Mapa Institute of Technology

School of ChE-Chm-BE-MSE
Muralla St., Intramuros, Manila

Special Project: Quality analysis of drinking water from MIT

dispensers
Authors: Afable, Lee Raven S.; Banta, Mary Claire L.; Manucduc, Nehemia Joy P.; Pineda, Aaron Jesnikho M.; Yu, Mark Francis C.

Abstract
Potable water is water which is fit for consumption by humans and other animals. Water dispensers
around the campus can help students save money from buying bottled waters from time to time by just
refilling their water containers. Water may be naturally potable or it may need to be treated in order to be
safe. In either instance, the safety of water is assessed with quality analysis that tests for the presence of
potentially harmful contaminants in the water. Twenty three water dispensers in Mapua Institute of
Technology Intramuros specifically in the North, West and South building were tested through swabbing
and water sampling, hot and cold water were analyzed by Ultraviolet-visible spectroscopy (UV-Vis) and
Spectrophotometry to test the turbidity of the samples. In this experiment, there are 72 samples in total:
46 for the swabbing and 26 for the water samples. Nutrient agar was used in the swabbing process with
the ingredients of water, sodium chloride, beef extract, agar and peptone. As a result, the safest water
refilling station in the campus is located in the North Building 4 th floor and the most turbid samples are
located in the South building area.

Keywords: Potable water, contaminants, UV-Vis, Spectrophotometer, Turbidity

Introduction
Drinking water, also known as potable water or
improved drinking water, is safe enough for drinking
and food preparation. In 2012, 89% of people had
access to water suitable for drinking. Nearly 4 billion
had access to tap water while another 2.3 billion had
access to wells or public taps. 1.8 billion People still
use an unsafe drinking water source which may be
contaminated by feces. This can result in infectious
diarrhea such as cholera and typhoid among others.
Water is essential for life. The amount of drinking
water required is variable. It depends on physical
activity, age, health issues and environmental
conditions [1].
Typically in developed countries, tap water
meets drinking water quality standards, even though
only a small proportion is actually consumed or used in
food preparation. Other typical uses include washing,
toilets, and irrigation

[2]. Greywater may also be used

for toilets or irrigation. Its use for irrigation however

may be associated with risks. Water may also be
unacceptable due to levels of toxins or suspended
solids.
Reduction
of waterborne
diseases and
development of safe water resources is a major public
health goal in developing countries. Bottled water is

sold for public consumption in most parts of the world.

The word potable came into English from the Late
Latin potabilis, meaning drinkable [3].
Buying drinking water from refilling stations
has become the most common way of obtaining clean
and safe drinking water. There are now more than
20,000 water refilling stations in the country, where up
to 40% of households nationwide and 60% of Metro
Manila residents buy drinking water [3].
Proactivity is encouraged for consumers to get
the cleanest drinking water possible. As a research
study from the Paediatric Infectious Disease Society of
the Philippines puts it, Characteristics of the [water
refilling] stations such as the disinfection method,
frequency of monitoring, personnel hygiene, attire and
practices all affect the safety and potability of the
product water [4].
Turbidity of water is an optical property that
causes light to be scattered and absorbed, rather than
transmitted. The scattering of the light that passes
through a liquid is primarily caused by the suspended
solids. The higher the turbidity, the greater the amount
of scattered light. Even a very pure fluid will scatter
light to a certain degree, no solution will have zero

Spectrophotometry is a method to measure

how much a chemical substance absorbs light by
measuring the intensity of light as a beam of light
passes through sample solution. The basic principle is
that each compound absorbs or transmits light over a
certain range of wavelength. This measurement can
also be used to measure the amount of a known
chemical
substance.
Spectrophotometry
is
a
quantitative measurement of the reflection or
transmission properties of a material as function of
wavelength. Also, electromagnetic (EM) radiation may
be absorbed by matter that is responsible for the
spectrophotometric methods. The extent to which the
EM radiation is related to the nature of concentration
of absorbing material present in a sample as well as
the wavelength of the radiation employed [6].
There are twenty three water dispensers in
Mapua Intamuros that were tested through the process
of swabbing and water sampling. Hot and cold water
were collected and analyzed by the UV-Vis and
spectrophotometry. The objective of this experiment is
to determine where in the campus has the safest and
the cleanest water dispenser for students to know
where they could refill their bottled waters and jars.
Through these findings, we can addressed the results
to the Mapua administration on the current situation of
the water dispensers and in order for them to take an
action on this.

