Sugar Apple (Annona Squamosa) Leaf Extract as an Additive Surfactant in a Liquid Eco-

Detergent
PEREZ, MARY YVONNE B.
TORRES, MATT ANGEL R.
First City Providential College
Senior High School Department

Abstract. Linear Alkylbenzene Sulfonate (LAS) surfactants are the active components frequently employed in detergents (S&P Global, 2021).
However, it has been discovered that mostly linear alkylbenzene sulfonates (LAS) have negative effects on aquatic and terrestrial ecosystems (B
Petrie., et al. 2015; Zhu, FJ, et al. 2018). Therefore, this study aimed to determine if the ethanolic extract of the leaves of sugar apple (A. squamosa)
is capable as a plant-based surfactant and as an antibacterial. This was established by testing the formulation of the liquid eco-detergent incorporated
with the ethanolic extract of the leaves of sugar apple (A. squamosa) subjected to the standards of the SNI 06-0475-1996, a document from Indonesia
that states the standard formula and characteristics of a liquid eco detergent, in which the standard values were given in determining if a liquid
detergent is safe based on the following: pH value, density and anti-microbial activity. The findings of this study revealed that the ethanolic leaf
extract of sugar apple (A. squamosa) is positive for saponins. Because of these findings, the researchers proceeded in manufacturing the product and
tested it afterward. The researchers found that the pH level of the samples indicates that the product is safe for direct human skin contact since the
human skin only tolerates a pH value of 6-8. After the density test was conducted, it was found that the density of the products is in accordance with
the density standards of SNI 06-0475-1996. Lastly, the product has shown antimicrobial properties against Staphylococcus aureus, a gram-positive
bacteria. Lastly, the means of three separate samples were examined using the paired sample t-test. The analysis' findings imply that there is no
significant difference between the first two samples' means and the second and third samples' means. The effect sizes, however, suggest that there
might be slight variations in these samples' means that are not statistically significant. To fully understand the nature of these disparities, more
investigation or data gathering may be required.

Keywords: Sugar Apple (Annona Squamosa) and its Leaf Extract, Eco-detergent, Preliminary Analysis, pH value, Density Test, Anti-Microbial Test

1. Introduction
A detergent is an organic material or mixture that contains soaps and/or surfactants and is used in washing and cleaning procedures (Health
and Safety Authority, 2023). Linear Alkylbenzene Sulfonate (LAS) surfactants, which are substances with two hydrophobic (lipophilic) and
hydrophilic (lipophobic) groups as well as detergency qualities, are the active components frequently employed in detergents (S&P Global, 2021).
However, it has been discovered that mostly linear alkylbenzene sulfonates (LAS) have negative effects on aquatic and terrestrial ecosystems in terms
of biochemistry, pathology, physiology, and other factors (B Petrie., et al. 2015; Zhu, FJ, et al. 2018).
Sugar apple (Annona s.) leaves contain a variety of chemical substances. This includes the following: phenolics, annonaceous acetogenins,
saponins, flavonoids, alkaloids, glycosides, alkaloids, steroids, and terpenoids (M Gowdhami., et al, 2015). Phenolics, in which the Sugar apple
(Annona s.) leaves contain, are used as disinfectants in household cleaners (Chemical Safety Facts.org, 2020). It also has been found that saponin-rich
plants are widely disseminated in nature, and they have significant promise for replacing hazardous synthetic surfactants in a range of current
commercial goods. These saponins also have outstanding surface and biological activity (S Rai., et al, 2021). Given the following findings, this study
aimed to produce a liquid eco-detergent incorporated with the extract of the leaves of A. Squamosa as a surfactant and an antibacterial. The goal of
this study is significant considering that synthetic detergents contain substances that lead to unfavorable results to the environment. These chemicals
are not only harmful to the environment, but they are also dangerous to the individuals that live in them since many of them are poisonous or can
cause illnesses or ailments.

Statement of the Problem

This study aims to test the efficacy of sugar apple (A. squamosa) leaf extract as an additive surfactant in a liquid eco-detergent by
conducting experimental research during the first and second semester of School Year 2022-2023.
Specifically, this investigation endeavors to answer the following sub-problems:
1. As a preliminary analysis, what are the phytochemical test results on the ethanolic leaf extracts of sugar apple (A. squamosa)?
2. What is the efficacy of sugar apple (A. squamosa) leaf extract as a surfactant in a liquid eco-detergent in terms of the following:
A. pH Value
B. Density
C. Anti-microbial Activity
3. What are the significant differences (if any) between the concentrations of sugar apple (A. squamosa) leaf extract as an additive
surfactant in a liquid eco-detergent?

2. Methodology

Research Design
The study follows a quasi-experimental research design in which the independent variable is the liquid-eco detergent based from the
ethanolic leaf extract of sugar apple (A. squamosa). Whereas the dependent variables will be the following: pH value, density and anti-microbial
activity.

