Vitamin C content and amino acid identification in

carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and

MORINGA (Moringa oleifera).

Contenido de vitamina c e identificación de aminoácidos en

bebida carbonatada con kiwi (Actinidia deliciosa)

ACHOTILLO (Nephelium lappaceum) y MORINGA (Moringa

oleífera)

Jonathan Armijos-Martinez

Carolina Paz-Yépez

Abstract: The carbonated beverage industries are constantly

changing due to the growing demand for the inclusion of natural

products. Consumers are looking for beverages that are sources of

nutrients with low caloric content. In this context, an experimental

study was carried out with the main objective of determining the

vitamin C and amino acid content in a carbonated beverage made

from kiwi, achotillo and moringa. Sensory, physicochemical and

microbiological analyses were also performed on the best treatment

selected by a sensory panel of 30 untrained judges. The treatment that

received the best sensory evaluation was the one formulated with 20%

kiwifruit, 70% achotillo and 0.1% moringa. This treatment presented

a vitamin C content of 212.24 mg/kg and total amino acids of 2.29

w/w. The physicochemical results obtained were a pH of 4.3, a citric

acid content of 0.27% and total solids of 6.2 °Brix. The

microbiological results showed an absence of mesophilic aerobes, less

than 2 NPM/ml of total coliforms and less than 10 up/cm3 of molds

and yeasts. These values are within the range established by INEN

1101:2008 for carbonated beverages. Analyses of the proposed

beverage show that the product has a vitamin C content that exceeds

10% of the recommended daily intake, in addition to providing amino

acids. This makes the beverage an interesting option compared to

current carbonated beverages that do not include natural ingredients.

Keywords: achotillo, amino acids, kiwifruit, moringa, vitamin C

Published

Instituto Tecnológico Superior Edwards

Deming. Quito – Ecuador

Periodicity

April - June

Dates of receipt

Received: January 09, 2023

Approved: March 12, 2023

http://centrosuragraria.com/index.php/revista

vol. 2. Num. 18. 2023.

pp. 34-53

Correspondence author

menriquez@uea.edu.ec

Creative Commons License

Creative Commons License, Attribution-

NonCommercial-ShareAlike 4.0

International.https://creativecommons.org/lice

nses/by-nc-sa/4.0/deed.es

1 Eng. Dr. Jacobo Bucaram Ortiz Faculty of Agricultural Sciences, Agrarian University of Ecuador. Guayaquil-Ecuador,

jarmijos@uagraria.edu.ec, https://orcid.org/0000-0003-0997-1893

2 Jacobo Bucaram Ortiz, PhD Research Institute, Agrarian University of Ecuador. Guayaquil-Ecuador, cpaz@uagraria.edu.ec,

https://orcid.org/0000-0001-9547-2817

July - September vol. 1. Num. 18 - 2023

Resumen: Las industrias dedicadas a la elaboración de bebidas

carbonatadas cambian constantemente debido a la creciente demanda

de inclusión de productos naturales. Los consumidores buscan

bebidas que sean fuentes de nutrientes con un bajo contenido calórico.

En este contexto, se llevó a cabo un estudio experimental con el

objetivo principal de determinar el contenido de vitamina C y

aminoácidos en una bebida carbonatada de kiwi, achotillo y moringa.

Se realizaron también análisis sensoriales, fisicoquímico y

microbiológicos al mejor tratamiento seleccionado por un panel

sensorial conformado por 30 jueces no entrenados. El tratamiento que

recibió la mejor evaluación sensorial fue el que se formuló con un

20% de kiwi, un 70% de achotillo y un 0.1% de moringa. Este

tratamiento presentó un contenido de vitamina C de 212.24 mg/kg y

aminoácidos totales de 2.29 p/p. Los resultados fisicoquímicos

obtenidos fueron un pH de 4.3, un contenido de ácido cítrico del

0.27% y sólidos totales de 6.2 °Brix. En cuanto a los resultados

microbiológicos, se encontró ausencia de aerobios mesófilos, menos

de 2 NPM/ml de coliformes totales y menos de 10 up/cm3 de mohos

y levaduras. Estos valores se mantienen dentro del rango establecido

por la norma INEN 1101:2008 para bebidas carbonatadas. Los

análisis realizados a la bebida propuesta demuestran que el producto

tiene un contenido de vitamina C que supera el 10% de la ingesta

diaria recomendada, además de proporcionar aminoácidos. Esto

convierte a la bebida en una opción interesante en comparación con

las bebidas carbonatadas actuales que no incluyen ingredientes

naturales.

