Chapter 1

INTRODUCTION

Nitrates are naturally occurring. All rainfall and groundwater aquifers contain some

nitrate-nitrogen. However, contaminated rural water supplies provide a clue for discovering

other nitrate sources. Nitrate accumulates in agricultural watersheds where farmers spread

inorganic fertilizers and animal manures on cropland. Nitrogen not taken up by crops can leach

through the soil to groundwater and then flow to recharge areas or private wells. Residents in

rural communities typically use on-lot septic systems and some homeowners rely on lawn

fertilizers. These too can be sources of nitrate in drinking water. Health effects of nitrate in

drinking water are most significantly linked to methemoglobinemia, also known as blue-baby

syndrome. Nitrates in drinking water can also affect certain adults and small children. Pregnant

women can pass methemoglobin on to developing fetuses and low birth weights have been

attributed to high nitrates in water. However, nursing mothers do not pass nitrites to infants via

their milk. Children between the ages of 12 to 14 have shown delayed reactions to light and

sound stimuli from drinking water containing greater than 105 mg/l of nitrate. In general, studies

by the World Health Organization and the National Academy of Sciences reveal that

consumption of nitrates in drinking water does not represent a significant health risk to the adult

population. Nitrates level in drinking water is recommended below 50 mg/L, especially for

babies. Indeed, their digestive systems are immature, and thus more likely to allow the reduction

of nitrate to nitrite. The nitrite in the digestive tract of babies can cause methenoglobinemia.

Nitrates can be removed by ion exchange using strong anionic resin in Cl- form. Reverse

Osmosis is also efficient to remova nitrates, although the hydrophilic properties of the molecule

1
does not allow an optimal removal. Biological denitrification is used in municipal waste water

treatment plant.

Once a water supply becomes contaminated with nitrate, it is costly to treat. While

treatment to meet drinking water needs is practical, treatment of larger quantities like livestock

supplies is costly. Ion exchange units, reverse osmosis, or distillation all remove nitrate from

drinking water. Note that boiling water does not remove nitrates and is not a treatment

alternative. In fact, it increases nitrate concentrations as water evaporates.

An ion exchange unit operates much like a household water softener. A softener filters

calcium and magnesium laden water through a resin coated with sodium ions. As water flows

through the unit, the resin releases its sodium ions and readily trades them for the calcium and

magnesium. For nitrate removal, the resin exchanges chloride ions for nitrate and sulfate ions in

the water. After treating many gallons of water, the resin will run out of chloride. Regenerating

the resin with a concentrated solution of sodium chloride (you can use bicarbonate instead of

chloride) recharges it for further treatment.

Ion exchange does have drawbacks. Because the resin prefers to absorb sulfate, water

high in sulfates hinders the nitrate exchange and reduces system effectiveness. If the resin

becomes saturated, it releases the nitrates in place of sulfates, resulting in an increased nitrate

concentration in the treated water. Also, nitrate ion exchange can make the water corrosive.

Neutralizing the water after it leaves the unit reduces this effect. Finally, ion exchange can be

expensive and requires maintenance. Since the backwash brine will be high in nitrates, care must

be given to its disposal. Reverse osmosis is another treatment mechanism. As water enters the

unit under pressure, it pushes against a cellophane-like plastic sheet or cellulosealso called a

2
semipermeable membrane. The membrane acts like a sieve, leaving ions like nitrates on one side

and allowing ion-free water to pass through the membrane. How well the membrane filters the

water is measured by the rejection rate. That is, how much nitrate will the membrane reject?

Estimates are around 83 to 92 percent of the incoming nitrate. Naturally, knowing the original

nitrate concentration in the water is important. If nitrate-nitrogen levels are extremely high

(greater than 110 mg/l) up to 90 percent may be removed. Unfortunately, remaining nitrate-

nitrogen may still result in drinking water that exceeds the standards of 10 mg/l nitrate-nitrogen.

Chapter 2

DESIGN AND METHODOLOGY

The method employed in the determination of the nitrate content of water sample is the

Ultraviolet Spectrophotometric Screening method.

Principle:

UV absorption method has the advantage of being simple, inexpensive, and chemical-

free. The measurement is a direct spectrophotometric determination of nitrate concentration,

where the magnitude of the absorbance at a wavelength of 220 nm is proportional to the

concentration of nitrate ions in solution, however, the measurement is subject to interferences

from inorganic and organic substances that reduce the transmission of light at wavelengths

similar to nitrate, including nitrite, bromide, chromophores dissolved organic carbon (DOC), and

turbidity. Because of this, a second measurement made at 275 nm may be used to correct the

NO3 value since NO3 does not absorb at 275 nm.

