA) for 4 h. The percentage of protein

was determined by the Kjeldahl method described by Wijesekara et al., (2022). In a digestion block,

the samples were digested at 400 °C with sulfuric acid and catalyst (3 % CuSO4, 97 % K2SO4).

Subsequently, the samples were subjected to a Kjeldahl distiller with NaOH to distill the nitrogen

from the combustion, which once obtained was titrated with hydrochloric acid (0.1 N). The results

obtained were the quantification of total nitrogen in the samples, and through a conversion factor.

Fat quantification by method 920.39 (AOAC, 2005) consisted of quantifying the ether extract obtained

by extracting triacylglycerides from the samples, using ether in an adiabatic calorimetric fat extractor

(Parr Instrument, USA).

2.2.5 Preparation of yogurt-water extracts

The preparation of the yogurt-water extracts was done according to the method of Shori et

al. (2022). A total of 40 g of each yogurt sample was homogenized with 10 mL of dH2O. The pH of the

samples had to be 4.0, then the samples were incubated in a water bath at 45 °C for 10 min. The

samples were centrifuged at 5000 rpm at 4 °C for 10 min, and the supernatant was adjusted to pH 7.0

using NaOH (0.1 M). A second centrifugation was carried out (5000 rpm, 4 °C, 10 min) and the clear

supernatant was used for subsequent analyses: DPPH, ABTS+, FRAP and phenolic content.

2.2.6 Antioxidant activity

The DPPH (2,2-diphenyl-1-picrylhydrazyl) assay was based on what was proposed by Jeong

et al., (2018) with some modifications. The DPPH solution was dissolved in 80 % methanol, then the

reagent was diluted 1:10 to an absorbance of 0.700 at 515 nm. The previously prepared yogurt-water

extract (25 µL) was mixed with the DPPH solution (975 µL), and incubated in the dark at room

temperature for 30 min. A mixture of 25 µL of ethanol and 975 µL of DPPH solution was used as a

control. The antioxidant activity was calculated in accordance with Jeong et. al. (2018) as Eq. (3):

 󰇛󰇜 󰇣  󰇡

󰇢󰇤   Eq. (3)

The ABTS (2,2'–azino–bis–(3–ethylbenzothiazolin–6–sulfonic acid) assay was performed based on

Jeong et al., (2018) with modifications. The ABTS reagent (14.8 mM) was mixed with 5 mM potassium

sulfate, and the sample was incubated in the dark at room temperature for 16 hours. The working

solution (ABTS) was diluted with dH2O to an absorbance of 0.700 ± 0.05 at 734 before use. The yogurt-

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water extract samples (10 µL) were mixed with 1 mL of ABTS solution and incubated in the dark at

room temperature for 15 min. The percentage of antioxidant activity was calculated according to

Jeong et al. (2018) in the following Eq. (4):

󰇛󰇜 󰇣  󰇡

󰇢󰇤   Eq. (4)

The methodology for FRAP was performed according to Schneider et al., (2022) with some

modifications. The solution was prepared by mixing acetate buffer (300 mM, pH 3.6) with 10 mM

TPTZ solution (2,4,6-tripyridyl-s-triazine) in 40 mM HCl and 20 mM FeCl3, in a ratio of 10: 1:1 (v/v/v).

The mixture of solutions was heated to 37 °C and 25 μL of diluted sample (1/5) or Trolox standard

solution was added to 1 mL of FRAP solution. The absorbance was measured at 593 nm, after 30 min.

The equation curve was based on Trolox concentrations between 0 and 1200 µM, and the results were

expressed in µmol TE/L.

The Folin-Ciocalteu method (Wijesekara et al., 2022) with some modifications was used to quantify

the total phenolic content in the yogurt-water extracts. The analysis was based on a nine-point

calibration curve from 0 to 0.032 mg/mL with intervals of 0.004 mg/mL gallic acid. The absorbance

was measured at 760 nm before use. The results were expressed in gallic acid equivalents (mg AG/L).

2.2.7 Sensory evaluation

The sensory evaluation was carried out based on Shori (2020) with some modifications. The

yogurt samples were evaluated on day 1 of storage. Sample preparation consisted of placing 20 g of

each treatment in plastic containers, assigning them a random three-digit number. The panel

consisted of 40 people (were students and professors, all yogurt consumers) and was divided into

two sections. In the first section, 20 panelists evaluated the four treatments of replica one, while the

other 20 assessed the same treatments in replica two. This way, the evaluation was conducted in

replicates, ensuring that no different treatments were used, which allowed for more consistent and

comparable results. The evaluation was carried out with a 5-point hedonic scale (5= I like it a lot, 4= I

like it, 3= I neither like it nor dislike it, 2= I dislike it, 1= I dislike it very much), for the five attributes:

color (presence of pink), milky odor, flavor, viscosity and overall acceptability.