Palabras clave: achotillo, aminoácidos, kiwi, moringa, vitamina C

Introduction

In Ecuador, there are around 4,000 companies dedicated to the

production of beverages, including carbonated beverages, juices, tea,

energy drinks, water and dairy products. These beverage industries are

in a constant process of innovation with the purpose of applying the

appropriate techniques that guarantee the safety of the processes and

that in turn will help contribute to the generation of nutritional

beverages (Valverde, 2018).

Among the types of carbonated beverages are energy drinks, the energy

provided by these beverages is due to the high fructose content.

According to Maersk et al. (2012), fructose plays a very important role

in weight gain and consequently in the development of obesity. In

Ecuador, the per capita consumption of carbonated beverages is 50

l/year, due to the great demand for these beverages, there has been

interest in creating research to develop products of this type, but with

the purpose of containing and providing nutritional value, generating

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

more alternatives when consuming nutrients from a refreshing source

(Gaona-Gonzeaga et al., 2019)..

Nowadays, people try to get food products, which contain a higher

nutritional contribution and can also generate health benefits, therefore,

there has been an increased interest in the study of beverages with these

characteristics, which has contributed to the development of products

made from the mixture of vegetable species and fruit (Llanes, 2015).

Nutrients are essential for the body because they enable the

development, maintenance and functioning of our body, so by

consuming carbonated beverages we are not giving any nutritional

contribution that can benefit human health (FAO, 2019). However, this

situation can be addressed with proposals for beverages in this category

that incorporate ingredients with bioactive compounds of nutritional

interest in their formulation.

The kiwi is a fruit that has a high nutritional value, it contains vitamin

C and vitamin E, with a higher contribution of nutrients compared to

other commonly consumed fruits, vitamin C has beneficial effects on

the immune system and antioxidant power, helps in the gastrointestinal

function and the respiratory system. (Carr and Frei, 1999).

Achotillo is an exotic tropical fruit that grows in tropical or subtropical

climates. Its main component is fiber, which is resistant to digestion and

absorption in the small intestine. Among the vitamins it possesses is

vitamin C, which groups dehydroascorbic, isoascorbic and L-ascorbic

acids, which make up the active biological part that has protective

effects against various diseases such as cardiovascular diseases, it also

has phenolic compounds that help reduce the risk of diseases such as

diabetes (McEvily et al., 1992)..

Moringa is one of the fast growing plant species known as the "tree of

life" and this is due to its great nutritional contribution, moringa leaves

have a high protein content with all the essential amino acids, also, it

has a variety of vitamins such as A, B, C and E and a large number of

nutrients and micronutrients such as calcium, zinc, magnesium among

others... (Rubio-Sanz, 2020). (Rubio-Sanz, 2020)Amino acids are

essential for our organism, since they are the compounds in charge of

combining and forming the proteins that provide energy to our body.

(Solutions., 2021). Likewise, because the body does not produce

vitamin C, it needs to obtain it from the diet (Carr and Maggini, 2017).

Based on the above, the importance of generating alternatives to the

consumer with regard to innovative products that provide nutrients is of

July - September vol. 1. Num. 18 - 2023

greater interest every day. As well as in the proposal to generate a

positive change in the choice of beverages that denote a benefit.

Rodriguez (2020) determined the presence of antioxidants in a beverage

made with soy milk (Glycine max) and kiwi (Actinida deliciosa),

flavored with passion fruit. From this study it is highlighted that the best

sensorially qualified treatment of the soy beverage was the one

elaborated with 40 % soy, while, of the kiwi concentrations studied, the

treatment with the highest sensorial acceptance was the one containing

40 % kiwi.

Sanchinelli (2004) conducted an investigation to determine the protein

content, essential amino acids in leaves, flowers and stems of Moringa

oleifera, which is cultivated in Guatemala. The protein content in

Moringa oleifera was found to range from 1.30 to 4.62 %, with the

highest content in the leaves. The essential amino acid content was

found to range from 98 to 1384 mg/100 g and from 90 to 270 mg/100 g

in dehydrated and fresh leaves, respectively. In flowers the range was

from 23 to 122 mg/100 g and in stems from 11 to 37 mg/100 g. The

limiting amino acid in leaves was lysine; in flowers the limiting amino

acids were valine and methionine; and in stems the limiting amino acid

was valine.

