1

Feasibility of Rambutan (Nephelium lappaceum)

Seeds as a Water Filter

Celesly C. Carza
Carla Arriane V. Gabat
Alexandra Fe C. Garcia

Abstract: This study presents the feasibility of Rambutan (Nephelium lappaceum) seeds as an affordable and
organic agent to treat water. To satisfy this study, four laboratory testings were conducted - (1) Phytochemical
Screening, (2) Experimentation, (3) Water Microbacteriology Analysis, and (4) Urine Analysis. Due to the
potential biocoagulant activity of the Rambutan samples proved by past researches, seeds then were crushed
thoroughly into powder followed by mixing it in an Erlenmeyer flask with 20mL distilled water to make the
Rambutan seed solution. The solution was then transferred into the beaker containing 500mL untreated water
then stirred well. After stirring, the treated water was left undisturbed for 1 hour in the laboratory to let the
flocs sit. When the particles and contaminants have settled to the bottom, the now treated water was carefully
transferred to another beaker using filter paper and glass funnel which creates the Rambutan seed filtered
water. The Phytochemical screening shows presence of Alkaloids which aids with its antioxidant and
antimicrobial activities and Tannins which mainly acts as color of the treated water which also is responsible for
bad odor and taste. Correlation between experimental group and pondwater is at 0.28 (moderately small
correlation). Using Analysis of Variance, statistics on significant difference is determined. This study supports
past studies on the potential and feasibility of Rambutan (Nephelium lappaceum) seeds to filter untreated
water.

Keywords: Rambutan, seed, filter, biocoagulant

INTRODUCTION
Water crisis is already ranked 4th as a
global risk in terms of impact according to the
World Economic Forum (2019). Furthermore,
according to the World Health Organization
(2015) and United Nations International
Children’s Emergency Fund (2015) one out of
nine people in the world have no access to safe
drinking water and one- third of the global
population doesn’t have access to toilets. This
calls for the need for action to search for
affordable and simple solutions (Okumu &
Natumanya, 2015). This is evident even here in
the Philippines. Around six million Filipinos still
practice open defecation and 20 million lack
access to basic sanitation facilities (UNICEF,
2017). With these unclean practices, they are
free to countless diseases. Being far away from
the sanitary help and safe drinking water, the
objectives of this study includes finding a way to
help the situation with an affordable and
organic solution.
Rambutan in the Philippines is in its
season from August to October (Malasig, 2018)
When Rambutan is in its season people can buy
it at a low price because of increase in demand.
According to the Bureau of Agricultural
Statistics (2010) the volume of Rambutan in the
Philippines is 12, 743.16 metric tons covering a
total area of 5, 743.08 hectares. With those big
numbers their corresponding seeds can be used
to help clean water especially when it is
abundant when rain and flood is prevalent in
the country.
Apparently, only a few studies were
found with subjects focusing on organic water
filters especially as Rambutan seeds as the
subject.
Theoretical Framework
Aquatic plants have the capability to
decrease water pollution level (Tano, Sawada &
Oyabu, 2006). It is just recently that there are
some studies and experiments on non-aquatic
plants which can also filter dirt in water. One of
those were the Moringa oleifera seeds. The
study of Robert Natumanya (2015) from

