Antioxidant Analysis in Drinks of Guava, Lemon, Melon, and Beet

Juice Fermented Using Lactobacillus Plantarum

Afrinia Eka Sari

, Reza Anindita

and Wening Cahyani

Department of Nutrition STIKes Mitra keluarga, Bekasi, Indonesia

Department of Pharmacy STIKes MItra Keluarga, Bekasi, Indonesia

Keywords: Antioxidant, Activity, Probiotic, Jalembi.

Abstract: Antioxidants are active substances that can counteract free radicals, where free radicals can trigger infectious

and non-communicable diseases. Antioxidants are found in many vegetables and fruits. Jalembi (Jambu,

Lemon, Melon, Bit) drink is an alternative source of antioxidants that can counteract free radicals. Guavas,

lemons, melons, beets are fruits that contain vitamin C, which is a substance that acts as an antioxidant. The

purpose of this research to determine the antioxidant contain of fermented Jalembi drink using Lactobacillus

Plantarum. The reason for using lactobacillus plantarum in this research is because lactic acid bacteria are

commonly used in the fermentation process and also have the advantage of being resistant to acidic pH and

several studies have stated that these bacteria can survive in the digestive tract. The research design is an

experimental study with a completely randomized design. The three formulas differ primarily in sugar content,

with formula 1 containing 10% sugar, formula 2 containing 20%, and formula 3 containing 30%. The results

of the analysis of antioxidant activity in the three formulas were very weak or there was no antioxidant

activity. In terms of toxicity analysis, jalembi probiotic drink is safe for consumption and the average panelist

acceptance results of the three formulas are quite favorable. Suggestions for further research should test the

antioxidant contain without bacteria fermentation process.

1 INTRODUCTION

Basic Riset Kesehatan Dasar (Riskesdas) 2018

shows 95.5% of Indonesian people consume less

vegetables and fruit. The average consumption of the

Indonesian people is 81.14 grams/capita/day (BPS,

2021). However, this amount is only 54.09% of the

minimum nutritional adequacy rate. Fruits and

vegetables not only contain vitamins, minerals and

fiber but also contain antioxidants.

Antioxidants have a function to ward off free

radicals. Free radicals are defined as molecules

containing one or more electrons which are highly

reactive and can cause cell damage or death (aging).

Free radicals can also trigger non-communicable

diseases such as hypertension, stroke, obesity and

diabetes mellitus. As a result of continuous exposure

to free radicals and low intake of vitamin C and

vitamin E can lead to depletion of natural antioxidant

production in the body, causing various diseases.

Physical activity with low and moderate intensity is

needed to increase the antioxidant system because it

maximizes free radical removal (Sinaga, 2018).

Antioxidants are compounds that are able to

counteract or reduce the negative effects of oxidants

in the body, working by donating an electron to

compounds that are oxidants so that the activity of

these oxidant. This research aimed to determine the

antioxidant in Jalembi drinks ousing red guava,

lemon, melon, and beet juice and L. plantarum

starter.

2 MATERIALS AND METHOD

The material ingredients used in the development of

Jalembi drinks are guava fruit, lemon, melon,

beetroot extract, water, guava flavor, granulated

sugar, and Lactobacillus plantarum compounds can

be inhibited (Wulan, 2019). The fruit used are fruit

that have sufficient level of ripeness.

Factors that can affect antioxidant activity in a

food include pH, temperature, light, and adding sugar.

Factors that can affect antioxidant activity in a food

include pH, temperature, light, and adding sugar.

Based on research conducted by Fidyasari et al.

Sari, A. E., Anindita, R. and Cahyani, W.

Antioxidant Analysis in Drinks of Guava, Lemon, Melon, and Beet Juice Fermented Using Lactobacillus Plantarum.

DOI: 10.5220/0012919600004564

Paper published under CC license (CC BY-NC-ND 4.0)

In Proceedings of the 5th International Conference on Social Determinants of Health (ICSDH 2023), pages 209-212

ISBN: 978-989-758-727-6; ISSN: 2975-8297

209

(2019), lactic acid bacteria in probiotic drinks can

provide antioxidant activity. Fermented drinks can

also increase flavonoid compounds as a source of

antioxidants.