Cedaro et al. (2015) presented a paper on the analysis of vitamin C in

soft drinks, the results of a survey carried out on 52 products that are

part of the usual offer of these beverages Obtained ascorbic acid levels

in a range from 0 to 82.36±4.84 mg/200mL, corresponding the highest

values to orange flavored products. This study determined the presence

of vitamin C and the amino acid profile in a carbonated beverage with

kiwi (Actinidia deliciosa), achotillo (Nephelium lappaceum) and

moringa (Moringa oleifera) that will provide a beverage with

nutritional appeal. The antioxidant properties of kiwi and achotillo,

together with the nutritional benefits such as the amino acids provided

by moringa, will be used to generate a new carbonated beverage that

not only provides the sensation of freshness, but is also potentially

beneficial to health.

Materials and methods

The present work proposed the study of the vitamin C content and

amino acid profile in a carbonated beverage composed of three

ingredients: kiwi, achotillo and moringa. The details of the proposed

formulation are presented in Table 1.

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

Table 1. Percentages of ingredients used for the different treatments in

the formulation of the carbonated beverage.

Description of the percentages of ingredients for the different

treatments in the formulation of the carbonated beverage.

Armijos, 2022

From the results obtained, the beverage with the best score was selected.

The formulated beverage with the best sensory score had its vitamin C

content, amino acid profile, physical-chemical and microbiological

analysis determined.

Material resources

The raw materials (kiwi, achotillo, moringa, monkfruit, lemon, sodium

benzoate, carbon dioxide) used were obtained from local businesses.

Methods and techniques

Figure 1 shows the flow diagrams of the processes used to obtain the

beverage under study.

Sensory evaluation method

Ingredients

Actinidia

deliciosa (kiwi)

25%

20%

15%

Nephelium

lappaceum

(Achotillo)

45%

50%

55%

Moringa oleifera

0.1

0.1%

Monk fruit

0.25

0.25%

Water

29.04%

Sodium benzoate

Citric acid

0.5%

0.05%

0.06%

0.5%

0.05%

0.06%

0.5%

0.05%

0.06%

July - September vol. 1. Num. 18 - 2023

For the sensory evaluation of the organoleptic characteristics (color,

odor and flavor), a panel composed of 30 untrained persons between 18

and 30 years of age was selected; each member of the panel received 3

containers, each containing a treatment to be evaluated. Subsequently,

they received an evaluation form where they recorded the data using a

5-point hedonic scale, where 5 is the highest score and 1 is the lowest

score.

The score of 5 had a category of "I like it very much", the score of 4

had a category of "I like it", the score of 3 had a category of "I neither

like nor dislike it", the score of 2 had a category of "I dislike it" and the

score of 1 had a category of "I dislike it very much". To evaluate the

vitamin C content, amino acid profile, physicochemical and

microbiological parameters of the best evaluated treatment, analyses

were performed in an accredited laboratory.

Physicochemical parameters

pH determination

For pH titration, the procedure detailed in INEN 1087 was followed.

Determination of carbon dioxide gas

For the determination of carbon dioxide gas, the procedure detailed in

INEN 1082 was followed.

Determination of Brix degrees

For the determination of Brix degrees, the procedure detailed in INEN

1083 was followed.

Acidity determination

For the determination of acidity, the procedure detailed in NTE INEN

1091 was followed.

Determination of vitamin C by HPLC

The sample chosen with the best score by the sensory panel had its

vitamin C content determined by HPLC.

Liquid chromatography (HPLC) is a technique used to separate the

components of a mixture. It consists of a non-polar stationary phase

(column) and a mobile phase. The stationary phase is silica that has

been treated with RMe

SiCl. The mobile phase acts as a sample carrier.

The sample in solution is injected into the mobile phase. The

components of the solution migrate according to the non-covalent

interactions of the compounds with the column. These chemical

interactions determine the separation of the contents of the sample. The

use of the different detectors will depend on the nature of the

compounds to be determined (Miranda and Martín, 2013).

C18 column and an RP-18 precolumn were used for the stationary

phase, and the mobile phase, sulfuric acid was used to reach pH 2.2.