1

Makerere University College of Agricultural and
Environmental Sciences is at the forefront of a
research to enhance water purification by
embracing green technology of using moringa,
jackfruit and Java plum (jambula) seeds.
Another study by Beth Doerr et. al.
(2005) contains specific procedures on using
Moringa oleifera to filter water and lessen
levels of turbidity. In this study they resort to
simple methods wherein the contaminants
would clump together at the bottom of the
container along with the crushed seeds and are
then ready to be physically filtered using the
laboratory filter paper.
In order to clean water, water filters use
two different techniques to remove dirt
(Woodford, 2018). Physical filtration happens
when straining water to remove large or
obvious impurities. The other method of
filtering is chemical filtration wherein it involves
passing water through an active material in
order to remove impurities chemically, usually
impurities not seen by the naked eye, as they
pass through. In this study physical filtration is
employed by the use of a laboratory filter paper
on the other hand chemical filtration is
employed due to the potential biocoagulant
activity of the Rambutan (Nephelium
lappaceum) seed according to a research by
Abidin et. al. (2014). To add more from their
study the Rambutan seed coagulant exhibited
faster sedimentation time due to bigger flocs
formation.
The researchers pursued the use of
Sprague dawley rats because according to
Melina (2010) their genetic, biological and
behavior characteristics closely resemble those
of humans, and many symptoms of human
conditions can be replicated in mice and rats.
Furthermore, Kurien et. al. (2004) indicated
mice and rats are two of the primary
mammalian species used for the preclinical
evaluation of drugs and preregistration
evaluation of chemicals.
2
2
Since water is considered a chemical
substance as said by the Human Metabolome
Database (2019), the researchers secured
Sprague Dawley rats for urinalysis to test their
urine pH levels as urine acidification indicates
an intake of acid (de Vries EG, 1986). To add on
that, according to Piech and Wycislo (2018)
urinalysis is often considered to be an important
diagnostic test by veterinary specialist to
identify tract disorders and other diseases in
our excretory tracts. Also, according to Rango,
et. al. (2015), there is a chronic kidney disease
going around with unknown (“u”) cause which
is creating a public concern in the areas of Sri
Lanka. Prior to their research, some has
hypothesized a link with drinking water quality,
although studies are lacking.
Three comparable groups were used –
one experimental group, one positive control,
and one negative control. A similar study by
Wan (2018) used pond water as well as a
sample for filtration hence the negative control
pond water which were gathered in a fish pond
of Virgen Milagrosa University Foundation was
employed. According to Brenner (2018),
distilled water offers the best choice for use in
science projects is that it is inert, which means
little to nothing remains in the water after
distilling that is why the researchers chose
commercialized distilled water as the positive
control. The plant sample Rambutan
(Nephelium lappaceum) seeds filtered water
was the experimental group.
Conceptual Framework
The paradigm of the study focused on
the feasibility of Rambutan (Nephelium
lappaceum) seeds as a water filter. The seeds
have undergone certain processes in which they
are applied and tested to create the output
which was the Rambutan seed as a water filter.
The Figure 1 on the following page
shows the Research Paradigm.

Figure 1. Research Paradigm
Objectives
This research study was conducted to
determine the feasibility of Rambutan
(Nephelium lappaceum) seeds as a water filter.
Specifically, it sought to determine (1)
the active constituents of Rambutan
(Nephelium lappaceum) seeds; (2) the
microbacterium effect of the experimental
group on test subjects; (3) the difference
between the experimental Rambutan
(Nephelium lappaceum) seeds groups and the
control groups.
Scope and Delimitations
The study focused and is limited only in
determining the feasibility of Rambutan
(Nephelium lappaceum) seeds as a water filter.
The Rambutan seeds were gathered in San
Carlos City, Pangasinan. The study used the
pond water gathered in the Virgen Milagrosa
University Foundation Pond as a negative
control while using a commercialized distilled
water as the positive control. Eighteen (18)
Sprague dawley rats bought from Agtas, San
Juan, San Carlos City, Pangasinan were
employed and used pH levels as parameter for
feasibility.
To satisfy this study 4 laboratory
testings were conducted - (1) Phytochemical
3
3
Screening, (2) Experimentation, (3) Water
Analysis, and (4) Urine Analysis.
The Phytochemical Screening as well as
the experimentations were carried out in the
College of Pharmacy Building of Virgen
Milagrosa University Foundation, San Carlos
City, Pangasinan while water analysis was
conducted by the Microbiology laboratory of
the Medical Laboratory Sciences Department,
VMUF, San Carlos City, Pangasinan. Urine
Analysis was held in AGS Diagnostics Laboratory
located at Rizal Ave., San Carlos City,
Pangasinan in the year 2018.
As an additional testing for analysis of
samples, only Water Bacteriology Analysis were
conducted to test for Coliform. Further
processes or tests were not conducted as well
as further biological effects on the rats were not
determined.
MATERIALS AND METHODS
Research Design
This research utilized an experimental
research design. This was employed because
the study performed experiments and
processes for testing of hypotheses to find