One of the bacteria used in this study

is Lactobacillus plantarum. Lactobacillus

plantarum is a lactic acid bacterium with the

potential as a biopreservative because it can inhibit

the growth of pathogenic and destructive

bacteria. Lactobacillus plantarum can inhibit the

growth of pathogenic bacteria with the most

significant inhibition compared to other lactic acid

bacteria (Hariati, 2019).

The tools used in the development of probiotic

drinks are knives, cutting boards, measuring cups,

blenders, filters, pans, stoves, spoons, digital scales,

basins, glass bottles, pH meters, autoclaves for bottle

sterilization, and incubator for incubation. The type

of research design used in this research is an

experimental method using Completely Randomized

Design (CRD).

The analysis included hedonic tests and

antioxidant tests. The research was conducted from

January to March 2023 in STIKes Mitra

Keluarga,using thirty people panelist.

Sample Preparation. First, wash all the fruit,

then peel it. Then, weigh all the ingredients according

to the formula, crush it with a blender, strain it until

there is no pulp, and heat it at 70 degrees Celsius for

10 minutes. The sample that has been heated is cooled

and transferred into a sterilized bottle. Then, add 1 ml

of Lactobacillus Plantarum bacteria starter. There

were three formulas used in this study with a

comparison of guava: lemon, melon, beets, flavors,

bacterial starter (40:10:27:3:4:1). The variations lie in

the amount of added sugar: formula 1 includes 10

grams, formula 2 include 20 grams, and formula 3

includes 30 grams. Before the incubation process is

carried out, sterilization of equipment and containers

must be ensured, and sterilization is carried out at a

temperature of 121 degrees Celsius.

The heated sample is cooled and transferred to

a sterilized glass bottle, followed by the addition of

1ml of Lactobacillus Plantarum bacteria starter.

Incubate at 37 degrees Celcius for 24 hours..

Organoleptic test results data were analyzed

using a non-parametric statistical method, namely the

Friedman test. Friedman's test is used to analyze the

comparison of three or more averages in dependent

samples (paired) where the assumption of normality

is not met. In this study, the Friedman test was used

to determine the difference between the control group

and the experimental group. In this study, the

Wilcoxon test was also used to compare the result of

organoleptic and hedonic test without having to see

whether the data was normally distributed.

3 RESULT AND DISCUSSION

Based on the result of measuring the acidity level, the

following result were obtained.

Table 1: pH level of Jalembi drink.

Treatment pH Criteria

Formula 1 3,43 Acid

Formula 2 3.39. Acid

Formula 3 3.32 Acid

Source: Primary date,2023

The three formulas have a fairly high level of

acidity which is caused by lactic acid produced from

the fermentation process.

The results of the hedonic test for the three

formulas did not show significant differences:

however, whwn taste preferences were examined.

Formula 3 emerged as the most preferred. The

following below is a table of hedonic test results.

Table 2: Hedonic test.

Indicator

Criteri

Treatment Colour Odor Taste Viscosity

Formula 1 3.85 3.65 3.30 3.50 Like

Formula 2 3.40 3.53 3.93 3.45 Like

Formula 3 3.95 3.53 4.03 3.58 Like

Source: Primary date,2023

The hedonic test determines panelists'

preferences based on indicators of color, aroma,

taste, and viscosity (viscosity). The hedonic test scale

for probiotic beverage products starts from numbers

1-5. Description of the scale on the level of

preference or hedonic.

Data is said to be abnormal if it has p <0.05. If

p<0.05, there is a significant difference, and from

that, the data analysis is continued with the Wilcoxon

test. If the Wilcoxon test is p <0.05, each formula has

significant differences. Test, namely value (1) means

dislike, value (2) means dislike, value (3) means

quite like, value (4) means like, and value (5) means

really like. Based on the color criteria, the color

contained in the jalembi probiotic drink comes from

the betalain compound in beetroot extract, which

produces an attractive red or purple color, making it

a natural dye. This is by the opinion of Pramesti

(2019), which states that determining the quality of

food ingredients is generally influenced by several

ICSDH 2023 - The International Conference on Social Determinants of Health

210

factors, including taste, texture, and nutritional value

Based on the taste criteria, the probiotic drink tastes

sour and sweet.

The sour taste is obtained from the fermentation

of LAB Lactobacillus plantarum, while the sweet

taste is obtained from adding sugar in each sample.

