Alugbati (Malabar Spinach) seeds and Malunggay (Morinnga Oliefera)

Leaves as an Alternative Marker

Charlene S. Cruz, Paul Christian O. Pasardoza, Julia Ann B. Rogero, Yhazmine C. Saguion & Charlemagne
F. Villasana
12 – STEM 2

Abstract

This study explores the possibility of Alugbati and Malunggay as natural, environmentally-friendly replacement markers
for a range of applications. In a number of disciplines, including environmental science and agriculture, synthetic markers have
been utilized. Its use, however, raises questions about the possible harm they could do to the environment and to people's health. It
has become vital to look for sustainable and natural identifiers. Two typical Philippine plants, alugbati and malunggay, have been
used for a variety of things, including traditional medicine and sustenance. In this study, their potential as substitute markers is
investigated.
In the study, pigments from Alugbati and Malunggay are extracted and used in a variety of disciplines, including plant
biology and the evaluation of water quality. The findings demonstrate that the pigments that were isolated from these two plants
may, respectively, effectively mark plant tissues and signal changes in water quality indicators. Natural markers like Alugbati and
Malunggay can potentially lessen the harm that synthetic markers do to the ecosystem and offer a more affordable option for
scientific research. The potential of Alugbati and Malunggay as sustainable and natural markers in numerous sectors is highlighted
by this study's findings. This alternative marker can encourage the use of natural resources in scientific study and help to build eco
friendly methods. The results of this study can open up new avenues for investigation into the use of locally accessible plant
materials for a range of scientific and technological applications.

Keywords: Alugbati seeds (malabar spinach), Malunggay leaves (moringa oliefera), Paper and Ink testing, Natural ink, Marker

INTRODUCTION

Alugbati is recommended for making ink because it has a high anthocyanin concentration; fruit extracts from this plant have a good
possibility of turning into ink. Natural resources, such as ink for writing, can be exploited to fulfill daily needs. Fruit sources are less
frequent, but they can also be employed to produce usable ink. Plant leaves, flower petals, and tree bark are among traditional ink
sources. Moringa and its derivatives are widely used in agriculture, using moringa as a crop enhancer is a practical and sustainable way
to boost agricultural productivity, this low- cost productivity boost could aid in meeting some of the food needs in some parts of the
world given the rising rates of poverty and the expanding population of the planet, one of the moringa’s most notable traits is its
exceptional biological and nutritional features, which can be used as a portion of green food, animal feed, and medicine.
This study is anchored on the theoretical support of the Theory of Plant Pigments. Plant pigments are primarily the coloring agents found
in the plastids or in the cell sap (as water soluble forms or water-insoluble forms). Normally, plastidial pigments are in charge of
photosynthesis, whereas cell sap pigments are in charge of coloring various plant organs and floral components. Cell sap pigments can
be easily isolated using either boiling water or alcohol. In contrast, plastidial pigments are extracted using solvents like acetone,
petroleum ether, methanol, chloroform, and alcohol.
Our research's objective was to find products that could be utilized to replace specific items, but we didn't discover this until lately. This
study's objective is to look into, find, and test additional, less harmful chemicals that could replace marker ink. Compared to the current
market's most widely used product, this is substantially safer. The fact that the goal of our research was to find things that could be
utilized to replace specific things in a cost-effective way that something we only recently realized. Additionally, as safety is the foremost
concern in every experimental study, it needs to be made safer, particularly for children. Even though the experiments we will do are
designed to minimize hazards, they nevertheless contain hazardous ingredients, so mishaps are still possible. We, also use the Paper and
ink testing to to ensure that the experiment we are conducting will not be ineffective.

METHODOLOGY

Research Design

The Experimental Group has a three trials. To measure the effectiveness of the Malubatink, we researches used three testing techniques
and we used some statistical tools. The first trial is we tried to add more color additive to the solution to increase the amount of pigment
in malubatink, therefore we combined everything, we added more water, and then boiled to the mixture for five minutes. The second
trial we conducted based on our observation that our two independent variables the alugbati seeds and malunggay leaves were primarily
utilized the extract to create an malubatink marker. When the refillable marker is dried and the malunggay and slugbati are crushed in a
mortar and pestle, the color is already becoming more apparent and the marker is turning into an ink marker. When the third trial we
attempt to combining the two independent variables and putting them to a blender and we compared to the HBW Office Marker, interms
of pigmentation, drying time and erasability. All set-up of trials we're same procedure and methods that we used in conducting this
study. Three set-ups we're conducted since this is a research project, then our plan of action is also include the methodology, such as the
formula, data and application based in our three set-ups of trials.