conclusions to support the feasibility of the
Rambutan (Nephelium lappaceum) seeds as a
water filter and its relationship with the other
controlled groups.
Collection and Preparation of Sample
Upon collecting the Rambutan
(Nephelium lappaceum) seeds they were
washed with the use of distilled water inside
the VMUF College of Pharmacy laboratory. In
preparation for the extraction, with the use of
mortar and pestle, the rambutan seeds were
crushed into smaller fragments.
The pieces were placed in an
Erlenmeyer flask. While in a graduated cylinder,
600 ml of ethanol was measured and poured
into the Erlenmeyer flask with the crushed
Rambutan seeds.
After 3 days, the mixture was heated at
340 ºC for an hour using the hot plate and
metal tray. Average amount of water was
poured on the metal tray and the Erlenmeyer
flask was placed atop on it. When the mixture
cooled, it was filtered using filter paper. More
ethanol was added to the mixture and was
filtered until 500 ml was gained.
The pure rambutan seed extract was
obtained through water bath process using the
evaporating dish, hot plate, and hot bath. The
500 ml was heated at 550 ºC until the ethanol
evaporated. The pure extract was transferred
to the petri dish.
Procedure of Phytochemical Screening
The procedure starts with Hot
maceration method using Soxhlet apparatus.
Freshly collected plant materials were air dried
and then coarsely powdered in a blender. 100 g
of the coarse powder was extracted
successively with 250 ml of various solvents
(ethanol, petroleum ether, chloroform,
methanol, water) in a Soxhlet apparatus for 24
h. All the extracts were filtered through No. 41
filter paper separately and all the extracts were
subjected to qualitative tests for the
identification of various phytochemical
constituents as per the standard procedures
(Brinda et al., 1981; Anonymous, 1990 and Lala,
1993). The extracts were concentrated in a
4
4
rotary evaporator. The concentrated extracts
were used for further analysis. It is then
followed by the preliminary Phytochemical
screening of different extracts. The chemical
tests for various phytoconstituents present in
the extracts were carried out (Brinda et. al.,
1981). Tests for Alkaloids, Anthraquinone
(Borntrager’s test), Catechin, Coumarin,
Flavonoid (Shindo’s test), Phenol, and Quinone
were conducted.
Experimental Procedure
Experimental procedures were based
on a similar study by Beth Doerr et. al. (2005)
which contains specific procedures on using
Moringa oleifera to filter water and lessen
levels of turbidity.
After collection and preparation of the
Rambutan (Nephelium lappaceum) seeds, they
determined the quantity needed based on
amount of water; in general, 3 seeds treated
500mL of water (one bottle). The seeds then
were crushed thoroughly with the use of mortar
and pestle an in order to obtain a fine powder,
they were sifted through a sieving machine. It is
then followed by mixing the seed powder in an
Erlenmeyer flask with 20mL distilled water to
make the Rambutan seed solution. The solution
was stirred for 1 minute to activate coagulant
properties.
Afterwards, the solution was then
transferred into the beaker containing 500mL
untreated water which was stirred rapidly for at
least 1 minute then slowly (15–20 rotations per
minute) for 5 minutes to ensure treatment.
After stirring, the treated water was left
undisturbed for 1 hour in the laboratory to let
the flocs sit. When the particles and
contaminates have settled to the bottom, the
treated water was carefully transferred to
another beaker using the filter paper and glass
funnel which creates the Rambutan seed
filtered water.
Procedure of Water Microbiology Analysis
The Water Microbiology Analysis was
conducted with the VMUF Microbiology
laboratory of the Medical Laboratory Sciences