Research Method Used

 Fig 1. A Schematic Diagram of the Research Method Used

Data Gathering Procedure

To gather the data for this study, the following are the procedures.
The ethanolic extraction method of the sugar apple (A. squamosa) leaves were based on El-Chaghaby’s study in 2018. The fresh leaves
acquired were thoroughly washed using running water and were air dried in direct sunlight for 5 days in humid conditions and the leaves were put
indoors whenever rainfall is imminent to avoid contamination. After 5 days of air drying, the leaves were then pulverised using a dry and a sterile
Imarflex kitchen blender. Subsequently, 6 grams of the pulverised leaves were taken and macerated in an incorporated 50 grams of distilled water and
50 grams ethanol for 72 hours in constant room temperature. After maceration, the leaf extract was centrifuged at 3000 rpm for 15 minutes and soon
filtered using a filter paper.
Seweryn & Wasilewski in 2019 successfully formulated an eco-liquid detergent with the inclusion of a plant extract. First, the following
ingredients were mixed with distilled water in a room temperature: 8 mL of sodium coco sulphate and 1 mL of cocamidopropyl betaine as a
surfactant, 0.001 mL of plant-based color and 0.005 mL of phenoxyethanol as an alternative preservative. Subsequently, the desired concentration
was added; the measurement of the distilled water will vary depending on the concentration of the leaf extract (5 mL, 10mL and 15 mL). The
viscosity of the formula must be similar to the viscosity of water. The desired measurement of the final product per sample is 100 mL.

Data Gathering Instruments

The results of this study will be subjected to the standards of the SNI 06-0475-1996. The following are the values that the variables should
attain in order to comply with the standards of the SNI 06-0475-1996, a document from Indonesia that states the standard formula and characteristics
of a liquid eco detergent: the pH value of the liquid eco-detergent must range from 6-8. Its density must range from 1.0-1.2 g/mL, its anti-microbial
activity criteria based on the zone of inhibition were as follows: 5-10 mm, very weak; 10-20 mm, moderate; 20 mm or more, very strong. Lastly, the
phenol coefficient value must be above 1. Any values that are less than or greater than the standard values that have been stated are considered as a
fail outcome in considering the standards of SNI 06-0475-1996.

Statistical Treatment of Data

After acquiring the necessary data, the researchers analyzed the obtained data using a statistical treatment. The statistical analysis of this
study was done using Paired T-Test with 5% level of significance, also known as the dependent sample t-test, is a statistical process for determining
if the mean difference between two groups of observations is zero. Each subject or entity is measured twice ia paired sample t-test, resulting in pairs
of observations. Jamovi statistics software was used and the formula for paired sample t test.

3. Results
1. As a preliminary analysis, what are the phytochemical test results
on the ethanolic leaf extracts of sugar apple (A. squamosa)?
Table 1. Phytochemical Test Results for Saponins
saponins.
2. What is the efficacy of sugar apple (A. squamosa) leaf extract as a
surfactant in a liquid eco-detergent in terms of the following:

Date: February 17, 2023 2.1. pH value

Table 2. pH Value Test Results

Date: February 17, 2023

Sample 1 Sample 2 Sample 3

Fig 2. Phytochemical Test Results for Saponins

Observation
After conducting the
phytochemical test, the
secondary metabolites of the
sugar apple (A. squamosa) leaf
extract in the form of saponins
was tested. The researchers
have observed that the foam of
the height formed is 3
centimeters.
Inference
The result of this experiment
indicates that the ethanolic leaf
extract of sugar apple (A. Fig 3.1. pH Value Fig 3.2. pH Value Fig 3.3. pH Value
squamosa) is positive for Test Results Test Results Test Results
saponins since according to (Sample 1) (Sample 2) (Sample 3)
Tunjungari in 2022, the Observation Inference
formation of foam with a height After conducting the pH level The result of this experiment
of 1-10 centimeter indicates that test, the ph value of the indicates that the product
the sample is positive for
 products with different samples are safe for direct
concentrations were tested. The human skin contact since the Table 4.1. Paired T-Test results of Sample 1 and Sample 2
researchers have observed that human skin only tolerates a pH
the pH value of the three value of 6-8. Moreover, in Paired T-Test Results of Sample 1 and Sample 2
Paired Samples T – Test
samples were the following: 6 testing the pH value of the statistic df p Effect
(5%), 8 (10%) and 6 (15%). product, the standard pH value Size
in according to SNI 06-0475- Sample 1 Sample 2 Student’s -1.31 8.00 0.226 Cohen’s -0.437
t d
1996 must be 6-8. Descriptives
N Mean Median SD SE
Sample 1 9 3.04 1.15 2.84 0.947
2.2. Density Sample 2 9 5.02 7.00 4.59 1.529
Table 3. Density Test Results