2.2.8 Statistical analysis

The variables analyzed in the filtered, retained and pasteurized samples were considered as

a treatment effect (Ƭi) to test the hypothesis H0 (equality of treatments - no effect on the variables);

this analysis was based on the general linear model “yij=μ+τi+eij” where "yij" refers to the study

variable, "μ" refers to the global mean, "Ƭi" refers to the effect of the i treatment and "eij" is the

experimental error.

In the yogurt experiment, a factorial design was first carried out to test the hypothesis (H0) about the

difference in pH between the treatments (Ƭi) in relation to the time of lactic fermentation: yij=μ+τi+tj+

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(τi)ij(k)+β+eij; yij= study variable (pH); µ= overall mean; Ƭi=effect of the ith treatment; tj= effect of the

jth fermentation time; (Ƭi)ij(k)= effect of the treatment-fermentation time interaction; B= block effect; eij

= experimental error.

In the treatments (Ƭi) of the final yogurt product, the hypothesis H0 was tested based on:

yij=μ+τi+βj+eij; yij= study variable; µ= overall mean; Ƭi=effect of the ith treatment; Bj= effect of the ith

block; eij = experimental error.

A value less than 0.05 (P-Value) was the criterion to reject H0. The rejected variables were evaluated

by a comparison of means (Tukey). Minitab® 17.1.0 (2013) software was used for data analysis.

Measurements of the colorimetry, pH and soluble solids variables were carried out seven times.

Meanwhile, measurements of pH variables during lactic fermentation, antioxidant activity, betalains,

phenolic content, total sugars, reducing sugars, proximal composition and water retention capacity

were carried out in triplicate.

3. Results and discussion

3.1 Lactic fermentation

During lactic fermentation, the interaction of treatments and time showed no effect (P>0.05)

for pH. Thus, the behavior of pH over hours is shown in Table 2. The pH was different (P<0.05)

between the treatments at 0, 2 and 6 h. The yogurt with the highest content (Y3) of ultrafiltrates

showed lower pH at hours 0 and 2, given that the pH of the pasteurized ultrafiltrate was 4.57.

Table 2. pH behavior during yogurt fermentation

Tabla 2. Comportamiento del pH durante la fermentación del yogurt

Treatment†

Time (h)

6.568a

6.013a

4.637

4.117ª

6.490a

5.913b

4.487

4.062b

6.382b

5.868b

4.502

4.053b

6.295b

5.783c

4.583

4.092ab

SEM

0.034

0.102

0.056

0.028

P-Value

0.000

0.059

0.044

† Y0: control yogurt; Y1: yogurt with 10 % ultrafiltered red prickly pear juice; Y2: yogurt with 20 %

ultrafiltered red prickly pear juice; Y3: yogurt with 30 % ultrafiltered red prickly pear juice. SEM:

standard error of the mean. a-c Means in columns with different letters differ statistically (P<0.05).

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Similar results were reported by Jeong et al. (2018), who used green tea powder in the yogurt

formulation, where the pH was lower. Those authors indicated that the addition of phenolic

compounds and organic acids can improve the metabolic activity of lactic acid bacteria in yogurt,

leading to faster acidity of milk and, therefore, a shortening of fermentation times.

3.2 Physicochemical properties

Table 3 reports the physicochemical properties of the yogurt, such as pH, syneresis and

energy. These properties showed a difference (P>0.05) between yogurt samples. Y0 presented the

lowest syneresis value, and Y3 the highest energy value.

Table 3. Physicochemical properties of yogurt with ultrafiltered red prickly pear juice.

Tabla 3. Propiedades fisicoquímicas del yogurt con jugo ultrafiltrado de tuna roja.

Variables

Treatment†

SEM‡

P-Value

4.01a

3.99ab

3.97b

4.01a

0.01

0.047

Syneresis %

60.73c

65.50b

68.56ab

71.36a

1.09

0.000

Energy (Kcal/kg)

4285.50b

4164.50bc

4061.00c

4567.50a

19.10

0.001

† Y0: control yogurt; Y1: yogurt with 10 % ultrafiltered red prickly pear juice; Y2: yogurt with 20 %

ultrafiltered red prickly pear juice; Y3: yogurt with 30 % ultrafiltered red prickly pear juice. ‡ SEM:

standard error of the mean. a-c Means in rows with different letters differ statistically (P<0.05).

