TITLE: DETERMINATION OF AMOUNT OF NITRATES

AT LOVINGTON STREAM IN KIPSONGO, KITALE

NAME: CHEMWILE A ONESIMUS

COURSE: DIPLOMA IN ANALYTICAL CHEMISTRY

CENTER CODE: 503111

COURSE CODE: 2915

PAPER CODE: 307/3

INDEX NUMBER: 5031112311

INSTITUTION: KITALE NATIONAL POLYTECHNIC

PRESENTED TO: KENYA NATIONAL EXAMINATION

COUNCIL IN PARTIAL FULFILLMENT FOR THE
AWARD OF DIPLOMA IN ANALYTICAL CHEMISTRY
SUPERVISOR: MR WALLACE KARANJA

SERIES: NOVEMBER

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 DECLARATION
This research project is my original work, all the authors whose work has been referred to have
been properly cited and the references provided. This work has not been presented elsewhere for
award of a diploma or any other awards
NAME: CHEMWILE A ONESIMUS
Signature: _______________________
Date: ___________________________

SUPERVISOR APPROVAL
This project has been submitted for examination with my approval us a technical supervisor
Signature: _______________________
Date: ___________________________
Name: __________________________
Department of Applied Science,
Kitale National Polytechnic

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 ACKNOWLEDGEMENT
First I would like to thank the almighty Father God for the much He has done for me throughout
my study period, secondly I would like to thank my supervisor Mr. Wallece Karanja and
technicians classmates for the guidance and support they gave me in writing this research
project, I would like to thank my colleagues who assisted me in discussions and hence made my
research successful, Finally but not least I want to thank my parents Mr. Fred Chemwile and
madam Esther Murunga for the support they offered, they took care of my financial requirements
during the research of this project and gave me enough advice on my academic journey
Many other people contributed valuable help and support though impossible to cite them all but
they shall always be cherished and remembered in my heart.

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Table of Contents
DECLARATION..............................................................................................................................................2
ACKNOWLEDGEMENT.................................................................................................................................3
ABSTRACT....................................................................................................................................................4
CHAPTER ONE..............................................................................................................................................5
1.0 INTRODUCTION.................................................................................................................................5
1.1 Background information....................................................................................................................5
1.1 Statement of the problem.................................................................................................................6
1.1.1 Objectives...................................................................................................................................6
1.1.2 General objectives......................................................................................................................6
1.1.3 Specific objectives......................................................................................................................6
1.2 Justification and significance of the study.........................................................................................6
CHAPTER TWO.............................................................................................................................................7
2.0 Literature review...............................................................................................................................7
2.1.1 Nitrates pollution effects.............................................................................................................7
2.2. Industrial removal of Nitrates in wastewater...................................................................................8
2.2.1. Physical and chemical method...................................................................................................8
2.2.2 Chemical method........................................................................................................................9
2.2.3 Bio-chemical method..................................................................................................................9
CHAPTER THREE........................................................................................................................................10
3.0 Methodology and materials.............................................................................................................10
3.1 Sampling site...................................................................................................................................10
3.2 Material and apparatus...................................................................................................................10
3.2.1 Glassware.................................................................................................................................10
3.2.2 Sample containers.....................................................................................................................10
3.3 Sampling..........................................................................................................................................10
3.4 Experimental....................................................................................................................................10
3.4.1 Reagents and preparations........................................................................................................10
3.4.2 Reagents preparation.................................................................................................................11
3.4.3 Stock and working solution preparation....................................................................................11
3.4.4 Procedure for Nitrate analysis...................................................................................................11
CHAPTER FOUR..........................................................................................................................................12
4.0 Results and discussion of results.....................................................................................................12