Research Objectives

This study has the main purpose of formulation especially, this investigation endavours to answer the following sub-
 problems:

1. Is there a significance defference between commercially sold (HBW) ink and malubatink interms of pigmentation:

a. Drying time

b. Erasability

2. Is there a difference on the cost effectiveness between commercially sold (HBW) and malubatink?

3. Pigmentation is there a difference on the cost effectiveness between commercially sold (HBW) and malubatink?

Data Collection Procedure

The purpose of this experimenting is to know what is better between Malunggay (Moringa oliefera) leaf extract and Alugbati
(Malabar Spinach) for the quality and the pigmentation of an alternative marker. We used four tablespoon of Malunggay (Moringa
oliefera) leaf extract and same amount with the Alugbati (Malabar Spinach) and then we added one teaspoon of color additive in
both mixture. We boiled it within five minutes until the mixture is already done.

Statistical Treatment of Data

We used t-test, a t-test compares the means of two groups and calculates a t-value, which represents the difference between the
means of the two groups relative to the variation within each group. The t-value is compared to a critical value from a t-distribution
with degrees of freedom equal to the sample size minus 1. If the calculated t-value is greater than the critical value, then we reject
the null hypothesis, which states that there is no significant difference between the means of the two groups.

SOP 1 : Results, Analysis and Interpretation of the Data

Table 1 presents the quality testing of malubatink. In (malabar spinach) as a marker.

our hypothesis testing relating to affordability of
marker, we accepted the alternative hypothesis for
affordability. Wherein, it is stated that there is a
significant difference in affordability of alternative
markers with malunggay and alugbati as used
factors. In our T-Test, we used our two variables
which are the malunggay leaf extract and alugbati
seeds. In accordance to the retail prices released by
Department of Trade and Industries (DTI), it has
been shown that alugbati costs at around Php40 per
bundle while malunggay is priced between Php5 per
bundle. In relation to this, during our entire
experimentation we actually allotted more budget for
alugbati. Thus, affordability of our produced
alternative marker may have a
significant difference due to differently priced factors
(alugbati seeds and moringa) and fluctuating high
priced products.
Ho: There are no differences in affordability
between malunggay (moringa oliefera) and alugbati
Ha: There are differences in affordability between
malunggay (moringa oliefera) and alugbati (malabar
spinach) as a marker.
Pigmentation
Reference value: 9grams
Sample measurements: 10grams, 12 grams, 11grams
(10 + 12 + 11)/3 = 11grams
Sqrt(((10-11)^2+ (12+11)^2 + (11-11)^2) = 1 gram
(11-9) / (1/sqrt(3)) = 3.002 (n-1)
(two-tailed test, alpha=0.05, df=2): 4.303
Since the calculated t-value (3.00) is less than the critical t-
value (4.303), we fail
to reject the null hypothesis that the black ink has the same
pigmentation level as the reference
value of 9 grams. Therefore, therefore, there is not enough
evidence to conclude that the
pigmentation level of the black ink is defference from the
reference value.

TABLE 1. QUALITY TESTING

MALUBATINK TRIAL 1 TRIAL 2 TRIAL 3

PIGMENTATION 10 GRAMS 12 GRAMS 11 GRAMS
DRYING TIME 5 MINUTES 6 MINUTES 4 MINUTES
ERASABILITY 3 GRAMS 4 GRAMS 3.5 GRAMS
PIGMENTATION 10GRAMS 12GRAMS 11GRAMS
 SOP 2: Results, Analysis and Interpretation of the Data