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

The flow rate was 0.4 mL/min, and the UVVIS detector at 225 nm.

(Fang, 2017).

Determination of amino acid profile by the Burbach Institute

Prederivatized (HPLC-FLD) method.

The sample chosen with the best score by the sensory panel had its

amino acid profile determined by the Burbach Institute Prederivatized

(HPLC-FLD) method (Castillo-Portela et al., 2011). (Castillo-Portela et

al., 2011)..

Microbiological analysis for mesophiles by BAM-FDA method

CAP#3

This method is based on the certainty that a vital microorganism present

in a food sample, when inoculated in a solid nutrient medium, will

reproduce forming a visible individual colony. To make colony

counting possible, decimal dilutions of the initial sample suspension are

made and the nutrient culture medium is inoculated. The inoculum is

incubated at 30°C for 72 hours and then the number of colonies formed

is counted. The count is used to calculate the amount of microorganisms

per gram or per cubic centimeter of food (INEN, 2006). (INEN, 2006).

The sample chosen with the best score by the sensory panel underwent

the respective microbiological analysis by the most probable number

method for the detection of mesophiles.

Multiple fermentation tube method for coliform analysis

The method is based on the detection of the most probable number

(MPN) by the tube dilution technique using the selective liquid medium

bile-lactose brilliant green broth or similar for the presumptive assay

and the tubes that present gas are plated on Eosin methylene blue agar

(E M B). The incubation temperature for the presumptive and

confirmatory assay is 30 +-1°C for refrigerated products and 35+-1°C

for products kept at room temperature. (INEN, 1990).

Analysis of molds and yeasts by the plate count method by deep

sowing

This method is based on the cultivation between 22ºC and 25ºC of the

propagating units of molds and yeasts, using the plate count technique

by deep sowing and a medium containing yeast extract, glucose and

mineral salts. (INEN, 2013).

Sensory evaluation method

A sensory panel of 30 untrained judges was selected for the analysis of

odor, color and flavor of the samples of each of the treatments under

July - September vol. 1. Num. 18 - 2023

study. A 5-point hedonic scale was used to measure the sensory

characteristics.

Statistical analysis

The information obtained at the end of the carbonated beverage

treatments for each of the sensory variables, using the 5-point scale

planned to be used, was statistically evaluated by analysis of variance

(ANOVA). In the case of obtaining significant differences, the Tukey

test was used as a test for comparison of means. These two statistical

tools were applied at 5% significance.

The analysis of variance model is detailed in Table 2.

Table 2. Analysis of variance for sensory variables

Sources of variation

Degrees of freedom

Total (n-1)

Treatments (t-1)

Repetitions (judges)(R-1)

Experimental error (t-1) (R-1)

Presentation of analysis of variance for sensory variables, Armijos,

2021

3. Result

Results of formulation selection of carbonated beverage

formulation made with kiwi achotillo and moringa from a sensory

panel.

Sensory tests were conducted to evaluate a carbonated kiwi, achotillo

and moringa beverage. Three treatments were established with different

proportions of kiwifruit and achotillo, while moringa, monkfruit, citric

acid and sodium benzoate were held constant in the three formulations.

The preparation of the beverage followed the process previously

detailed in Figure 1. 20 kg of kiwifruit and achotillo in good condition

and of acceptable quality were selected, based on their physiological

maturity index and sugar content (12ºBrix for kiwifruit and 17ºBrix for

achotillo). The fruits were washed with water and disinfected with 0.5%

lactic acid for 3 minutes. They were then peeled and the pulp was

weighed according to the proportions required for each formulation.

The pulps were liquefied to obtain the juice, which was then filtered to

separate solid residues.

Subsequently, the pulps were mixed and citric acid was added as a

stabilizer and sodium benzoate as a preservative. Pasteurization was

carried out at 70ºC for 30 minutes, followed by thermal shock in water

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

and ice to rapidly reduce the temperature to 20ºC and ensure the

elimination of microorganisms. Water, non-caloric sweetener

(monkfruit) and moringa powder were added. The juice was cooled to

4ºC for one day and then carbonated by mixing with carbon dioxide.

Finally, it was packaged in amber glass bottles and stored at 4ºC.

Analysis of variance and tukey's test to the treatments evaluated by

the panelists.

