JOMO KENYATTA UNIVERSITY OF AGRICULTURE AND TECHNOLOGY.

Name: Wambugu Florence Waithira.

Reg. No: SCP 212-0378/2016.

Course: BSc. Industrial Chemistry.

Unit Code: SCH 2455.

Unit Name: Applications of Analytical Chemistry.

Lecturer: Dr. Anam Onditi.

Report: Determination of Nitrates in JKUAT Water.

Date: 26th November 2019.

DETERMINATION OF NITRATES IN JKUAT LABORATORY TAP WATER USING
UV/VISIBLE SPECTROPHOTOMETER.

Objectives.

1. To quantitatively determine the amount of nitrates in the JKUAT laboratory tap water.
2. To determine the effectiveness of sewage treatment works.

Literature Review.

Nitrate is commonly monitored for environmental protection purposes in agricultural, food

controls, the control of multiple uses of water and also in checks its potability. Owing to the
formation of carcinogenic N-nitrosamines and being also essential to indicate organic pollution
in water, the determination of the exact concentration of nitrate is desirable. In recent years, an
increasing interest in the determination of nitrate levels in food products has been observed,
essentially due to the potential reduction of nitrate to nitrite, which is known to have adverse
effects on human and animal health. Nitrate is intermediate states in the nitrogen cycle, and the
high concentration of nitrite in water generally indicates poor quality. Presently, nitrate can be
quantified in plasma, serum and urine samples by various methods based on different analytical
principles. These methods have produced diverging values of nitrate in the circulation of healthy
humans. This variation can only partially be explained by differences in dietary intake of these
compounds. The major reason for the discrepancies seems to be methodological problems. A
stable metabolite of NO is nitrate and it is generally accepted as an indicator of NO. This indirect
measurement is generally used because NO has a very short half-life in biological systems. Most
studies of NO concentrations from vitreous samples are conducted by detection of nitrate using
the Griess reaction. Nitric oxide (NO) partly auto-oxidizes to nitrite (NO2-), whereas by far the
major endogenous and exogenous sources fraction of NO is oxidized to nitrate (NO3-) within the
erythrocytes by oxyhemoglobin (HbFe(II)O2). Nitrate levels from vitreous samples are quantified
by the chemical reduction of nitrate to nitrite, and then the reaction of nitrite with the Griess
reagents produces a purple azo dye which can be measured at 540 nm.

Many spectrophotometric methods are available for the determination of nitrate. Extensive use
has been made of diazotization and coupling reaction after reduction of nitrate to nitrite, copper-
coated cadmium metal and titanium (III) chloride have been used. However, reduction should
not proceed beyond nitrite. Reduction of nitrate to nitrite and the catalytic effect of nitrite on the
oxidation of Naphthol Green B by bromate, or reduction to nitric oxide and chemiluminescence
detection by reaction with ozone have also been used.

Among the various methods in preconcentration and determination, solid-phase extraction has
received more acceptances due to its simplicity, rapidity and attainability of large
preconcentration factor. A variety of solid materials such as modified ion exchange
resins, functionalized resins with chelating reagents, modified nanometer-sized alumina, thermal
modified Kaolinite, activated carbon, zeolites, cellulose, and immobilized microorganisms on
sepiolite have been used for preconcentration of trace metals. Microcrystalline naphthalene and
benzophenone have also been used as solid phase for adsorptive extraction of metal ion
complexes.

When determining nitrates, it is important to choose a method which suits both the interferences
which are present and the level of analyte in the solution. The UV-Visible method of determining
nitrate is a useful method for screening of a solution to make a rough determination of the level
of nitrate. It determines the absorption of the nitrate ion at 220nm. Good results are obtained if
the water contains little or no organic matter (which also absorbs at 220nm). A correction for
organic matter can be performed by taking absorbance readings at 275nm (where nitrate does not
absorb) and estimating the organic content of the sample. After screening the sample and
determining the amount of nitrate other methods may be chosen and used to give more accurate
estimates of nitrate content of the water sample. For example, the nitrate ion selective electrode
is suitable from 0.14 – 1400mg/L. In this practical, the nitrate ion level of JKUAT tap water is
determined by screening the sample with the UV-Visible method.