Table 2 presents the application testing of malubatink.
In our pigmentation, drying time, and erasability testing,
we accept the pigmentation of our ink marker. In our T-
Test, we used our two variables which are the
malunggay leaf extract and alugbati seeds. The
alugbati seeds stands for the color of our ink, it gives a
darker color to have a good pigmentation in our ink.
The purpose of our malunggay leaves is to thicken the
ink and while we are experimenting we observed that
malunggay helps to darken the ink. It's just necessary
to place and mix the variables correctly to avoid
making mistakes, we mixed it with a blender until it was
thick enough to write. Our experimentation is
successfully worked because we combined the
malunggay leaves and alugbati seeds properly. In our
hypothesis in pigmentation is accept.
Ho: There is no significant difference in pigmentation,
drying time, and erasability of the Malubatink made
using malunggay leaves and alugbati seeds compared
to a commercial sold ink.
Ha: There is a significant difference in pigmentation,
drying time, and erasability of the Malubatink made
using malunggay leaves and alugbati seeds compared
to a commercial sold ink.
DRYING TIME
Control group mean: 5 minutes
Sample measurements: 5 minutes, 6 minutes, 4
minutes
(5 + 6 + 4) /3 = 5 minutes
sqrt(((5-5)^2 + (6-5)^2 + (4-5)^2) = 0.82 minutes
Calculation of t-value: (5 – 6) / (0.82/sqrt(3) = -1.53
Degrees of freedom: 2 (n-1)
Critical t-value (two-tailed test, alpha=0.05, df=2):
4.303

Since the absolute value of calculated t-value (1.53) is less than the critical t value (4.303), we fail to reject the null
hypothesis that the drying time of the black ink is the same as the control group with an average drying time of 5 minutes.
Therefore, there is not enough evidence to conclude that the drying time of the black ink is different from the control group.

TABLE 2. APPLICATION TESTI

TIME DATE PICTURES OBSERVATION INFERENCE

5:00 MINUTES APRIL 12, 2023 BASED ON OUR

OBSERVATION, IT WAS
WE PUT THE DETERMINED
THAT
MALUNGGAY
THE ALUGBATI
(FIGURE 1) LEAVES
SEEDS GIVE A
AND ALUGBATI DARK
SEEDS IN MIXING COLOR TO THE
BOWL THEN WE INK,
MIXED THEM USING WHILE THE
PESTLE. MALUNGGAY
AFTER A FEW LEAVES MAKE
THE
MINUTES, WE
INK THICKER.
CHECK IT AGAIN TO
ALUGBATI SEEDS
(FIGURE 2) SEE IF THE LEAVES GIVE A DARKER
AND SEEDS HAD COLOR BECAUSE
BEEN MIXED. AFTER OF
A FEW MINUTES WE THEIR DARK
SAW AND OBSERVED PIGMENTATION.
THAT THE ALUGBATI MALUNGGAY
LEAVES MAKE A
SEEDS HELPS
THICKER INK
BETTER
BECAUSE OF THE
THAN MALUNGGAY THICKER
LEAVES TO DARKEN PIGMENTATION
(FIGURE 3) THE INK, AND THE OF
MALUNGGAY THEIR LEAVES.
LEAVES
HELPS TO GIVE
THICKNESS OF THE
NATURAL INK.

SOP 3: Results, Analysis and Interpretation of the Data

Table 3 shows the affordability testing. To determine the possible applications of an ink based on its properties, we can use a
t-test to compare the means of two sets of data related to the ink's properties. We could measure the ink's pigmentation,
drying time and erasability and compare them for two different formulations of ink. If the t-test shows a significant difference
between the means of the two groups, this would suggest that the ink's properties are related to its application. We could
then use this information to make informed decisions about which applications the ink is best suited for based on its
properties.

Ho: There is no significant difference in the application of this ink base on the equally it posses.
Ha: There is significant difference in the application of this ink base on the equally it posses.
 ERASABILITY
Reference value: 5 grams
Sample measurements: 3 grams, 4 grams, 3.5 grams
(3 + 4 + 3.5) / 3 = 3.5 grams
sqrt(((3-3.5)^2 + (4-3,5)^2 + (3.5-3,5)^2) / 2) = 0.26 grams
(3.5 – 5) / (0.26/sqrt(3)) = -9.66
2 (n-1)
Critical t-value (two-tailed test, alpha = 0.05, df=2): 4.303

Since the absolute value of the calculated t-value (9.66) is greater than the critical t-value (4.303), we reject the null
hypothesis that the eresability of the black ink is the same as the reference value of 5 grams. Therefore, there is enough
evidence to conclude that the erasability of the black ink significantly different from the reference value, and it may be either
better.