Date: February 17, 2023
Sample 1 Sample 2
Fig 4.1. Density Fig 4.2. Density
Test Results Test Results
(Sample 1) (Sample 2)
Observation
After weighing Sample 2 and
measuring its volume in terms
of mililiters, the researchers
have calculated its density and
observed that the density of the
sample products are as follows:
1g/ml (5%), 0.95g/mL (10%)
and 1g/mL (15%).
2.3. Antimicrobial Activity
Sample 3
Fig 4.2. Density
Test Results
(Sample 2)
Inference
The standards value of a liquid
eco-detergent’s density
according to the SNI 06-0475-
1996 is ranges from 1.0g/mL to
1.2g/mL. The results of this
experiment indicate that the
first and third sample act in
accordance with the density
standards of SNI 06-0475-1996,
however, the second sample has
a slightly low density which is
0.95g/mL.
The effect size is a measure of the magnitude of the
difference between the two sample means. A negative effect size
indicates that sample 1 has a smaller mean than sample 2. In this
case, the effect size is -0.437, which indicates that the difference
between the means is moderate. Overall, based on the results of the
paired sample t-test, we can conclude that there is no significant
difference between the means of sample 1 and sample 2. However,
the effect size suggests that there is a moderate difference between
the means, which may be of practical significance. However, in this
case, the p-value is 0.226, which is greater than 0.05. This means that
we do not have enough evidence to reject the null hypothesis.
Table 4.2. Paired T-Test results of Sample 1 and Sample 3
Paired T-Test Results of Sample 1 and Sample 3
Paired Samples T – Test
statistic df p Effect
Size
Sample 1 Sample 3 Student’s -0.151 8.00 0.884 Cohen’s -0.0504
t d
Descriptives
N Mean Median SD SE
Sample 1 9 3.04 1.15 2.84 0.947
Sample 3 9 3.39 1.00 5.74 1.912
The effect size is a measure of the magnitude of the
difference between the two sample means. A negative effect size
indicates that sample 1 has a smaller mean than sample 3. In this
case, the effect size is -0.0504, which indicates that the difference
between the means is very small.
Overall, based on the results of the paired sample t-test, the
researchers have concluded that there is no significant difference
between the means of the two samples. This is supported by the high
p-value and small effect size. Therefore, the researchers cannot reject
the null hypothesis that the means of the two samples are equal.
However, In this case the p-value is 0.884, which is much greater
than 0.05. This means that we do not have enough evidence to reject
the null hypothesis.

Table 4.2. Paired T-Test results of Sample 2 and Sample 3

Paired T-Test Results of Sample 2 and Sample 3

Paired Samples T – Test
statistic df p Effect
Size
Sample 2 Sample 3 Student’s 1.43 8.00 0.191 Cohen’s 0.477
t d
Descriptives
N Mean Median SD SE
Sample 2 9 5.24 7.00 4.74 1.58
Sample 3 9 3.28 1.00 5.80 1.93

The effect size is a measure of the magnitude of the

Fig 5. Laboratory Test Results for the Antimicrobial Activity
3. What are the significant differences (if any) between the
concentrations of sugar apple (A. squamosa) leaf extract as an
additive surfactant in a liquid eco-detergent?
difference between the two sample means. A positive effect size
indicates that sample 2 has a larger mean than sample 3. In this case,
the effect size is 0.477, which indicates that the difference between
the means is moderate. Overall, based on the results of the paired
sample t-test, the researchers have concluded that there is not enough
evidence to suggest that there is a significant difference between the
means of the two samples. This is supported by the p-value being
greater than 0.05, which indicates that the difference between the
means could be due to chance. However, the moderate effect size nature of the difference between the two samples. However, in this
suggests that there may be a meaningful difference between the case, the p-value is 0.191, which is greater than 0.05. This means that
means that is not statistically significant. It may be useful to gather we do not have enough evidence to reject the null hypothesis.
additional data or conduct further analysis to better understand the

4. Conclusion

1. This study aims to formulate an eco-liquid detergent with the inclusion of a plant extract. The raw material in this study, the ethanolic
leaf extract, is used as a plant-based surfactant and as an antibacterial, therefore, the presence of saponins is important as it has an outstanding surface
and biological activity (Rai, S., et al, 2021). The result of this experiment indicates that the ethanolic leaf extract of sugar apple (A. squamosa) is
positive for saponins since according to the SNI 06-0475-1996, the formation of foam with a height of 1-10 centimeter indicates that the sample is
positive for saponins.

2. The efficacy of sugar apple (A. squamosa) leaf extract as an additive surfactant in a liquid eco-detergent was tested in terms of the
following: pH value, density, and anti-microbial activity. The results for the pH level test indicate that the product is safe for direct human skin
contact since the human skin only tolerates a pH value of 6-8. Moreover, in testing the pH value of the product, the standard pH value according to
SNI 06-0475-1996 must be 6-8. Additionally, in conducting the density test, the standards value of a liquid eco-detergent’s density according to the
SNI 06-0475-1996 ranges from 1.0g/mL to 1.2g/mL. The results of this experiment indicate that the first and third sample act in accordance with the
density standards of SNI 06-0475-1996, however, the second sample has a slightly low density which is 0.95g/mL. Lastly, amongst the three product
samples, the second (10%) and the third (15%) samples showed antimicrobial activity against S.aureus, a positive-gram bacteria.

3. After analyzing the three pairs of paired t-tests, the researchers have concluded that there is a significant difference between the means of
sample 1 and sample 2, while there is not enough evidence to suggest a significant difference between the means of the other two pairs of samples.

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