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 4.1 Graph of highest absorbance for Nitrates.......................................................................................12
4.1.1 Graph for Nitrates standards.....................................................................................................13
4.1.2 Table for concentration of Nitrates in the stream samples........................................................13
4.2 Discussions......................................................................................................................................13
4.3 Discussion of beer Lambert’s law and it’s application in the research.............................................13
4.4 Basic instrumentation of a Spectrophotometer..............................................................................14
4.5 Application of Beer Lambert’s law...................................................................................................14
CHAPTER FIVE............................................................................................................................................15
5.0 Conclusion and Recommendations..................................................................................................15
5.1 Conclusions......................................................................................................................................15
5.2 Recommendations...........................................................................................................................15
LIST OF REFERENCES..................................................................................................................................16

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 ABSTRACT
Kipsongo, Kitale falls under Mediterranean climate, its major economic activities are farming
and pastoralism.
Introduction of irrigation farming in the region with use of synthetic fertilizers has significantly
increased levels of Nitrates in water streams in this region, which is due to the fertilizers runoff
from the agricultural areas, water pollution by Nitrates could be a major problem in this region,
however the level of this possible pollution has never been evaluated, the main objective of this
research is to evaluate how change in land use in Kipsongo Kitale affects the quality of stream
water.
Sampling was done in the following four water points Kipsongo river, river athi, kimana swamp
and Lovington stream, presence of Nitrates were determined spectrophotometric ally at
wavelength of 420nm using Uv/vis Spectrophotometer, the analysis was based on the reaction of
Nitrates with sodium salicylate in sulfuric acid medium, sodium hydroxide tartrate solution was
added forming yellow colored salts nitro salicylic acid
Nitrates concentration ranged from 3.53_+ 0.11mg/l to 6.34_+ 0.16mg/l kimana swamp,
5.32_+0.04mg/l to 6.30_+0.26mg/l, Lovington stream 3.72_+ 0.17mg/l to 4.02_+ 0.40mg/l and
river athi 4.30_+0.15mg/l to 9.0_+ 0.20mg/l
The results show that concentrations of Nitrates is below 50mg/l as recommended by WHO, the
study concludes that Nitrates in stream water of Kipsongo Kitale pause health hazards to
residents who use water for drinking and cooking, in view of this, it is recommended that the
health ministry need to raise level of awareness on the health hazards paused by water polluted
with Nitrates in the region.

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 CHAPTER ONE
1.0 INTRODUCTION
1.1 Background information
Stream water has been used for domestic work for long time.The current increase of human
civilization has put serious concern to the safety of stream water for drinking purpose, this is
mainly because of the current rate of pollution.
This form of environmental degradation occurs when pollutants are directly or indirectly
discharged into stream without treatment to remove harmful compounds. Water pollution is a
major global problem mainly because of its serious effects on the entire biosphere.
The specific contaminates leading to pollution of water include a wide range of chemicals,
pathogens or physical changes such as elevated temperature and discoloration.
High concentration of naturally occurring substances can have negative impact on aquatic life.
Currently the high water pollution load comes from chemicals which include detergents,
disinfectants, volatile organic compounds, chlorinated solvents, perchlorate pharmaceutical
drugs and their metabolites, ammonia from food processing waste, chemical waste such as
industrial by-products and fertilizers.
Nitrates (NO3) are among the major inorganic pollutants contributed by nitrogenous fertilizers,
organic manures, human and animal wastes and industrial effluents through the biochemical
activities of microorganisms.
Excessive use of nitrogenous fertilizers such as Calcium Ammonium Nitrate (CAN) in
agriculture has been of the primary source of high nitrate ions on water source, Apart from
nitrate ions, nitrogen is applied in ammonium (NH+4) and amide (NH-2) forms, which generate
nitrate in soil system

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1.1 Statement of the problem
Lovington stream in Kipsongo could be polluted by nitrates from untreated sewage and animal
waste, however the introduction of agricultural activities in the area has not been properly
regulated leading to very high levels of river pollution by these pollutants, this result from use of
synthetic fertilizers in irrigation schemes
This research aims to raise level of awareness in regards to use of synthetic fertilizers in
irrigation schemes
1.1.1 Objectives
 The following objectives will be discussed.
1.1.2 General objectives
 To evaluate how change in land use in KIPSONGO area affects the quality of stream
water in Kitale
1.1.3 Specific objectives
 To determine the level of nitrates in stream water in Kipsongo
 To establish if the level of nitrates in stream water in Kipsongo Pause any health hazards
to the residents who use them for drinking and cooking