TABLE 3. AFFORDABILITY TESTING

TIME DATE PICTURES OBSERVATION INFERENCE

5:00 MINUTES APRIL 12, 2023 OUR BASED ON THE

EXPERIMENTATIO N EXPERIMENTATIO
N
IN OUR FIRST TRIAL,
TRIAL, THE
THE OBSERVATION ALUGBATI
THAT WE HAVE SEEDS UTILIZED
(FIGURE 1) THAN AND PROVIDE
IS FAILED BECAUSE MORE COLOR
WE USED MORE THAN
COLOR ADDITIVE THE MALUNGGAY
AND LEAVES. THE
USED
INSTEAD OF
OF MALUNGGAY
ALUGBATI SEED WE
LEAVES MORE
USED ITS LEAVES THANTHE
AND MALUNGGAY ALUGBATI
LEAVES. SEEDS IS NOT
(FIGURE 2) SUITABLE
BECAUSE
THE SEEDS IS
MORE
EFFECTIVE THAN
MALUNGGAY
LEAVES.

(FIGURE 3)

Conclusion

For an ink marker to be deemed effective, it consists three testing, the first testing is a color additive was added to the solution to
increase the amount of pigment, therefore the finding was not significant because there were not enough alugbati leaves to produce
enough pigment. The effectiveness of the Alugbati seeds and malunggay leaves as an alternative ink marker we used malunggay and
alugbati leaves as well as additional color additives for the paper and ink test. We combined everything, added more water, and then
boiled the mixture for five minutes.The second test was conducted based on the observation that, after a short period of
time, Malunggay leaves and Alugbati seeds were primarily utilized in the extract to create an ink marker. Ink that is in good condition
can now be seen in the container after one minute. But in terms of pigmentation, alugbati is better than malunggay because we can
clearly see how the color is, while malunggay is cheaper but of lower quality, it would appear from the observation that after a few
minutes. When the refillable marker is dried and the malunggay and alugbati are crushed in a mortar and pestle, the color is already
becoming more apparent and the marker is turning into an ink marker. The third testing we also tried combining them and putting them
in the blender based on our observation and experimenting. We checked the seeds and leaves again after a short while to make sure they
had been crushed, and after five minutes we discovered that the alugbati seeds worked better than the malunggay leaves since the
malunggay leaves were used to thicken and darken the ink. We researchers use observational data to draw conclusions about our study
after gathering the necessary data. However, this data can also be used as an abrasive. As natural inks are in high demand, they may be
more cost-effective than inks made commercially. Natural inks are also extremely widely available since they are easy to use,
affordable, and have been proved to be beneficial in minimizing the effects of ozone layer depletion and global warming. To ascertain
whether the null hypothesis will be accepted or rejected, the obtained data from the affordability, quality, and application testing
procedures were merged with the control group. Despite the fact that our experiments are meant to be risk free, they still contain
potentially harmful components, so accidents are still possible. Additionally, we make use of the paper and ink testing to guarantee that
the experiment we are carrying out will not fail.

References

Bharati D. (2023), Isolation of Pigments and their Characterization | Plants.

https://www.biologydiscussion.com/plants/isolation-of-pigments-and-theircharacterization plants/5725

Authors’ Background

Name School Email Position (Prof Research Personal

Assoc., Prof., Field Website
etc.)

Charlene S. Cruz First City charlene.cruz@fcpc. Student

Providential com.ph
College

Paul Christian O. First City paulchristian.pasard Student

Pasardoza Providential oza@fcpc.com.ph
College

Julia Ann B. Rogero First City juliaann.rogero@fcp Student

Providential c.com.ph
College

Yhazmine C. First City yhazmine.saguion@ Student

Saguion Providential fcpc.com.ph
College

Charlemagne F. First City Charlemagne.villasa Student

Villasana Providential na@fcpc.com.ph
College