The pigmentation of betalains is most stable in a pH range of 3 to 7, with betacyanins being more

resistant to acidic conditions and betaxanthins being more stable at neutral pH (Schneider et al.,

2022). The addition of red prickly pear ultrafiltrates differed in pH value, Y0 and Y3 presented the

highest value. The results obtained present more acidic values compared to previous research in

which leaf extracts were added; For example, the addition of aqueous extracts of herbs in yogurt

presented pH of 4.62 to 4.36 on day 1 of storage (Dabija et al., 2018), and the addition of lotus leaf

powder showed values of 4.39 and control of 4.49 (Da-Hee et al., 2019).

Syneresis values differed significantly between treatments (P<0.05). Y0 presented the lowest

percentage in terms of serum release compared to treatments added with red prickly pear

ultrafiltrates. Similar results were obtained by Wijesekara et al., (2022), who added aqueous natural

colorants to yogurt after fermentation, compared to this study in which the ultrafiltrates were added

before fermentation, so adding the aqueous extracts before or after fermentation generates the same

effect in the release of serum. T

The variation in the energy content of yogurt is due to the interpretation of the carbohydrate, protein

and fat contents in the formulations (Ahmed et al., 2023). Y3 presented the highest energy content,

followed by Y0, Y1 and Y2. However, it would have been expected that the higher the concentration

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of red tuna ultrafiltrate, the higher the energy content, but this was not the case for Y2, perhaps due

to variations in the formulation.

3.3 Colorimetry

The color parameters evaluated in yogurt showed significant differences (Table 4). L*, b* and

hue (Hue angle) decreased as the amount of ultrafiltrates added increased, while a* and saturation

(Chroma) increased.

Table 4. Colorimetry in yogurt with ultrafiltered red prickly pear juice.

Tabla 4. Colorimetría en yogurt con jugos ultrafiltrados de tuna roja.

Colorimetry†

Treatment‡

SEM§

P-Value

93.81a

79.63b

73.51c

69.66d

0.91

0.000

-2.72c

18.68b

25.59a

29.46a

1.24

0.000

9.41a

3.89b

2.98b

2.87b

0.44

0.000

Chroma

9.79c

19.22b

25.86a

29.7a

1.09

0.000

Hue Angle

106.20a

13.53b

7.86c

6.66c

0.99

0.000

∆E

Ref

19.55c

27.45b

32.19a

0.11

0.000

† a*: red coordinates; b*: yellow coordinates; L*: luminosity; ∆E: color difference, Y0 as reference. ‡ Y0:

control yogurt; Y1: yogurt with 10 % ultrafiltered red tuna juice; Y2: yogurt with 20 % ultrafiltered

red tuna juice; Y3: yogurt with 30 % ultrafiltered red tuna juice. § SEM: standard error of the mean. a-

d Means in rows with different letters differ statistically (P<0.05).

Cenobio et al. (2019) reported that L* increases in proportion to the concentration of the purple O.

ficus-indica betalain emulsion. Also, the value of a* increases, since the emulsions tend to color red

because of the presence of betalains. Parameter b* increases during the shelf life of the yogurt. In the

results obtained, a* increased when adding 20 and 30 % of red tuna ultrafiltrates.

Various types of acids and their quantities cause different effects on the color of betalains (Guneser,

2021); an example is lactic acid produced during milk fermentation. Guneser (2021) described a

relationship: as the shelf life of yogurt enriched with beet extract progressed, the lactic acid content,

pH, and the value of L* and b* increased, while the value of a* decreased. In this case, the pH value

decreased as the percentage of ultrafiltered red tuna juice increased and the a* value increased, so

the use of betalains from red tuna is more stable at acidic pH, by obtaining greater pigmentation in

red coordinates.

In addition, Guneser (2021) reported a significant positive correlation between Chroma values and

betalain content. A similar effect occurred in the treatments, Chroma and a* increased as the

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percentage of ultrafiltrates increased. The color difference between the samples results when 3.5 <

ΔE < 5.0 (Schneider et al., 2022), in treatments Y1, Y2 and Y3 the value was greater than 5.0.

3.4 Proximal composition

The results of the proximal composition such as total solids, fat and ash did not present

significant differences (P>0.05), but the levels of proteins differed significantly (P<0.05) between

treatments, with Y2 and Y3 having lower content (Table 5).