5. Make a barium sulfate solution for the control

sample with concentrations of: 0.01M, 0.005M,
0.003M, 0.002M, and 0.001M.
6. Before running the spectrophotometer, set its
wavelength to 600 and let it pass through a
blank sample (i.e. saline distilled water) placed
in the cuvette.
7. After running it with the blank sample, run the
spectrophotometer with the control sample as
well as the samples containing bacteria.
8. Plot the gathered data with y-axis as
absorbance and x-axis as its concentration.
9. From there calculate for the concentration of
the individual sample.

Results and Discussion

Table 1. Standard Values

Standard Concentration
(mg/ml)
BLANK

Absorbanse (AU)

0.001

0.220

0.002

0.574

0.003

0.745

0.005

1.204

0.010

1.920

3
Absorbance

turbidity. Turbidity is commonly treated using either a

settling or filtration process. Depending on the
application, chemical reagents, will be dosed into the
wastewater stream to increase the effectiveness of the
settling or filtration process. Potable water treatment
and municipal wastewater plants often remove
turbidity with a combination of sand filtration, settling
tanks and clarifiers [5].

y = 190.88x + 0.1091
R = 0.9761

2
1
0
0

Methodology

0.01

0.015

Concentration

STANDARD
3
Absorbance

1. Make a media for the bacteria (i.e. nutrient

broth)
2. Get a water sample from the two spouts (i.e.
hot and cold water outlet) of the dispenser
using a sterilized vial, fill the vials up to the
mark (ensure that the caps are well sealed)
3. Get a sample from the dispenser by swabbing
a sterilized cotton buds to the spouts of the
dispenser/s to be analyzed and mix the
swabbed cotton buds with the media in the
vial.
4. Culture the vials with different samples for two
to three days to make sure that the bacteria
have grown in the containers.

0.005

2
1
0
0

0.005

0.01

Concentration (mg/mL)
Figure 1. Concentration vs. Absorbance

0.015

1st Floor Swabs

Absorbance

The samples were tested for the concentration

of microbes present by the determination of turbidity.
This relationship is stated in the Beer-Lamberts Law.
The Beer-Lambert law relates the attenuation of light
to the properties of the material through which the
light is traveling, which includes the path length,
concentration of the solution, and intensity of the light
after passing through the medium. This concept was
used to create a graph for the standard. Since the path
length is constant, the graph is concentration vs
absorbance [7]. Table 1 shows the data plotted to
create the standard graph. This was used as a basis to
get the concentration of the samples for the other
results.

3
2

STANDARD

1st F (SC)

0
0

0.005

0.01

1st F (SH)

0.015

Concentration (mg/mL)
Figure 2. First floor swabbing results graph

Table 2. First Floor Swabbing Results

Dispe
nsers
A
(HOT)

Absor
banc
e
(AU)

S01A

0.560

S07A

S08A

S14A

S15A

1.712

0.565

1.096

0.038

Concen
tration
(mg/ml)

Dispe
nsers
B

Concen
tration
(mg/ml)

Dispe
nsers
A

1.318

0.00627

(HOT)

(COLD)
0.00239

0.00828

0.00242

0.00513

0.00000

S01B

S07B

S08B

S14B

S15B

Table 3. Second Floor Swabbing Results

Absor
bance
(AU)

1.328

1.374

1.790

1.477

Absorb
ance
(AU)

Conce
ntratio
n

Dispe
nsers
B

(mg/ml)

(COLD)

S02A

0.620

0.00270

S11A

0.706

S16A

S23A

Absor
bance
(AU)

Conce
ntratio
n
(mg/ml)

S02B

1.892

0.00920

0.00314

S11B

1.585

0.00763

0.007

0.00000

S16B

1.624

0.00783

1.237

0.00585

S23B

1.140

0.00536

0.00632

0.00655

0.00868

0.00708

Total Dispensers: 04
0.005

0.00000

S20B

0.847

0.00386

S21A

1.289

0.00612

S21B

1.209

0.00571

S22A

1.803

0.00875

S22B

Total Dispensers: 08

0.846

0.00385

2nd Floor Swabs

Absorbance

S20A

3
2

STANDARD

2nd F (SC)

0
0

0.005

0.01

0.015

2nd F (SH)

Concentration (mg/mL)

Figure 3. Second Floor swabbing results graph

Table [Link] Floor Swabbing Results

Dispe
nsers
A

Absorb
ance
(AU)

(HOT)

Conce
ntratio
n

Dispe
nsers
B

(mg/ml)

(COLD)

Table [Link] Floor Swabbing Results

Absor
bance
(AU)

Conce
ntratio
n

Dispe
nsers
A

(mg/ml)

(HOT)

Absorb
ance
(AU)

Conce
ntratio
n

Dispe
nsers
B

(mg/ml)

(COLD)

Absor
bance
(AU)

Conce
ntratio
n
(mg/ml)