1.2 Justification and significance of the study

There have been reports of nitrate pollution across the world many lakes and rivers are at threat
of vanishing as a result of eutrophication, there have been insufficient researches in the world on
these contaminates with little solutions offered
Stream water in Kipsongo are similarly polluted by nitrates caused by a number of sources
including agricultural activities, industrial effluents, and untreated sewage and animals wastes.
There is need for intensive research into the long term implication of these water pollutants to
environment, animals and humans

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 CHAPTER TWO
2.0 Literature review
Nitrate and nitrite are naturally occurring ions that are part of the nitrogen cycle. The nitrate ion
(NO-3) is the stable form of combined nitrogen for oxygenated system, although chemically
nonreactive it can be reduced by microbial action.
The nitrite ion (NO-2) contains nitrogen in a relatively unstable oxidation state, chemical and
biological processes can further reduce nitrite to various compounds or oxidize it to nitrate.
Nitrate ion has molecular formula mass of 62.0049g/mol, Nitrates also describe the organic
functional group RONO2. Nitrate are mainly produced for use as fertilizers in agriculture because
of their high solubility and biodegradability. The main Nitrates are ammonium, sodium,
potassium and calcium salts. Almost all inorganic Nitrate salts are soluble in water at standard
temperature and pressure
A rich source of inorganic Nitrate in the human body comes from diets rich in leafy green foods,
such as spinach and arugula.
Agriculture activities, primarily crop and livestock production, account for over 80% of all
nitrogen added to the environment.
Agricultural areas have the highest rates of Nitrate contaminated
Animal manure is the second largest source of Nitrates in the environment, non-agricultural
sources of nitrogen contribute less than 20% of the nitrogen released into the environment of
this, 6% is released from point sources (basically pipes) into water bodies while 14% is deposited
from atmospheric sources. Point sources in urban watersheds can cause significant localized
Nitrate problems or individual wells, municipal sewage plants account for 80% of point source
Nitrates discharges, individual septic tanks and a number of industrial source account for the rest.
2.1.1 Nitrates pollution effects
Nitrates are absorbed from the small intestine into the body, there is little absorption from the
stomach in most species, although some has been reported from the human of cattle. If
concentration of Nitrates exceed 50mg/l. Drinking water can cause methemoglobinemia and
have been linked to gastric and intestinal cancer, Methemoglobinemia most often affects infant
of less than six months of age, Methemoglobinemia is the condition in the blood which causes
infant blue baby syndrome met hemoglobin is probably formed in the intestinal tract of an infant
when a bacterium converts the Nitrates ion to nitrite ion , One nitrite molecule then reacts with
two molecules of hemoglobin to form met hemoglobin , This altered form of blood protein
prevents the blood cells from absorbing oxygen which leads to slow suffocation of the infant
which may lead to death rapidly because of the oxygen deprivation, the infant will often take on
a blue tinge in the lips and extremities hence the name blue baby syndrome, Other signs of
methemoglobinemia are gastrointestinal disturbance such as vomiting and diarrhea, relative
absence of distress when severely cyanotic but irritable when mildly cyanotic and chocolate
brown colored blood