3
 Sample filtration is intended to remove possible interference from suspended particles.

Acidification with 1N HCl is designed to prevent interference from hydroxide or carbonate

concentrations up to 1000 mg CaCO3/L. Chloride has no effect on the determination.

Reagents:

a.) Nitrate-free water: redistilled or distilled, deionized water of highest purity to prepare all

solutions and dilutions.

b.) Stock nitrate solution: Potassium nitrate (KNO3) was dried in an oven at 105C for 24

hours. Then, 0.7218 g of it was dissolved in water and diluted to 1000 mL; 1.00 mL =

100 g NO3-N.
c.) Intermediate nitrate solution: 100 mL stock nitrate solution was diluted to 1000 mL with

water; 1.00 mL = 10.0 g NO3-N.

d.) Hydrochloric acid solution, HCl, 1 N.

Procedure:

1 mL of 1N HCL was added to a 50 mL clear sample then it was mixed thoroughly.

Afterwards, the absorbance at 220 nm and 275 nm was read and recorded. This procedure was

repeated three times.

Standard curve:

NO3 calibration standards were prepared in the range of 0 to 7 mg NO 3-N/L by diluting to

50 mL the following volumes of intermediate nitrate solution: 0, 1.00, 2.00, 4.00, 7.00 35.0

mL. The NO3 standards were treated in the same manner as samples.

Calculation:

For samples and standards, the absorbance due to NO 3 was obtained by subtracting the

absorbance reading at 275 nm two times from the absorbance reading at 220 nm. A standard

curve was constructed by plotting the absorbance due to NO 3 against NO3-N concentration of

standard. Calibration standard that did not follow the trend was disregarded. For standards and

4
samples whose correction value exceeds 10%, the absorbance reading at 220 nm was used

instead of the computed actual absorbance. The sample concentrations were obtained directly

using the equation of the standard curve.

Chapter 3

RESULTS AND DISCUSSION

Table 3.1 Determination of Nitrates Concentration in ppm for Wednesday Class

Group
Sampling A A Actual Concn
Numbe Trial A %E
Point 220nm 275nm A ppm
r
1 0.527 0.027 0.473 10.247 0.527 21.8619
2 0.477 0.026 0.425 10.901 0.477 19.7699
ORCHID
1 3 0.513 0.031 0.451 12.086 0.513 21.2762
STREET
4 0.494 0.022 0.45 8.907 0.45 18.6402
AVERAGE 20.3870

1 1.234 0.036 1.162 5.835 1.162 43.4310

ASIN 2 1.334 0.031 1.272 4.648 1.272 53.0335
BRIDGE 2 3 1.334 0.036 1.262 5.397 1.272 52.6151
#2 4 1.239 0.027 1.185 4.358 1.185 49.3933
AVERAGE 50.8682

1 0.958 0.031 0.896 6.472 0.896 37.3013

ASIN 2 1.123 0.031 1.061 5.521 1.061 44.2050
BRIDGE 3 3 1.138 0.039 1.06 6.854 1.06 44.1632
#1 4 1.118 0.034 1.05 6.082 1.05 43.7448
AVERAGE 42.3536

1 0.310 0.025 0.26 16.129 0.31 12.7824

2 0.368 0.035 0.298 19.022 0.368 15.2092
DACAG
4 3 0.318 0.028 0.262 17.610 0.318 13.1172
BRIDGE
4 0.318 0.034 0.25 21.384 0.318 13.1172
AVERAGE 13.5565

CHESKA 5 1 0.369 0.044 0.281 23.848 0.369 15.5210

STORE 2 0.410 0.047 0.316 22.927 0.410 16.9665
3 0.369 0.042 0.285 22.764 0.369 15.2510

5
Sampli Group
A A Actual Concn
ng Numbe Trial A %E
220nm 275nm A ppm
Point r
1 0.077 0.026 0.025 67.532 0.077 3.0335
AGUYAD 2 0.061 0.023 0.015 75.410 0.061 2.3640
1
BRIDGE 43 0.373
0.066 0.042
0.023 0.289
0.02 22.520
69.697 0.373
0.066 15.4184
2.5732
AVERAGE 15.7218
2.6569