Table 5. Proximal composition of yogurt with ultrafiltrates of red prickly pear juice.

Tabla 5. Composición proximal en yogurt con ultrafiltrados de tuna roja.

Composition (%)

Treatment†

SEM‡

P-Value

Total solids

19.15

18.25

17.50

16.85

0.541

0.173

Raw protein

2.15ab

2.50a

1.75b

1.65b

0.075

0.011

Raw fat

0.90

1.10

0.65

0.125

0.201

Ash

0.65

0.60

0.55

0.029

0.292

† Y0: control yogurt; Y1: yogurt with 10 % ultrafiltered red tuna juice; Y2: yogurt with 20 %

ultrafiltered red tuna juice; Y3: yogurt with 30 % ultrafiltered red tuna juice. ‡ SEM: standard error of

the mean. a-b Means in rows with different letters differ statistically (P<0.05).

The chemical composition of yogurt depends on the composition of the milk, the presence of

additives and processes (Guneser, 2021). The fat and protein values of the four yogurt treatments are

within the range established by NOM-181-SCFI/SAGARPA-2018, since the minimum percentage of

protein must be at least 1.60 and the maximum percentage of butterfat must be 7.0. Although the

percentage of protein decreased in the yogurt treatments as the amount of ultrafiltrates increased.

Guneser (2021) reported 13.13 g/100 g of dry matter, 3.01 g/100 g of fat, 2.50 g/100 g of protein and

0.85 g/100 g of ash, in yogurt with cow's milk. In the results obtained from Y0, Y1, Y2 and Y3, the fat,

protein and ash contents were lower.

3.5 Antioxidant activity and total phenolic content

The antioxidant activity and total phenolic content are shown in Table 6. The ultrafiltrates

increased the results obtained from DPPH, ABTS, FRAP and total phenolic content in Y1, Y2 and Y3,

with Y3 presenting the highest values.

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Table 6. Antioxidant activity and total phenolic content in yogurt with ultrafiltrates of O. ficus-indica red variety.

Table 6. Actividad antioxidante y contenido de fenólicos totales en yogurt con ultrafiltrados de O. ficus-indica

variedad roja.

Variable†

Treatment‡

SEM§

P-Value

DPPH• %

2.69c

15.86b

27.02a

33.21a

1.97

0.000

ABTS•+ %

15.50d

19.25c

22.91b

28.34a

0.75

0.000

FRAP (µmol TE/L)

0.167c

0.239b

0.310ab

0.356a

0.018

0.000

CFT (mg AG/L)

22.74c

34.69b

39.68ab

44.61a

2.15

0.000

† TE: Trolox equivalents; TPC: Total phenolic content; FA: gallic acid. ‡ Y0: control yogurt; Y1: yogurt

with 10% ultrafiltered red tuna juice; Y2: yogurt with 20% ultrafiltered red tuna juice; Y3: yogurt

with 30% ultrafiltered red tuna juice. § SEM: standard error of the mean. a-d Means in rows with

different letters differ statistically (P<0.05).

The antioxidant activity reported by different methodologies such as DPPH, ABTS and FRAP, as

well as the total phenolic content showed a similar behavior between the treatments, given that the

antioxidant activity is correlated with the phenolic content (Dabija et al., 2018); because as the

percentage of red tuna ultrafiltrates increased in the treatments, the percentage of radical inhibition

by DPPH, ABTS, and the content of µmol TE/L in FRAP increased, and in turn, the content of total

phenols increased, with Y3 reporting the highest results.

In yogurt enriched with microencapsulated fruits of purple O. ficus-indica, similar results were

obtained to those reported in the DPPH and ABTS assay (Cenobio-Galindo et al., 2019).

Schneider et al. (2022) mentioned that the betacyanin-rich fraction has a higher antioxidant activity

and phenolic content than the betaxanthin-rich fraction. The ultrafiltered red tuna juice added to the

different yogurt treatments presented a higher content of betacyanins (93.74 mg/L) than betaxanthins

(39.18 mg/L), resulting in the increase in antioxidant activity and CFT in Y1, Y2 and Y3.

3.6 Sensorial evaluation

Table 7 shows the results obtained from the sensory evaluation of yogurt. The evaluation

was conducted in replicates, using the same treatments, which allowed for data grouping. The pink

color and flavor showed significant differences (P<0.05) between the treatments, as well as the milky

odor, viscosity and overall acceptability (AG).