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Nitrate pollution has also been associated with stomach and gastrointestinal cancer, Although
many studies have been performed attempting to link stomach and gastrointestinal cancer to
Nitrate intake, There is no conclusive evidence for correlation researchers claim that Nitrate
represents such a potential risk because of nitration reaction which with appropriate substrates
present, From N-nitrous compounds which are strongly carcinogen in animals. Nitrate pollution
also affects the environment negatively, the widely known environmental adverse effect of
Nitrate pollution is eutrophication, This is the process by which a body of water acquires a high
concentration of nutrients, especially Nitrates, these typically promote excessive growth of algae,
As the algae die and decompose, high levels of organic matter and the decomposing organisms
deplete the water of available oxygen causing the death of other organisms such as fish.
Another environmental effect is anoxia, Anoxia is lack of oxygen caused by excessive nutrients
in waterways which triggers algae growth, when the plants die and decay, oxygen is stripped
from the water, which then turns green or milky white and gives off a strong rotten egg odor, the
lack of oxygen is often deadly for invertebrates, fish and shellfish.
Due to these health risks a great deal of empty sis has been placed on finding effective treatment
process to reduce Nitrate concentrations to safe levels, An even more important facet to reduce
the problem is prevention measures to stop the leaching of Nitrate from the soil some suggest
that reducing the amount of fertilizers used in agriculture will help alleviate the problem and may
not hut crop yields, Other new developments in leach pits and slurry stores help to control the
nitrate that comes from stored manure, by installing these prevention methods and reducing the
amount of fertilizer used the concentration of Nitrates in the ground water can be reduced over
time , treatment processes such as ion exchange can have an immediate effect on reducing levels
in drinking water, These processes do not remove the entire Nitrate but can help to bring the
concentration down to the suggested level of 10mg/l
Structure of nitrate ion

2.2. Industrial removal of Nitrates in wastewater

Untreated wastewater quite often finds its way into rivers and streams, having its major source
from domestic water usage and industrial effluents, wastewater carries a lot of Nitrates many
researchers have been done to develop effective method for removal of these water pollutants.

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2.2.1. Physical and chemical method
2.2.1.1 Removal of nutrients from wastewater using polymer hydro gels
Besides the primary and secondary treatment the removal of nutrients from the wastewater can
also be achieved by applying the inorganic and polymeric sorbents as the absorbents of nutrients
in water, However the conventional absorbents such as clay minerals, zirconia, titanium
polymeric ligand exchange and activated alumina are not feasible in practical wastewater
treatment due to their insufficient absorption capacities and not fully developed usage procedure.
2.2.2 Chemical method
2.2.2.1 Electrocoagulation method
Electrocoagulation is a wastewater treatment technique which involves the application of electric
current to the sacrificial electrodes, mostly iron and aluminum, resulting in coagulation.
2.2.2.2 Chemical precipitation
Chemical precipitation is one of the most widely used methods to remove Nitrates by adding
coagulant into wastewater, the most often employed multivalent metal ions are the compounds of
calcium, Aluminum and iron
2.2.3 Bio-chemical method
The Nitrate can be removed from water or wastewater by physio-chemical and biological
treatment methods, The conventional treatment processes which are mostly applied in removal of
Nitrate include reverse osmosis, ion exchange, electro dialysis and activated carbon absorption in
addition there is the metallic iron-aided abiotic Nitrate reduction which has been reported.
The biological denitrification method gives better removal of Nitrate, Which involve the
respiratory process of facultative anaerobes without oxygen in this method the denitrifying
bacterial reduce the inorganic compounds ( Nitrate and Nitrite) into harmless nitrogen gas ,
Therefore no further treatment is needed to get rid of the waste moreover the denitrifying
bacteria are ubiquitous in nature this enables the microbial removal of Nitrate from the
wastewater to be operated easily at low cost , Although the biological denitrification is the most
environmental friendly and cost effective method, It usually needs longer time for the removal of
highly concentrated nitrate.

CHAPTER THREE
3.0 Methodology and materials
3.1 Sampling site
 Kipsongo river

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  Lovington stream
 Kimana swamp
 River athi
3.2 Material and apparatus
3.2.1 Glassware
For the best results all glassware involved in the test will be washed with 0.1M HCl thoroughly
rinsed with distilled water, This was done to knock off impurities that may adhere to glass and
bottles ,
At each sampling site the bottles will be rinsed three times with the stream water before taking
samples
3.2.2 Sample containers
Plastic containers with high density polyethylene will be used as they withstand breakage and
less impurities adherence
3.3 Sampling
A sample of about 150ml will be collected at each sampling point, the volume of samples will be
small to reduce cost of transport and impurities
The water samples were collected from below, middle and at the shore of the stream
The samples will be kept for in refrigerator for the period spend in sampling
3.4 Experimental
3.4.1 Reagents and preparations
3.4.1.1 Reagents for Nitrates test
 Sodium tartrate
 Sodium hydroxide
 Sodium salicylate
 Concentrated sulfuric acid
3.4.1.2 Apparatus and machines
 Six 25ml volumetric flasks
 1000ml volumetric flasks
 Measuring cylinders
 Funnel
 UV 1609PC UV/VIS Spectrophotometer
 Glass beakers
 Standard calibration curve for Nitrates
3.4.2 Reagents preparation
All the chemical reagents used will be analytical grade and the water used will be distilled water