The probiotic drink sample that the panelists liked

the most in formula 3 was due to the addition of 30%

sugar, resulting in a sweet taste that was more

attractive to consumers, while the product that was

less liked by the panelists was in the control

treatment, namely products without the addition of

sugar so that the resulting taste was sour. The results

of the panelist's preference analysis on the viscosity

parameter (viscosity) in formula 1 have a value of

3.50; formula 2 has a value of 3.45; formula 3 has a

value of 3.58 and the control treatment has a value of

3.15, where the four formulas fall into the like

category. The aroma of this probiotic drink is

obtained from the aroma of guava fruit, which is

more dominant because it adds guava flavor so that

the resulting aroma is sweet and fragrant. Aroma is a

determinant of the quality of a food or beverage

product. The sense of smell can feel a distinctive

aroma depending on the ingredients added to the

drink (Feriady, 2018)

Antioxidant activity test using the

spectrophotometric DPPH method. The results of the

analysis of the antioxidant activity test in each

sample can be seen in the table below.

Table 3: Antioxidant.

Sample Antioxidant activity Criteria

Formula 1 3586.23 ppm

No antioxidant

activity

Formula 2 2178.48 ppm

No antioxidant

activity

Formula 3 3785.20 ppm

No antioxidant

activity

The results of the analysis on the antioxidant

activity of the probiotic drink (guava, lemon, melon,

beet) showed that the highest level of antioxidant

activity was in the treatment of formula 2 (20 grams

of added sugar) with an antioxidant activity level of

2178.48 ppm. The lowest result was found in formula

3 (30 grams of added sugar) with an antioxidant

activity level of 3785.29 ppm, which, according to

theory, means that the smaller the IC50 value, the

higher the antioxidant activity. (Wulansari, 2018).

The principle of the antioxidant activity test

method is to measure the scavenging of DPPH

radicals by a compound that has antioxidant activity

using UV-Vis spectrophotometry so that the value of

free radical scavenging activity will be known, which

is expressed by the IC50 value. IC50, namely the

amount of inhibitory concentration of the test solution

on its ability to reduce free radical activity by 50%

(Wulansari, 2018).

The study's results on jalembi probiotic drinks

found that the antioxidants in the three formulas

showed weak activity or did not have antioxidant

activity. As for what inhibits the activity of

antioxidants in the product, include adding sugar.

This by glycosylation reactions, where sugar interacts

with antioxidant compounds. This reaction produces

glycosylation products that reduce the availability of

hydrogen atoms (H) and methyl groups in antioxidant

compounds. H atoms and methyl groups play an

important role in antioxidant activity, especially as

hydrogen donors to unstable free radicals. This loss

of H atoms reduces the ability of antioxidant

compounds to act as hydrogen donors against free

radicals.

As a consequence, antioxidant activity decreases

due to the lack of ability of the compounds to capture

and neutralize free radicals. (Pelealu, 2019). Another

factor that affects antioxidant activity is the length of

testing after incubation. After 2-4 days of incubation,

the antioxidant activity increases, then on the fifth

day, there will be a drastic decrease (Putro H, 2020).

Apart from the influence of the glycosylation

reaction, the decrease in levels of antioxidant activity

can be caused by the heating process. The ingredients

used in the manufacture of this probiotic drink

product are ingredients that contain vitamin C. It is

known that vitamin C is a substance that can act as an

antioxidant. Vitamin C in the probiotic drink tends to

be more susceptible to damage at high

temperatures.Vitamin C can experience degradation

and lose its antioxidant activity when exposed to

temperatures of 70-90 ºC or during a long heating

process

Based on data analysis using the Friedman test,

the results for the aroma and viscosity indicators

showed no significant difference because the p-value

was > 0.05. The aroma indicator has a p-value of

0.362 and the viscosity indicator has a p-value of

0.378. As for the color and taste indicators, there is a

significant difference because they have a p-value

<0.05. Where the color and taste indicators have a p-

value of 0.0001, this shows that adding sugar affects

the color and taste of the probiotic drink.

Antioxidant Analysis in Drinks of Guava, Lemon, Melon, and Beet Juice Fermented Using Lactobacillus Plantarum

211

4 CONCLUSION

Organoleptically, based on taste, color, aroma, and

viscosity, the probiotic drink is quite preferred and

has the potential as a health drink as a source of

probiotics. Further research is needed to determine

the factors that cause a decrease in antioxidant

activity in probiotic drinks.

ACKNOWLEDGMENTS

Our gratitude is sent to STIKes Mitra Keluarga, who

funded this research, and thank for this research for

the work team.

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