Table 7. Sensorial evaluation of yogurt with ultrafiltrates of O. ficus-indica juice red variety.

Tabla 7. Evaluación sensorial en yogurt con ultrafiltrados de O. ficus-indica variedad roja.

Treatment†

Pink color

Milky odor

Flavor

Viscosity

1.750c

4.000a

4.250a

4.000a

4.000ab

4.000a

3.750b

4.000a

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4.250a

4.000a

4.000ab

4.000a

4.000ab

4.000a

4.000ab

4.000a

P-Value

0.000

0.084

0.009

0.430

0.399

† Y0: control yogurt; Y1: yogurt with 10 % ultrafiltered red tuna juice; Y2: yogurt with 20 %

ultrafiltered red tuna juice; Y3: yogurt with 30 % ultrafiltered red tuna juice. OA: Overall

acceptability. a-b Means in columns with different letters differ statistically (P<0.05).

The yogurt treatments enriched with ultrafiltered red tuna juice (Y1, Y2, and Y3) received sensory

scores similar to those of the control yogurt (Y0), without any negative effects. Wijesekara et al. (2022)

reported that the use of natural pigments in yogurt production does not negatively impact sensory

perception. However, in this study, the inclusion of red tuna ultrafiltrates resulted in a noticeable

pink color and a distinct flavor profile. While Y0 did not exhibit pink pigmentation and was the

panelists' accepted treatment in terms of flavor, Y2 and Y3 were also well-received, though their

flavor scores were slightly lower than Y0.

4. Conclusions

The incorporation of red tuna ultrafiltrates into yogurt resulted in several significant effects

on its properties. First, there was an acceleration in the yogurt fermentation process. Additionally,

the inclusion of these ultrafiltrates increased syneresis, energy content, and pH, particularly in the

Y3 sample. However, a decrease in L*, b*, and Hue angle values was also observed, indicating a

noticeable color difference compared to yogurt without additives (Y0). Furthermore, the addition of

ultrafiltrates reduced protein content in the Y2 and Y3 yogurt variants. From a health perspective,

the presence of red tuna ultrafiltrates significantly boosted antioxidant activity (measured through

the DPPH, ABTS, and FRAP methods) and increased total phenolic content. A sensory panel

evaluated the yogurt with the ultrafiltrates positively, suggesting favorable consumer acceptance.

This sensory evaluation was conducted on a pilot scale, so future research should include a larger

group of consumers to obtain more reproducible and reliable results. The treatments containing 20%

and 30% ultrafiltered tuna juice were the best-rated percentages by the panelists. Ultrafiltered red

tuna appears to be a beneficial addition to yogurt, enhancing certain nutritional and functional

aspects, though it also brings changes to the yogurt's physical and protein properties that warrant

further consideration. Further large-scale studies involving a more substantial number of sensory

panelists are recommended to gain more robust and representative data on consumer acceptance.

Additionally, investigating the product's stability during storage—evaluating whether the

antioxidant benefits and changes in physical properties are maintained over time—along with

determining the product's shelf life under commercial conditions, would provide valuable insights.

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Acknowledgments

Many thanks to Saba Yasin MSc for his thorough review and insightful editing of this

manuscript, greatly enhancing its quality.

Author’s contribution

Conceptualization, S.R.S.G. and L.S.F.; Methodology, A.V.P. and N.C.V.A.; Software,

G.M.Z.; Validation, S.R.S.G., G.M.Z., and L.S.F.; Formal analysis, A.V.P. and G.M.Z.; Research,

A.V.P., G.M.Z., and L.S.F.; Resources, C.A.H.M.; Writing-revising and editing, G.M.Z. and L.S.F.;

Visualization, C.A.H.M.; Supervision, S.R.S.G.; Project management, S.R.S.G.; Fundraising, S.R.S.G.,

A.V.P., L.S.F., and G.M.Z.

Conflict of interest

The authors declare that there are no conflicts of interest regarding the publication of this

manuscript.

Nomenclature

ABTS•+

2,2’-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)

DPPH•

2,2-diphenyl-1-picrylhydrazyl

FRAP

Ferric Reducing Ability of Plasma

Greek symbols

yij

Study variable

Overall mean

Ƭi

Effect of the ith treatment

Effect of the jth fermentation time

(Ƭi)ij(k)

Effect of the treatment-fermentation time interaction

Block effect

Effect of the ith block

eij

Experimental error

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2024 TECNOCIENCIA CHIHUAHUA.

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