The results obtained from the sensory tests of the treatments performed

on the panelists with their replicates were recorded in a Microsoft Excel

spreadsheet, and then exported to Infostat (2020), followed by an

analysis of variance using the Tukey test at 5% probability, which made

it possible to determine the significant difference between treatments

by checking the significance level (p-value).

As can be seen in Table 4, treatment 2 did not present a significant

difference with treatment 1 for the attributes of color and flavor, so

treatment 2 was chosen for having the highest mean, which was 3.73

for the attribute of color, while the mean for this same treatment in the

attribute of flavor was 3.60 and for the attribute of odor, treatment 2 had

a mean of 3.50, presenting a significant difference between treatments

1 and 3, which did not present a significant difference between them

with respect to the aforementioned attribute.

Table 3 shows the results of the sensory evaluation of the treatments.

Table 3. Sensory evaluation of the kiwi, achotillo and moringa

carbonated beverage treatments.

Means of sensory attributes in the proposed treatments of the

carbonated beverage.

Means with a letter in common are not significantly different (p>0.05).

Treatments Color Odor Flavor

Treatment 1 3.57

2.93

3.39

Treatment 2 3.73

3.50

3.60

Treatment 3 3.06

2.86

CV 27.91 37.89 35.92

July - September vol. 1. Num. 18 - 2023

Determination of the Vitamin C content and amino acid profile of

the carbonated beverage made with kiwi, achotillo and moringa

selected as the best scored from the sensory panel.

According to the analysis of the vitamin C content of the kiwi, achotillo

and moringa carbonated beverage corresponding to treatment 2,

selected as the one with the highest sensory acceptance, it registered a

vitamin C content of 212.39 mg/kg (21.24 mg/100g), highlighting that

the raw materials used for its preparation are a good source of this

vitamin.

Amino acid profile results of the best scoring treatment of the kiwi,

achotillo and moringa carbonated beverage.

Table 4 shows the values corresponding to the amino acid profile of

treatment 2 of the carbonated beverage selected as the best scored by

the sensory panel.

Table 4. Amino acid profile of carbonated beverages

Results of the amino acid profile of the carbonated beverage.

Evaluation of the physicochemical (acidity, ºbrix, pH, CO Volume

)

and microbiological (total& coliforms,& mesophilic& molds& and&

yeasts)& parameters of the kiwi (Actinidia deliciosa) achotillo

(Nephelium lappaceum) and moringa (Moringa oleifera) carbonated

Amino acids gAA/100g

Aspartic Acid 0.18

Glutamic Acid 0.35

Serine 0.12

Histidine 0.00

Threonine 0.12

Glycine 0.16

Arginine 0.12

Alanine 0.00

Tyrosine 0.72

Valine 0.09

Methionia 0.04

Phenyl alanine 0.12

Isoleucine 0.09

Leucine 0.13

Lysine 0.05

Total amino acids 2.29 w/w

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

beverage according to the NTE INEN 1101:2008 Standard for

carbonated beverages for the treatment with the highest score in the

sensory panel.

Physicochemical results of the kiwi, achotillo and moringa

carbonated beverage.

The results of the physicochemical parameters for treatment 2 indicate

that they are within the ranges established by NTE INEN 1101:2008 for

carbonated beverages.

Table 5 presents the results obtained from the physicochemical

analysis:

Table 5. Physicochemical parameters of the treatment with the highest

acceptance by the sensory panel.

Physicochemical analysis of the best-scoring carbonated beverage

treatment

Armijos, 2022

Results of the microbiological analysis of the best-scoring

treatment of the kiwi, achotillo and moringa carbonated

beverage.

The results of the microbiological analyses performed on treatment 2

of the kiwi, achotillo and moringa carbonated beverage indicate that

they are within the microbiological parameters established by NTE

INEN 1101:2008 for carbonated beverages. Table 6 shows the results

of the microbiological quality analysis of the carbonated beverage:

Table 6. Results of microbiological analysis of the treatment with the

highest acceptance by the sensory panel.