Materials and Reagents.

1. Analytical weigh balance

2. Volumetric Flasks
3. Pipettes
4. UV-Visible Spectrophotometer.
5. Cuvettes.
6. Autoclave.
 7. Sodium nitrate
8. Distilled Water
9. Laboratory Tap water
10. 1+15 HCl.
11. 1+500 HCl.

Procedure.

1.371g of sodium nitrate was dissolved in 1000ml of distilled water to yield 1000ppm of nitrate
ions. 100ppm of the nitrate ions was thereafter prepared in 100ml volumetric flask and thereafter
2ppm, 4ppm, 6ppm, 8ppm and 10ppm standards of nitrate ions were prepared from the 100ppm
in 100 ml volumetric flasks. To each of the prepared standards, 5ml of 1+500 Hydrochloric acid
was added and the standards were set aside for 15 minutes after which they were ready for
analysis. 10ml of tap water was collected, filtered and topped up to 50ml using distilled water.
Afterwards, 5ml of 1+15 hydrochloric acid was added to the tap water and was allowed to stand
for 15 minutes. The absorbance readings of the standards and the sample were recorded at
220nm.

Calculations.

1. From the results, plot the calibration curve and obtain the actual concentration of nitrates
in the tap water.
 Absorbance Curve of Nitrates
0.5

0.45 f(x) = 0.04 x + 0.02

R² = 0.99
0.4

0.35

0.3
Absorbance

0.25

0.2

0.15

0.1

0.05

0
0 2 4 6 8 10 12

Concentration in ppm

The concentration of the nitrates in the water x = (y – 0.021)/ 0.0438

X = (0.102 – 0.021)/0.0438
X= 1.8493
Actual concentration = (X × Dilution Factor × Volume Used) / Mass
Taking the water density as 1g/ml;
(1.8493 × 1 × 10ml)/10g
Actual Concentration of nitrates = 1.8493ppm

Discussion.

The normal limit of nitrate content in clean water according to the National Environmental
Management Authority of Kenya is 50ppm and in drinking water is 10ppm. Nitrate in drinking
water can be harmful to infants and young children, lead to Methemoglobinemia and increased
risk of spontaneous abortion or certain birth defect on pregnant women if it exceeds 10ppm. The
effect of high nitrate concentration can also cause ovarian and bladder cancer, cardiovascular
disease, stomach cancer, miscarriage, high blood pressure, the formation of nitrosamine and bad
effect on the nervous system. Spectrophotometric method was often used to nitrate determination
because this method was simple and sensitive. The actual concentration of nitrates in the tap
water from the experiment was found to be 1.8493ppm. This concentration was within the
required concentration of nitrates (< 10ppm) for both clean and drinking water with regards to
the water regulations by NEMA. Consequently, this implied that the waste water treatment for
the tap water JKUAT was using was properly treated and the water was safe for use. The
experiment had few human errors that were as a result of parallax errors while measuring
volumes and also loss of volumes while transferring the materials in different flasks. The errors
presence are evident in the regression value of 0.986. The errors however did not cause a huge
variation on the results. Hence, the experiment was a success.

Conclusion.

The actual concentration of the nitrates in the tap water was found out to be 1.8493ppm. This
was within the nitrate concentrations of clean drinking water stated in the water regulations act
of Kenya. Therefore, the water was well treated at the sewerage treatment plant. All the
objectives were met and thus, the experiment was a success.

References.

1. http://www.cffet.net/envan/W6.pdf
Retrieved on 26th November 2019 at 1233 hours.
2. https://pubs.acs.org/doi/abs/10.1021/ac60202a036
Retrieved on 26th November 2019 at 1245 hours.
3. http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-40422011000400010
Retrieved on 26th November 2019 at 1300 hours.
4. https://aip.scitation.org/doi/pdf/10.1063/1.5016005
Retrieved on 26th November 2019 at 1313 hours.
5. https://www.tsijournals.com/articles/direct-determination-of-nitrate-in-natural-water-by-
ultraviolet-first-derivative-spectrophotometry.pdf
Retrieved on 26th November 2019 at 1337 hours.