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Sodium salicylate solution 0.5g Na- salicylate will be dissolved in 100ml distilled water, this
solution will be stable for one day only
Sodium hydroxide tartrate solution, the solution will be made by dissolving 400g of NaOH in 1
liter of distilled water together with 50g of sodium tartrate
3.4.3 Stock and working solution preparation
A stock solution will be prepared by dissolving 6.067g of sodium nitrate in 1 liter of distilled
water to make a stock solution of 1000mgl-l
From the stock solution, a working solution with concentration of 5mg Na- No3 per liter will be
made by taking 5ml of stock solution and diluting to 1 liter
Standard series of concentrations was made by taking the given concentrations of the working
solution and diluting with distilled water as shown in the table below;
Working solution (Mg) Volume of distilled water(Ml) Concentration (Mg/l)
0.0 20.0 0.0
1.0 19.0 0.25
2.0 18.0 0.5
4.0 16.0 1.0
10.0 10.0 2.5
20.0 0.0 5.0

3.4.4 Procedure for Nitrate analysis

 20ml of filtered water sample will be placed in an evaporation bottle
 To this as well to the standard series 1ml of sodium salicylate solution freshly prepared
will be added
 The bottles will be then put in the oven and the sample will be dried at a temperature of
95⁰c
 The results will be dissolved quantitatively by adding 1ml of concentrated sulfuric acid
and then the bottles will swirled careful while still warm
 40ml of distilled water will be then added to it and mixed
 Finally 7ml of sodium hydroxide tartrate solution will be added, mixed and then the
absorbance determined at a wavelength of 420nm

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 CHAPTER FOUR
4.0 Results and discussion of results
Determination of maximum value of Nitrates
Wavelength (√). Absorbance (Au)
O.0 0.0
100 0.050
200 0.100
340 0.300
360 0.327
380 0.430
400 0.530
420 0.536
440 0.390
460 0.207
480 0.112
500 0.074
Highest peak for a given concentration of standard is determined at different wavelength
4.1 Graph of highest absorbance for Nitrates

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Lambda max for Nitrates
Concentration and absorbance value for Nitrates
Concentration (mg/l). Absorbance (Au
0.0 0.177
0.25. 0.182
0.50. 0.139
1.00. 0.280
2.50. 0.536
5.00. 0.821
The values are used in plotting a standard curve
4.1.1 Graph for Nitrates standards

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4.1 .1.1 Calibration curve for Nitrates
4.1.2 Table for concentration of Nitrates in the stream samples
Rivers Up stream mid-stream lower stream average
Kipsongo 6.03_+0.26. 5.57_+0.16. 5.37_+0.04. 5.90-+0.08
Lovington 3.72_+0.17. 3.99_+0.12. 4.02_+0.40. 3.90_+0.09
Kimana 6.34_+0.16. 6.02_+0.18. 5.53_+0.11. 6.16_+0.40
R.Athi 4.30_+0.15. 4.70_+0.21 4.00_+0.20. 4.33_+0.07
4.2 Discussions
 Kipsongo river
Nitrates concentration range is between
This pollution is attributed to the fact that the river has its sources in Kitale which is highly
industrialized, most of the industrial effluents find their way into the river
 Lovington stream
Nitrates concentration range between
These concentrations are below the recommended level of 50mg/l by WHO, major activities
upstream were livestock grazing and watering, domestic water use and irrigation farming
 Kimana swamp
Nitrates concentration was almost constant for the first sampling sites at
The concentration declined to
At the flow termination point. These concentrations are below the recommended level of
50mg/l by WHO
 River arthi
Water samples were collected at the spring where it emerged from underground Shanti
center and at two other points, the Nitrate content at the spring was at
The Nitrate concentration in the irrigation water discharge was
These concentration are below the recommended level of 50mg/l by WHO, laundry,
livestock watering and other domestic uses are the factors that contribute to the level of
Nitrates in the river.
4.3 Discussion of beer Lambert’s law and its application in the research
The method used to ensure the concentration of pollutants in in the water is carried
spectrophotometry which is a procedure that determines how much a chemical compound
absorbs and transmits light, having the light absorbance profile of a solution we can compare it
with a known sample and identify which compound are present and also their quantities
Spectrophotometry is commonly used in analysis of water sample which was explored in this
experiment, it measures the intensity of light that passes through a solution and comparing the
intensity with reference blank. Spectrometers are only able to emit and detect light within a