Parameters

Results

Acidity

0.27%

Brix

Volume CO2

6.2

4.3

2.6

July - September vol. 1. Num. 18 - 2023

Microbiological analysis results

Armijos, 2022

For the present study, the vitamin C content and amino acid profile of

a carbonated beverage with kiwi, achotillo and moringa were

determined, for which three different treatments were formulated with

different percentages as indicated in previous sections (T1: 25% kiwi,

45% achotillo, 0.1% moringa, 0.25% monkfruit and 0.25% sodium

benzoate).1% moringa, 0.25% monkfruit and 0.25% sodium benzoate)

T2: 20% kiwifruit, 50% achotillo, 0.1% moringa, 0.25% monkfruit and

0.25% sodium benzoate) T3: 15% kiwifruit, 55% achotillo, 0.1%

moringa, 0.25% monkfruit and 0.25% sodium benzoate). According to

the results obtained from the sensory panel, the most acceptable

treatment in terms of color, odor and flavor attributes was treatment 2,

which was analyzed for vitamin C content, amino acid profile,

physicochemical analysis (acidity, pH, °brix and volume of CO )

and

microbiological analysis (total coliforms, mesophiles, molds and

yeasts). In a study conducted by Gonzaga (2017), where he conducted

sensory analysis of a carbonated beverage made from naranjilla pulp,

the formulation of the beverage made up of 25% naranjilla pulp and

10% sweetener obtained greater acceptance; this formulation had a

greater amount of pulp extract, which allowed it to maintain the

characteristic flavor, color and smell of the fruit; however, the panelists

recommended reducing the intensity of the sweet flavor. Based on the

above, it can be indicated that in the carbonated beverage proposed with

kiwi, achotillo and moringa, a value of 2% was considered for the

sweetener based on the juice of the fruits that were added, so that it

would not be affected in terms of the sweet flavor and thus could

highlight a characteristic flavor of the fruits used in the beverage.

Rodriguez (2020) evaluated the organoleptic characteristics of a

beverage made from soy milk and kiwi, flavored with passion fruit. The

Requirements

Unit Results Test method

Total coliforms

Mesophilic

Molds and yeasts

NMP/ml <3 PEE.LASA.MB.01b;BAM

CAP.

4,ED.2017

CFU/g 3.0x10

BAM-FDA CAP.#3

2001

(Plate count)

CFU/g <10 INEN 1529-10 1998

(Plate count)

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

best qualified treatment of this beverage was treatment 4 made with

40% soy and 40% kiwi, while the remaining treatments showed no

significant difference between them, with measurements well below the

winning treatment. Similar results were shown in the sensory evaluation

of the kiwi, achotillo and moringa carbonated beverage, where it was

shown that the treatment with the highest sensory mean was treatment

2, but in this case it did not show a significant difference with treatment

1, for which the highest mean corresponding to T2 was chosen.