PACAC 6 11 0.716
0.212 0.043
0.030 0.63
0.152 12.011
28.302 0.716
0.212 29.7699
8.6820
IRISAN
BRIDGE 22 0.623
0.205 0.041
0.027 0.541
0.151 13.162
26.341 0.623
0.205 25.8787
8.3890
2
BRIDGE 33 0.317
0.214 0.042
0.024 0.233
0.166 26.498
22.430 0.317
0.214 13.0753
8.7657
4 0.378 0.043 0.292 22.751 0.378
AVERAGE 15.6276
8.6122
AVERAGE 21.0879
1 0.216 0.032 0.152 29.630 0.216 8.8494
LASONG 2 0.223 0.037 0.149 33.184 0.223 9.1423
3
BRIDGE 3 0.228 0.031 0.166 27.193 0.228 9.3515
AVERAGE 9.1144

1 0.312 0.135 0.042 86.538 0.312 12.8661

BUCAN 2 0.315 0.132 0.051 83.810 0.315 12.9916
4
BRIDGE 3 0.293 0.139 0.015 94.881 0.293 12.0711
AVERAGE 12.6429

1 0.220 0.037 0.146 33.636 0.220 9.0167

PALM 2 0.222 0.036 0.150 32.432 0.222 9.1004
5
BRIDGE 3 0.213 0.033 0.147 30.986 0.213 8.7238
AVERAGE 8.9467

1 0.084 0.017 0.050 40.476 0.084 3.3264

NEVER-
2 0.086 0.019 0.048 44.186 0.086 3.4100
LAND 6
3 0.087 0.020 0.047 45.977 0.087 3.4519
RESORT AVERAGE 3.3961
Sampling Group A A Actual Concn
Trial A %E
Point Number 220nm 275nm A ppm
1 0.077 0.026 0.025 67.532 0.077 3.0335
AGUYAD 2 0.061 0.023 0.015 75.410 0.061 2.3640
1
BRIDGE 3 0.066 0.023 0.02 69.697 0.066 2.5732
AVERAGE 2.6569

1 0.212 0.030 0.152 28.302 0.212 8.6820

IRISAN 2 0.205 0.027 0.151 26.341 0.205 8.3890
2
BRIDGE 3 0.214 0.024 0.166 22.430 0.214 8.7657
AVERAGE 8.6122

1 0.216 0.032 0.152 29.630 0.216 8.8494

LASONG 2 0.223 0.037 0.149 33.184 0.223 9.1423
3
BRIDGE 3 0.228 0.031 0.166 27.193 0.228 9.3515
AVERAGE 9.1144

1 0.312 0.135 0.042 86.538 0.312 12.8661

BUCAN 2 0.315 0.132 0.051 83.810 0.315 12.9916
4
BRIDGE 3 0.293 0.139 0.015 94.881 0.293 12.0711
AVERAGE 12.6429 6

1 0.220 0.037 0.146 33.636 0.220 9.0167

PALM 2 0.222 0.036 0.150 32.432 0.222 9.1004
5
BRIDGE 3 0.213 0.033 0.147 30.986 0.213 8.7238
Table 3.2 Determination of Nitrates Concentration in ppm for Friday Class

7
Sampling Group A A Actual Concn
Trial A %E
Point Number 220nm 275nm A ppm
1 0.344 0.015 0.314 8.721 0.314 12.9498
ATAD
2 0.342 0.017 0.308 9.942 0.308 12.6987
HANGING 1
3 0.363 0.014 0.335 7.713 0.335 13.8285
BRIDGE AVERAGE 13.1590

19.42
1 0.175 0.017 0.141 0.175 7.1339
9
ASIN 22.36
2 0.152 0.017 0.118 0.152 6.1715
HOT 2 8
SPRINNG 19.04
3 0.147 0.014 0.119 0.147 5.9623
8
AVERAGE 6.4226

29.09
1 0.055 0.008 0.039 0.055 2.1130
1
LUMIC- 31.03
2 0.058 0.009 0.040 0.058 2.2385
NENG 3 4
BRIDGE 41.86
3 0.043 0.009 0.025 0.043 1.6109
0
AVERAGE 1.9875

14.72
1 0.258 0.019 0.220 0.258 10.6067
9
COPLO 15.20
4 2 0.263 0.020 0.223 0.263 10.8159
BRIDGE 9
3 0.292 0.017 0.258 11.644 0.292 12.0292
AVERAGE 11.1506

1 0.336 0.011 0.314 6.548 0.314 12.9498

RIVER 2 0.331 0.005 0.321 3.021 0.321 13.2427
5
VIEW 3 0.334 0.010 0.314 5.988 0.314 12.9498
AVERAGE 13.0474

24.81
1 0.266 0.033 0.200 0.266 10.9414
2
27.27
UP- 2 0.264 0.036 0.192 0.264 10.8577
6 3
STREAM
26.81
3 0.276 0.037 0.202 0.276 11.3598
2
AVERAGE 11.0530
Table 3.3 Determination of Nitrates concentration in ppm for Saturday Class

8
 The concentration of nitrate is dependent on the actual absorbance indicating that as the

actual absorbance increases the concentration increases. High concentrations of nitrates in

Wednesday class and low concentrations in Friday class while a combination in Saturday class.