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specific wavelength range and are classified accordingly, A common type is the uv-visible
Spectrophotometer that emits and absorbs photons with wavelength in the ultraviolet
range(185nm to 400nm) and visible range (400nm to 700nm) while another type , the IR
Spectrophotometer uses wavelength in the infrared range (700nm to 1500nm) the most
accessible of these types is the visible light Spectrophotometer which was used in this
experiment. The solutions color can be used in an indicator of the solutions light absorptivity and
transmissivity certain chemical compound absorbs light at specific wavelengths unique to the
system.
If the solute is present in a higher concentration more light will be absorbed as described by the
Beer Lambert’s equation or Beer’s law, this expression relates the absorbance to the solute
concentration in a given solution.
A=elc(l)
Where
A. Is the compounds absorbance (unit-less)
e. Is the molar absorptivity [l(mol/cm)]
L. Is the path length (cm)
C. Is the compound concentration (mol/l)
In order to obtain quantitative absorption data a set of standards with known concentration must
be analyzed in order to create a list of values to compare with the unknown sample
4.4 Basic instrumentation of a Spectrophotometer
Light source
Collimator
Monochromator (prism of grating)
Slit (wavelength selector)
Cuvette (sample solution)
Detector (photo cell)
Digital display

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4.5 Application of Beer Lambert’s law
The analysis is based on the reaction of Nitrate with sodium salicylate in a sulfuric acid
medium, which form yellow colored salts of nitro salicylic acid. The intensity of the
yellow color is determined spectrophotometric ally using the Beer Lambert’s law
Laboratory analysis for Nitrates ions.

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 CHAPTER FIVE
5.0 Conclusion and Recommendations
5.1 Conclusions
 From the research, it is concluded that the stream water in Kipsongo, Kitale do have
Nitrates though lower than the WHO recommend level of 50mg/l
 Nitrates in stream water of Kipsongo, Kitale pause health hazards to residents who use it
for drinking and cooking
5.2 Recommendations
From this study it is recommended that:-
 Education on safe handling of fertilizers and application procedures to reduce water
pollution by Nitrates needs be intensified
 Health ministry need to raise level of awareness on the health hazards paused by water
polluted with Nitrates.

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 LIST OF REFERENCES
Chapra SC, Pelletier GJ 2008) QUALZK a modeling framework for simulating River and stream
Becky H.D (2013) principles of field ionization and field desorption mass spectrometry,
international series in analytical chemistry. Elsevier
Babora mikendova, Jan Thomas, Thomas Danek (2010) “fluorescence spectrophotometry and
stand spectrophometric method” Geoscience engineering 6(4) pp 22-31
Conway, Gordon Ramsay and Jules N. Pretty unwelcome harvest agriculture and pollution
Routledge 2013
Berner EK and Berner R.A (2012) Global environment, water, air and geochemical cycles.
Princeton university press

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 APPENDIX

Mass spectrometry, international series in analytical chemistry. Elsevier

Water analysis
Parameter – nitrate
Health hazards paused by nitrates in water

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