The analysis to determine the vitamin C content of treatment 2, chosen

by the sensory panel for being the highest scoring, resulted in a value

of 212.39 mg/kg (21.24 mg/100g), a value higher than the vitamin C

content obtained by Maldonado and Moncayo (2012), in their

carbonated passion fruit drink where they recorded a vitamin C value

of 10.02 mg/100g of vitamin C, this result is surpassed by the proposed

carbonated drink of kiwi, achotillo and moringa because the matrices

used have a high vitamin C content, highlighting for example 70

mg/100 g is the vitamin C content of achotillo, 93 mg/100 g is that of

kiwi and 70 mg/100 g in moringa, all these matrices exceed the fruit

used by Maldonado and Moncayo (2012), which contains 30 mg/100 g

of this vitamin. On the other hand, in a study conducted by Cantillo

(2020), where he elaborated nectar based on achotillo and apple

enriched with alfalfa as a nutritional contribution, he evaluated the

vitamin C content by the method of volumetric titration to several

treatments in which the control treatment consisting of 50% achotillo

and 50% apple obtained a higher vitamin C content which was

26.25mg/100 ml, followed by the treatment consisting of alfalfa 2.5%,

achotillo 40% and apple 57.5% with a vitamin C content of 24.13

mg/100 ml and the lowest value was the treatment made up of 5%

alfalfa, 50% achotillo and 45% apple which was 18.25/100 ml,

resembling these vitamin C contents with that of the present project

under study which includes one of the fruits used (achotillo) by the

authors mentioned above, this being a carbonated beverage. It is

important to highlight that pasteurization is a treatment that affects the

loss of vitamin C as confirmed by Villareal, Mejía, Osorio, and Ceron

(2013), who analyzed the effect of pasteurization on sensory

characteristics and vitamin C content in fruit juice, which were tree

tomato juice, costeño mango juice, valencia orange and castilla

blackberry, initially these juices had a vitamin C content of 6.4mg/100g,

7.2mg/100g, 55mg/100g and 2.4mg/100g, after being subjected to

July - September vol. 1. Num. 18 - 2023

different temperatures and pasteurization times for each juice,

corresponding as follows, blackberry juice T=80°C and t=120 s, mango

juice T=85°C and t=120 s, tree tomato juice T=80°C and t=90 s and

orange juice T=75°C and t=90 s, evidenced a loss of vitamin C in each

fruit juice, for tree tomato juice the loss was 87.72%, mango juice

89.82%, orange juice 99.43% and blackberry juice 84.67%. On the

other hand, Rodriguez (2007), elaborated and determined the stability

of an isotonic drink based on Kiwi and Guava added with conjugated

linoleic acid (CLA), where the fruits with peel were subjected to a steam

scalding at 80°C for 30 minutes, the analysis of vitamin C was carried

out by the spectrophotometric method, obtaining as a result a vitamin C

content of 41.This value is almost double that found in the carbonated

kiwi achotillo and moringa drink with a vitamin C content of 21.24

mg/100 g. It is important to note that guava has 3 times more vitamin C

than kiwifruit and almost 4 times more than achotillo, and the heat

treatment applied to the isotonic beverage influences the vitamin C

value to be higher than that found in the carbonated kiwifruit achotillo

and moringa beverage, which was pasteurized at 75°C for 25 minutes.

The results for the amino acid profile of the kiwi achotillo and moringa

carbonated beverage of treatment 2 chosen as the most acceptable by

the sensory panel, recorded the following results; Aspartic Acid 0.18

g/100 g (1.8mg/g sample), Glutamic Acid 0.35 g/100 g (3.5 mg/g

sample), Serine 0.12 g/100g (1.2 mg/g sample), Threonine 0.12 g/100g

(1.2 mg/g sample), Glycine 0.16 g/100 g (1.6 mg/g sample), Arginine

0.12 g/100 g (1.2 mg/g sample), Tyrosine 0.75g/100 g (7.5 mg/g

sample), Valine 0.09 g/100 g (0.9 mg/g sample), Methionine 0.04 g/100

g (0.4 mg/g sample), Phenyl alanine 0.12 g/100 g(1.2 mg/g sample),

Isoleucine 0.09 g/100 g (0.9 mg/g sample), Leucine 0.13 g/100 g (1.3

mg/g sample), Lysine 0.05 g/100 g (0.5 mg/g sample), with total

essential amino acids (lysine, leucine, threonine, isoleucine,

methionine, valine, phenylalanine) of 0.64 g/100 g except for Histidine

and Alanine, results that are well below that evidenced by Santos

(2019), who performed a characterization of a Tea by using moringa

leaves which determined the amino acid profile in moringa leaves

dehydrated at different temperatures 35, 45 and 55°C characterizing the

amino acid profile by the HPLC method where the highest content of

essential amino acids was identified in the treatment at 55°C obtaining

as a result 16.It is important to highlight that the author mentions that

the heat applied to the fresh moringa leaves to dehydrate them

influenced the concentration of amino acids, because there was a greater

loss of water in the leaves when the temperature of 55°C was applied,

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

so that at this temperature the concentration of amino acids was not high

enough.

In another study conducted by Vendrell (2021), on the protein and

antioxidant enrichment of cookies with moringa dried leaves powder,

where he added moringa powder to wheat flour in different proportions

called G1 and G10, G1 consisted of 1% moringa powder and 53%

wheat flour while G10 consisted of 10% moringa powder and 44%

wheat flour in this study determined the amino acid profile of the cookie

made with these formulations using the gas chromatography method,

the highest results of amino acids were given in the formulation G10

which were; for Isoleucine 0.02 g/100 g sample, Aspartic Acid 0.33

g/100 g sample, Glutamic Acid 0.45 g/100 g sample, Serine 0.18 g/100g

sample, Threonine 0.1 g/100g sample, Proline 0.015g/100 g sample,

Asparagine 0.03 g/100 g sample, Hydroxyproline 0.01 mg/100 g

sample, Tyrosine 0.02 g/100 g sample, Methionine 0.09 g/100 g sample,

Phenylalanine 0.02 g/100 g sample, α-aminoadipic acid 0.6 g/100 g

sample, as can be evidenced the results only contain 3 of the essential

amino acids ( Isoleucine 0.02 g/100 g sample , Methionine 0.09 g/100

g sample and Phenylalanine 0.02 g/100 g of sample), these quantities

are lower than the essential amino acids found in the carbonated kiwi,

achotillo and moringa beverage, of which Isoleucine with 0.09 g/100 g

and Phenylalanine with 0.12 g/100 g are found in greater proportion.