S03A

0.927

0.00427

S03B

1.360

0.00648

S05A

1.494

0.00717

S05B

2.065

0.01009

S04A

1.801

0.00874

S04B

1.632

0.00787

S06A

0.009

0.00000

S06B

1.716

0.00830

S09A

0.008

0.00000

S09B

0.984

0.00456

S12A

1.047

0.00488

S12B

0.498

0.00207

S10A

0.003

0.00000

S10B

1.432

0.00685

S13A

1.556

0.00748

S13B

0.473

0.00194

S17A

1.456

0.00697

S17B

1.535

0.00738

S19A

2.019

0.00985

S19B

1.723

0.00834

S18A

0.678

0.00299

S18B

1.840

0.00893

Total Dispensers: 05

Total Dispensers: 06

Absorbance

4th Floor Swabs

Absorbance

3rd Floor Swabs

3
2

STANDARD

3rd F (SC)

0
0

0.005

0.01

0.015

3
2

STANDARD

4th F (SC)

0
0

0.005

0.01

0.015

4th F (SH)

Concentration (mg/mL)

3rd F (SH)

Concentration (mg/mL)
Figure 4. Third Floor swabbing results graph

Figure 5. Fourth Floor swabbing results graph

Tables 2 to 5 contains data gathered from
swabbing method. Table 2 contains the data gathered
for the first floor swabs for both hot and cold nozzles.
Compared to the standard graph, the concentration of
the swab samples ranges from 0 mg/mL to 0.0087
mg/mL. Table 3 contains the data gathered for the
second floor swabs for both hot and cold nozzles.
Compared to the standard graph, the concentration of
the swab samples ranges from 0 mg/mL to 0.009
mg/mL. Table 4 contains the data for the third floor
swabs, which concentration ranges from 0 mg/mL to
0.00893 mg/mL. Lastly, Table 5 contains the gathered
data from the fourth floor, which ranges from 0 mg/mL
to 0.0101 mg/mL.

Table 6. First Floor Water Sampling Results

Dispe
nsers
A

Absor
bance
(AU)

Concen
tration
(mg/ml)

(HOT)

Dispe
nsers
B

Absor
bance
(AU)

Third Floor
Concen
tration

(COLD)

W01A

0.003

0.00000

W01B

0.417

0.00166

W07A

0.820

0.00372

W07B

0.399

0.00157

W20A

0.618

0.00269

W14B

0.306

0.00109

W15B

0.764

0.00343

W20B

0.583

0.00251

Second Floor
0.380

0.00147

0.475

0.00195

W03B

0.776

0.00349

W18A

0.356

0.00135

W04B

0.507

0.00212

W09B

0.460

0.00188

W10B

1.055

0.00492

W17B

0.274

0.00093

W18B

1.242

0.00588

(mg/ml)

First Floor

W16A

W04A

W11B

1.188

0.00560

W16B

0.328

0.00120

W23B

0.932

0.00429

Fourth Floor
W12A

0.006

0.00000

W05B

0.183

0.00046

W19A

0.934

0.00430

W06B

0.016

0.00000

W12B

1.019

0.00474

W19B

0.912

0.00419

3rd Floor Water Samples

2.5

2.5

1.5
STANDARD
1st F (WC)

Absorbance

Absorbance

1st Floor Water Samples

1.5
STANDARD
3rd F (WC)

1st F (WH)

3rd F (WH)

0.5

0.5

0.475
0.356

0
0

0.005

0.01

0.015

Concentration (mg/mL)

0.005

0.01

0.015

Concentration (mg/mL)

Figure 8. Third Floor water sampling results graph

2nd Floor Water Samples

4th Floor Water Samples

2.5

2.5

1.5
STANDARD
2nd F (WC)

Absorbance

Absorbance

Figure 6. First Floor water sampling results graph

1.5
STANDARD
1

4th F (WC)

0.934

2nd F (WH)
0.5

4th F (WH)
0.5

0.38

0
0

0.005

0.01

0.015

Concentration (mg/mL)

Figure 7. Second Floor water sampling results graph

0.006
0.005

0.01

0.015

Concentration (mg/mL)
Figure 9. Fourth Floor water sampling results
graph

Table 6 contains the unified data

gathered from first to fourth floor using water sampling
technique. It shows that the concentration from the
dispensers ranges from 0 mg/mL to 0.00372 mg/mL in
the first floor, 0.00120 mg/mL to 0.0056 mg/mL in
the second floor, 0.000092 mg/mL to 0.00588 mg/mL
in the third floor, and 0 mg/mL to 0.00474 mg/mL in
the fourth floor.
If we compare the data from both swabs and
water sample, we can see that swabbing samples have
a higher concentration than compared to those from
water sampling samples. But even after observing
that, we can see that the trend for both is almost the
same. The highest concentration that was obtained
was 0.0101 mg/mL from the dispenser from the fourth
floor, found at the west building.