Department. This analysis was done to
determine the E. coli presence among the
groups. According to Cabral (2010),
Microbiological water analysis is mainly based
on the concept of fecal indicator bacteria in
accordance with the background of this study
wherein open defecation is endemic.
a. Colony Count
Prepare 3 dilution 1:10, 1:100, and 1:
1,000 into 3 test tube. From 3 test tubes
pipette 9 mL of distilled water and place it
to each tube. Label 1:10, 1:100, and 1:
1,000. Pipette 1 mL of the sample then
place it to 1:10 tube from 1:10 pipette 1
mL then place it to 1:100 then pipette 1
mL from 1: 100 and transfer to 1:1,000.
From the last dilution pipette 1 mL and
place it to empty petri dish then pour the
melted Nutrient Agent. Incubate the plate
at 37℃ for 24 hours. After 24 hours of
incubation, count the number of colonies
present.
b. Test for Coliform (Presumptive Test)
Prepare Durham fermentation tube and
10 mL of Lactose broth. Place 1 mL of the
sample form 1:10, 1:100, and 1: 1,000.
Incubate for 24 hours to 48 hours.
Observe the production of gas then count
the colonies.
c. Confirm Test
From the presumptive test positive. Get
a loopful colony and then inoculate it to
Macconkey Agar by streaking. Incubate
the plate for 24 hours. After 24 hours,
observe the colony produce either lactose
(pink colony) or non-lactose fermenters
(white colony).
d. Completed Test (Biochemical Test) for
negative control
Urine Analysis (Application of Treatment)
This investigation was tested to eighteen
(18) Sprague dawley rats which were gathered
from Agtas, San Juan, San Carlos City,
Pangasinan. Before experimentations are done,
the rats were subjected to acclimatization for
seven (7) days.
5
5
Handling procedures were conducted
with the supervision of a laboratory personnel
and includes placing the rat on a surface it can
grasp. It is suitable surface is a wire cage top or
a towel. Applying a slight tension to the base of
the tail so the rat grasps onto the surface and
then gently but firmly place your free hand over
the shoulder and quickly grasp the scruff of the
neck close to the base of the skull between the
thumb and forefinger. Finally, restrain the tail
by the little finger.
The treatments, by the use of the ratio-
proportion method, were orally administered
by the use of a laboratory gavage. Three
readings were done to get results. For the first
reading the positive control distilled water was
administered to all rats. On the second reading
the negative control pond water was
administered followed by the third reading
where the experimental rambutan seeds
filtered water was administered.
For the collection of urine sample, the
urine was collected morning peak using
specimen urine containers after administration
of different experimental groupings
(subdivided) for Sprague dawley rats. Each rat
was lifted up while waiting for urine to come
out as it is directly poured into small test tubes
for reading using chemical urinalysis strips
(alpinum Prazision).
The method used for rat disposal was
the Physical Method (Cervical Dislocation)
under the supervision of an IACUC trained
Veterinarian.
After the data was gathered from urine
analysis, the researchers focused on the data
regarding the potential hydrogen (pH) level to
determine the relationship between
treatments.
Statistical Tools
For data analysis, the researchers used
the statistical technique named correlation
coefficient specifically the Pearson r. This was
done to determine the correlation between
 6

groups for identifying the more effective group.

Alongside the said tools, probability level of
(p<0.05) were considered significant. Through
the use of the IBM SPSS Statistics 23 and cross-
checking with Microsoft Excel 2016, the results
were computed.

RESULTS Phytochemical Screening was

conducted at VMUF College of Pharmacy
Table 1 Laboratory to cross- check with other
Active Constituents of Rambutan Seeds researches like Thitilertdecha, et. al. (2008).

Active Constituents Uses To know the microbacterium effect of

Alkaloids Alkaloids have several the experimental group on chosen test subjects,
important biological a Water Bacteriology Analysis was conducted.
activities as
antimicrobials, anti- Test for Coliform indicated that the
inflammatory, negative control contained Lactose Fermenters
antioxidants. (Abdel- and E. coli.
lateif KS et. al., 2016)
Tannins Tannins can cause a
yellow to brown cast in
water and may also
impart a taste and
odor. (Ashok, 2012)

Table 2
Statistical Results
Summarized Results of Water Bacteriology on Three Comparable Groups

Test Positive Exp. Group Negative Control

Control
Colony Count 1: 10 1: 10 1: 10
10 15 22

1:100 1:100 1:100

8 12 19

1: 1,000 1: 1,000 1: 1,000

7 10 18

Test for Coliform

Presumptive Test (MPN) 1: 10 1: 10 1: 10
10 15 22

1:100 1:100 1:100

8 12 19

1: 1,000 1: 1,000 1: 1,000

7 10 18
6
Confirm Test No Growth No Growth Lactose Fermenters
Completed Test Negative Negative E. Coli
Gram Stain No Microorganism Seen No Microorganism Seen Gram (-) Bacilli
 7

Table 3
Correlation Between Positive Control
And Experimental Group pH Levels
Rat No. Experimental Negative
pH Levels Group (x) Control
Rat No. Experimental Positive Control (y)
Group (x) (y) 1 6.5 5.0
2 6.5 6.0
1 6.5 7 3 7.0 6.5
2 6.5 7 4 7.0 6.5
3 7.0 6.5 5 7.0 6.0
4 7.0 7 6 6.5 6.5
5 7.0 6.5 7 7.0 6.0
6 6.5 7 8 7.0 6.0
7 7.0 7 9 7.5 6.0
8 7.0 6.5 10 6.5 6.5
9 7.5 6.5 11 7.5 7.0
10 6.5 7 12 8.0 6.0
11 7.5 6.5 13 7.5 6.0
12 8.0 7 14 7.5 6.5
13 7.5 6.5 15 6.5 5.0
14 7.5 7 16 7.0 6.0
15 6.5 7 17 7.5 6.0
16 7.0 7 18 7.0 5.0
17 7.5 7 Pearson r Correlation Value
18 7.0 7 = 0.28779693 (Moderately Small)
Pearson r Correlation Value
= -0.4030342 (Moderately Small) On the other hand, Correlation value of
According to Khatoon et. al. (2013), Pearson's 0.28 between the Experimental Group and the
correlation coefficient (r) value is determined negative control pond water shows that
using correlation matrix to identify the highly although very small, the Rambutan seeds still is
correlated and interrelated water quality more effective than pond water.
parameters. The Correlation value of -0.4
between the positive control and the Rambutan Table 5 on the following page, ANOVA
seed filtered water indicates that the positive was tested at (p<0.05) level of significance. With
control group distilled water is still more the tabulated value of 27.73 being higher than
acceptable than the experimental group alone. the F -critical value of 3.18, the null hypothesis (
H 0), which states that there is no significant
difference between groups, is rejected. In terms
of urine pH levels after drinking each sample
groups, there is a significant difference.