The Asin Bridge has the highest concentration of nitrates. Sampling points of the higher

concentrations flows on the same route which means that the source of water flowing on that

area uses high concentration of nitrates. Usually this value is cause by the agriculture

plant or leaking in the seawage system. Therefore, this indicate that the

sample is come from the area where there are population and plantation

exists. The lowest nitrate content is the Lumicneng Bridge which can be

considered as least not polluted among all as its average is 1.9875ppm. Ten

out of the 18 sample points are polluted as their concentrations are higher to

the standard value of 10ppm nitrates in water. From Orchid Bridge to Pacac Birdge

gave a high result of concentration of nitrates with an average of 27.33 ppm. The Aguyad Birdge

to Neverland resort gave a value of 9.47 ppm in average and Atad Hanging Birdge to Up-Stream

of 7.49 ppm.

There are few factors that contributes to the increasing in the values of

nitrate in the water. The common sources are from the agricultural runoff,

refuse dump runoff or contamination with human or animal wastes. The

concentration often fluctuates with the season and may increase when the

river is fed by nitrate-rich aquifers.

However, if the concentration of the nitrate in the water exceeded the

requirement, the water is dangerous to be consumed by the human body.

Meanwhile, excess nitrate and nitrite in water also cause depletion of oxygen

9
in the water. This will cause the aquatic habitats to die and at the mean time

effects the ecosystem.

Chapter 4

CONCLUSION AND RECOMMENDATION

In conclusion, the values obtained varies from each sampling point. Ten samples have greater

amount exceeding the maximum allowable amount for nitrated which is 10mg/L. The result

showed the Asin Bridge has the highest concentration of nitrates whereas the Lumicneng Bridge

has the least. From Orchid Bridge to Pacac Birdge gave a high result of concentration of nitrates

with an average of 27.33 ppm. It could be caused by the domestic wastes such as agricultural

activities where inorganic fertilizers and animal manures are used, use a lot septic systems, and

human wastes dumped in the water system and therefore polluted. The Aguyad Birdge to

Neverland resort gave a value of 9.47 ppm in average and Atad Hanging Birdge to Up-Stream of

7.49 ppm.

Nitrate in drinking water can be a problem, especially for infants. A water test is the only

way to determine whether the nitrate-nitrogen concentration is under the acceptable standard of

10 mg/L. Proper well location and construction are key practices to avoiding nitrate

contamination of drinking water. Management practices to reduce the risk of contamination from

applied fertilizers help keep the water supply safe.

If drinking water exceeds the acceptable nitrate-nitrogen standard, the choices are to use

an alternate water supply or treat the water. An alternate supply may be bottled water for

drinking, especially for infant formula, or a new well in a different location or aquifer. Water

10
treatment options are distillation, reverse osmosis, or ion exchange.

When the sample were stored for sometimes, it is highly recommended to shake first the

sample on its container before performing the experiment to achieve precise and accurate results.

When measuring absorbance in spectrophotometer it should 0 absorbance for the blank solution

as a reference, before putting sample in the cuvette.

REFERENCES

Standard Methods for the Examination of Water and Wastewater; APHA, AWWA, and

st
WEF, 21 Edition, 2005.
Methods for the Examinations of Waters and Associated Materials: General Principles of

Sampling and Accuracy of Results. 1980. Her Majestys Stationery Off., London,

England.
Jensen, V.B., Darby, J.L., Seidel, C. & Gorman, C. (2012) Drinking Water Treatment for

Nitrate. Technical Report 6 in: Addressing Nitrate in Californias Drinking Water with a

Focus on Tulare Lake Basin and Salinas Valley Groundwater. Report for the State Water

Resources Control Board Report to the Legislature. Center for Watershed Sciences,

University of California, Davis.

Sahli, M.A.M., Annouar, S., Mountadar, M., Soufiane, A. & Elmidaoui, A. (2008) Nitrate

removal of brackish underground water by chemical adsorption and by electrodialysis.

Desalination, 227, 327333.

11
 Bilidt, H. (1985) The use of reverse osmosis for removal of nitrate in drinking water.

Desalination, 53, 225230.