This loss of essential amino acids in the case of the cookie is due to the

high baking temperature (180 ºC) applied for the elaboration of the

cookie, as stated by the author Vendrell (2021).

The results obtained for the physicochemical parameters of the kiwi

achotillo and moringa beverage of treatment T2 selected as the most

acceptable by the sensory panel were 6.2 °brix, 4.3 pH, titratable acidity

0.27% and CO2 volume of 2.6 /L, which are within the range

established by the INEN (2008) standard for low-calorie carbonated

beverages, however, in the physicochemical results obtained by

Gonzaga (2017), where he elaborated a carbonated beverage with

naranjilla pulp presented 10°brix, pH 3.5, titratable acidity 0.55% and

CO2 volume of 2.5 /L, this beverage reflects higher values in terms of

acidity and pH, this is due to the fact that naranjilla has an acidity of

2.56%. Gonzaga (2017)being more acidic than kiwi with 1.18% Godoy

and Dome (2013), and achotillo with 0.30% acidity Avendaño-Arrazate

et al., (2017), fruits that were occupied for the elaboration of the

carbonated beverage of the present research.

July - September vol. 1. Num. 18 - 2023

Finally, microbiological parameters were evaluated for treatment 2,

based on the INEN (2008) standard for carbonated beverages, showing

results for total coliforms < 3 (Absence), Mesophiles < 1/d (Absence),

molds and yeasts < 10, showing that all these values are in accordance

with the specifications established by the INEN 1101 standard:2008 for

carbonated beverages, it should be mentioned the importance of

performing microbiological analyses since they determine the safety of

the process and can affect the shelf life and sensory parameters of the

beverage.

4. Conclusions

According to the sensory analysis carried out on the carbonated

beverage made with kiwi (15%), achotillo (25%) and moringa (0.1%),

treatment 2 was chosen as it presented the greatest acceptance by the

sensory panel with respect to the attributes of color, odor and flavor,

evaluated with the help of the infostad program, where it was

determined through the analysis of variance that treatment 2, although

it did not present a significant difference with treatment 1, was chosen

as it showed the highest mean among the three treatments carried out.

From the results obtained by the laboratory with respect to vitamin C

and the amino acid profile of treatment 2 of the carbonated beverage,

212.39 mg/kg (21.24 mg/100 g) of vitamin C content was obtained, this

amount would represent approximately 20% of the recommended daily

value of vitamin C indicated as a voluntary declaration for nutrients in

INEN 1334-2 Labeling of Food Products for Human Consumption Part

2: Nutritional Labeling. Requirements (NTE INEN 1334-2, 2011).

In relation to the amino acid profile, it could be evidenced that the

beverage presents a total of 2.29 AAg/100 gram of sample, being able

to highlight this presence of nutrients when compared with other

carbonated beverages in the market. It should be emphasized that for

this research only 0.1% of moringa was used, which is a good source of

amino acids, but for formulation purposes the amount added was low

due to its intense flavor and aroma, which is not accustomed to the

palate of consumers, which altered the sensory quality of the proposed

beverage.

The results of the physicochemical parameters for the kiwi, achotillo

and moringa carbonated beverage of treatment 2 were 6.2 ° Brix, 4.3

pH, titratable acidity 0.27% and CO2 volume of 2.6 /L. These results

show that they are within the ranges established by the INEN 2008

standard for carbonated beverages. With respect to the microbiological

results for mesophilic aerobes, total coliforms, molds and yeasts, the

Vitamin C content and amino acid identification in carbonated beverage with kiwifruit (Actinidia

deliciosa), ACHOTILLO (Nephelium lappaceum) and MORINGA (Moringa oleifera)

carbonated beverage complies with the parameters required by the NTE

INEN 2008 standard.

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