Table 4
Correlation Between Experimental Group and
Negative Control

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Table 5
ANOVA between Three Comparable Groups
Source of Variation SS df MS F P-value F crit
Between Groups 10.5277778 2 5.263889 27.731707 5.3708 3.178799
Alkaloids
Within Groups have several important
9.68055556 51 Thitilertdecha, et. al. (2008) which says that
0.189815
biological activities as antimicrobials, anti- several potential antioxidant activities were
inflammatory,
Total
and antioxidants therefore the
20.2083333 53  
evaluated   in their study
 
about  the seed
Phytochemical screening that was conducted by extracts.
the researchers supports the study of
cause a yellow to brown cast in water and may
DISCUSSION
also affect a taste and odour (Ashok,
S
Upadhyaya, 2012). The results of the CONCLUSION
phytochemical screening and the Figure 2,
which shows the color of the Rambutan seeds After analysis and interpretation of all
filtered water (leftmost) against the other the gathered data, the researchers concluded
treatments, supports the said claim. It also that the Rambutan (Nephelium lappaceum)
supports Rohman (2017) that reports several seeds has a very small positive potential in
biological activities, which are beneficial to being a water filter to untreated water.
human health in rambutan fruit, namely Although its relationship with the
antioxidant, antibacterial, antidiabetic and commercialized distilled water is not as strong
anticancer. The screening also found out the due to a negative value (-0.40) in correlation
presence of tannins in the rambutan seeds. it still shows promise just as the study of Abidin,
Although these compounds are not a health et. al. (2014) resulted in. The continuous search
risk, they are aesthetically displeasing. for a solution on water crisis can start with
The microbiological analysis presents enhancing the ability of the seeds to be natural
that untreated water that is filtered with biocoagulants. Studies like this can lead to an
Rambutan (Nephelium lappaceum) seeds does affordable and organic solution.
not have e.coli and lactose fermenters.
Although
RECOMMENDATION
it does not prove the water to be a safe drinking
water the subject is still recommended to be
Based on all the findings and
used as a potential filter.
conclusion of the study, recommendations were
The results on Pearson r correlation
made. It is recommended that other family or
analysis sees that Rambutan seeds are indeed,
other parts of the Rambutan (Nephelium
although very small in strength, shows potential
lappaceum) fruit be tested as a possible water
in filtering untreated water without aid from
filter. Also to find ways in order to make the
already known agents in filtration. A study of
seeds’ shelf life longer. There are researches
Abidin, et. al. (2014) says the same on
that say the Rambutan peel has more alkaloids
Rambutan seeds potential to remove turbidity
that work as antioxidants and antimicrobials. If
although in their study, the use of alum is
not for water filtration, other researches that is
employed.
related on life or physical science is
The difference between the groups is
recommended to use Rambutan fruit as their
considered significant although that is in terms
main subject or focus because of its big volume
in possible microbacterium effects on the rats
production. Further tests or processes is
as they remove wastes. It can be said, along
recommended to be done. A thorough analysis
with the Pearson r analysis, that pond water
or testing on its microbacteriology content,
and the experimental group has stronger
toxicological tests, increase in number of set-
relationship in this study.

8
 9

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various high -quality laboratory equipment
within the local area, it is recommended to
reach out to well-known facilities to get even
more set of good data to analyse and interpret.
Further experiments and research on this title is
recommended to complete a possible future
product that can be created and ensure
maximum use of Rambutan seeds potential and
capability. Lastly, it is recommended to conduct
a research using this subject as a drinking water.

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