Bacteriological quality of drinking water from

dispensers
In any case, the water that we drink also
comes with various types of contaminants in the form
of microbes and minerals. Among the common
minerals found in water are calcium and magnesium.
Fluoride is also added to the water to help prevent
disease and protect against tooth decay [8].
Researchers
at
Boston's
Northwestern
University tried the bacterial number in water coolers.
They found 2,000 potentially harmful organisms in
every thousandth liter of water. Its not that the water
wasn't fresh, water drawn from the bottles themselves
was significantly lower in bacteria, it is the fact that
the water dispensed through the cooler that was
largely infected. The reservoirs weren't clean, and
therefore, contaminated the water [9].

Twenty-three dispensers were monitored over a

period of two days to evaluate their hygienic status.
Two possible microorganisms present in dispensed
water are as follows:

1. Pseudomonas aeruginosa
This microorganism is a member of the
Gamma Proteobacteria class of Bacteria. It is a
Gram-negative, aerobic rod belonging to the
bacterial
family
Pseudomonadaceae.
Contamination from P. aeruginosa proved hard
to remove. Various studies have shown how
this microorganism in biofilm is significantly
less susceptible to the action of disinfectants
[10].
Pseudomonas
aeruginosa is
an
opportunistic pathogen, meaning that it
exploits some break in the host defenses to
initiate an infection. It causes urinary tract
infections,
respiratory
system
infections,
dermatitis, soft tissue infections, bacteremia,
bone and joint infections, gastrointestinal
infections and a variety of systemic infections,
particularly in patients with severe burns and
in cancer and AIDS patients who are
immunosuppressed. Pseudomonas aeruginosa
infection is a serious problem in patients
hospitalized with cancer, cystic fibrosis, and
burns. The case fatality rate in these patients
is near 50% [10].

Figure 11. Gram-stained P. aeruginosa bacteria [10]

Figure 10. Experimental Gram-stained Swabs

2. Escherichia coli
Escherichia coli is a Gram-negative,

anaerobic rod-shaped bacterium of

facultative

the

genus

Escherichia that is commonly found in the

lower intestine of warm-blooded organisms
like
humans. A growing body of researchers has
examined
E.
coli and
found
it
to
be
environmentally persistent microorganism which
can survive for extended periods outside of a host
[11].
Most E. coli are harmless and actually are
an important part of a healthy human intestinal
tract. However, some strains, such as E. coli O157:
H, can make people sick, causing severe stomach
cramps,
diarrhea
and
vomiting.
Serious
complications of an E. coli O157:H7 infection can
include kidney failure [12].

lowest absorbance is W01A (1st floor) with only 0.003

while the highest is W19A (4th floor) with 0.934
absorbance. In the cold water the one having the
lowest absorbance is W06B (4th floor) with only 0.016
while the highest is W18B (3rd floor) with 1.242
absorbance. In the data gather gathered we can say
that most clean among the sample W01A (1st floor)
when it comes to hot water while on cold water is
W06B (4th floor) we therefore conclude that the north
building is the cleanest among the south, west and
north buildings. Possible error in this experiment the
replacement of water containers which is done regular
intervals so the data we gather may vary on the time it
was collected.

References
[1][Link]
[2] [Link]
[3] [Link]
[4]
[Link]
[5][Link]
/common/downloads/[Link]
[6] University of Waterloo: Electrolytes. 1992-2015.
Figure 12. Gram-stained E. coli bacteria [12]

Conclusion
The Experiment was able to determine the
absorbance and concentration of microorganisms
through swabbing and water sampling. As stated from
the gathered data S20A (1st Floor) Swabbing result
absorbance is 0.005 which is the lowest while S19A
(4th Floor) absorbance is 2.019 which is the highest.
We can say that some of the errors in this experiment
is time, the time in this experiment is very essential
because the water dispenser is mostly used during
break time or after class. Another one is the
cleaning/disinfectant of the water dispenser by the
personnel which is only done twice a day or maybe
every other day or sometimes after a term which is we
do not know when they clean the water dispensers.
In the water sampling we categorized it by hot
or cold because we vary on the use of water depending
on what we need. In the hot water the one having the

[7] ChemWiki. (n.d.). Spectrophotometry. Retrieved

12
13,
2015,
from
[Link]
/Kinetics/Reaction_Rates/Experimental_Determination
_of_Kinetcs/Spectrophotometry
[8] [Link]
[9] [Link]
[10]
[Link]
[11] [Link]
[12] [Link]

Appendices
I.

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