APPENDICES

APPENDIX A

List and Uses of Apparatus

Spectrophotometer
- It is used at 275 nm with matched silica cells of 1-cm or longer light path, for the

reading of the absorbance against a blank reference (distilled water) at 0 absorbance

Graduated Cylinder (50mL and 10 mL)
- It was used to accurately measure the liquids used.
Graduated Pipette
- It was used to transport a measured amount of volume of liquid to the Erlenmeyer

flasks.
Pippetol
- It was was used to suck the liquids through the pippete.
Erlenmeyer Flask
- It was used as a container for the different solutions and for mixing the solutions

made.

APPENDIX B

Definition of Terms

12
 Physical properties of water quality include temperature and turbidity. Chemical

characteristics involve parameters such as pH and dissolved oxygen. Biological

indicators of water quality include algae and phytoplankton.

Nitrate ion (NO3) is the stable form of combined nitrogen for

oxygenated systems. Although chemically unreactive, it can be

reduced by microbial action.

Nitrite ion (NO2) contains nitrogen in a relatively unstable oxidation

state.
Conductivity (or specific conductance) of an electrolyte solution is a measure of its

ability to conduct electricity. The SI unit of conductivity is siemens per meter (S/m).
pH is a determined value based on a defined scale, similar to temperature. This means

that pH of water is not a physical parameter that can be measured as a concentration or in

a quantity. Instead, it is a figure between 0 and 14 defining how acidic or basic a body of

water is along a logarithmic scale.

APPENDIX C

Computations

Asin Hot spring bridge Absorbance at 220 nm Absorbance at 275 nm

Trial 1 0.292 0.019
Trial 2 0.263 0.020
Trial 3 0.258 0.017
Table C.1 Data for Coplo bridge

Computations for Absorbance (A)

Formula: A = [Absorbance at 220nm 2(absorbance at 275 nm)]

Trial 1: A = 0.292 2(0.019)

A = 0.254
Trial 2: A = 0.263 2(0.020)

13
 A = 0.223
Trial 3:
A = 0.258 2(0.017)
A = 0.224
Computations for E %

Formula: E% = [ Absorbance at 220 ]

( Absorbance at 220 A )
x 100

Trial 1:

E% = [ ( 0.292 0.254 )
0.292 ]
x 100

E% = 13. 0137
Therefore, Actual Absorbance =0.292

Trial 2:

E% = [ ( 0.263 0.223 )
0.263 ]
x 100

E% = 15. 2091
Therefore, Actual Absorbance = 0.263

Trial 3:

E% = [ ( 0.258 0.224 )
0.258 ]
x 100

E% = 13.1783 Therefore Actual Absorbance = 0.224

Computations for ppm

[ A 0.0045]
Formula: A = 0.0239(X) + 0.0045; X = [0.0239]

14
 Where X = Concentration of nitrates in ppm
A = Actual Absorbance

Trial 1:
[0.292 0.0045]
X= [0.0239]

X = 12.0239 ppm

Trial 2:
[0.263 0.0045]
X= [0.0239]
X = 10.9854 ppm

Trial 3:
[0.258 0.0045]
X= [0.0239]
X = 10.7782 ppm

12.0239+10.9854 +10.7782
Average: 3

= 11.2625 ppm

Figure C.1 Calibration Curve of Coplo Bridge

15
16
 APPENDIX E

Problems

1. What per cent of the incident light at a given wavelength is transmitted by a medium

which at a given wavelength has an absorbance of 1.176?

Given:
A= 1.176
Required: %T
Solution:
100
A = - log ( %T

100
1.176 = -log ( %T )

%T = 1499.6848 %

2. What is the absorbance of a solution which absorbs 2/3 of the incident light?

Given:
Required: A
Solution:

2
A =- log ( 3

A=

3. A 7.5x 10
5
F KMnO 4 has an absorbance of 0.658 when measured in a 1.5 cm

cell at 525 nm. (a) Calculate the molar absorptivity, , of KMnO 4 on the basis of these

data. (b) Calculate the absorptivity, a, at this wavelength when the concentration is

17
expressed as ppm KMnO 4 . (c) Calculate the %T of the above solution if the

measurement had been made in a 1.00 cm cell.

Given:
c = 7.5x 10
5
F KMnO 4
A = 0.658
b = 1.5 cm
Required:

a.
b. a
c. %T
Solution:

a. A = bc

A
= bc

0.658
mol
=
(
( 1.5 cm ) 7.5 x 105
L )
= 5848.8889

L
b. A = abc
molcm
A
a = bc
0.658
mol
158.034 g
a= L
1 L
5 1 mol
(1.5 cm)(7.5 x 10 )
1000 mL

a = 37010.31986

18
L
gcm
 100
c. A = - log ( %T

L 5 mol
A = 5848.8889 molcm *( 7.5 x 10 L (1.0 cm)

A = 0.4387

%T = 274.5997

19