Determination of Nitrate in Drinking Water by UV

CHM 4101 Lab Report 1.
GOKHUL, Raja.
October 30, 2015.
1015485.
Group 1: Alicia Sukhdeo, Leanna Simon and Shivraj Balkisson.
Title:
Determination of Nitrate in Drinking Water by UV

Spectrophotometry.
Aim:
To Determine Nitrate in Drinking Water by UV

Spectrophotometry.
Theory:
The UV spectrophotometer can perform electronic analysis

on the solutions (calibration standards, unknowns and control) and
in so doing yield quantitative information pertaining absorbance,
that is, the amount of a specific wavelength of light/
electromagnetic radiation that an analyte absorbs.
The UV spectrophotometer has a few basic components.
These include the source, a device for wavelength selection, the
sample compartment and detector. Considering this study, the
source would most likely be a Deuterium lamp since this lamp is
capable of producing a continuous UV emission with wavelengths
ranging from 185 nm to about 375 nm which is satisfactory for
most UV analysis. Next is wavelength selection, this may be
achieved via absorption filters and a monochromator. From here a
specific wavelength of light, like 340 nm will be produced. This
specific wavelength is then feed to the sample compartment which
holds the sample. The analyte will absorb an amount of this light
that is proportional to the concentration of the analyte. The
unabsorbed light goes on to the detector where the signal is
amplified and can then be quantified.
From the beer lambert law we know that absorption is
proportional to the concentration of the sample and the shape/size
of the sample container. This is why a standard sized and shaped
cuvette is used as the sample container. Another thing to consider
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October 30, 2015.
1015485.
is that components of the matrix may also absorb light. In our case
this is corrected by using a double-beam spectrophotometer. In this
spectrophotometer, the light coming from the device that selects
the wavelength which came from the source is split via beam
splitter or light chopper so that two daughter beams are produced.
One of the daughter beams goes on to the matrix while the other
goes on the sample. As such any absorbance by the matrix can be
subtracted from that of the sample to get the absorbance of the
analyte only.
In this case, the analyte is nitrate. Nitrate has max at the
wavelengths of electromagnetic radiation 220 nm and 275 nm.
However, because of the probable presence of the interference,
dissolved organic matter, in the solution, a correct is made and the
absorbance is taken at 340 nm and 395 nm instead of 220 nm and
275 nm.
By making solutions in which the concentration of Nitrate
is known to a high level of accuracy, that is, 0.5, 1.0, 3.0, 5.0 and
10.0 ppm Nitrate and then taking the absorbance of these five
systems, one can construct a standard curve. From the standard
curve, a straight line would be obtained. This line will have an
equation given by = + (equation 1). In this equation, m
and c will be known constants (calculated via MS word 2013), y
represents the absorbance and x represents the concentration of
Nitrate in ppm in the water. Therefore by measuring the
absorbance of the unknown solutions and the control solution, the
concentration of Nitrate in ppm in these solutions can be
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October 30, 2015.
1015485.
calculated simply via the use of a derivative of equation 1 which is

given by equation 2; =
According to the Environmental Protection Agency (2015),

the maximum contaminant level for nitrate that is allowed in
drinking water is 10 ppm. Moreover, Saunders (2014) verifies that
lead nitrate is an ingredient of paint.
Diagram:
The spectrophotometer that utilized in this laboratory

Exercise is shown in figure 1.
Figure 1: Diagram of the UV-Vis Spectrophotometer

Method:
As per Laboratory Handout barring the following

amendments;
1. Absorbance for each solution was read at 340 nm and 395 nm
instead of 220 nm and 275 nm.
2. 50 mL beakers were used as an alternate to overcome the
shortage of 50 mL volumetric flasks.
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October 30, 2015.
1015485.
Data:
Please see Appendix 1.
Graph:
Treatment of Data:
Results:
The results obtained from Laboratory Exercise 1 for group

1 is tabulated in table 1.
Table 1: Results obtained from Laboratory Exercise 1 for group 1.
Concentration of Nitrate in solutions /
Solutions
Unknowns
ppm
1
44.0
66.0
83.0
0.0
Control
Discussion:
A perfect standard curve was not obtained from the

absorbance of the five calibration standards. The experimenter
believes that this could be due to a few reasons. Such as;
The intensity of the light varying for each run for

the calibration standards. Even if one calibrating
standard (5 ppm) has more nitrate than another
calibrating standard (3 ppm) and the intensity of the
light source is lower in the for the 5ppm run than
the 3ppm, then the expected higher absorbance for
the 5 ppm solution because it has more analyte to
absorb more light, would not be observed. This is
not unreasonable as the UV-Vis spectrophotometer
is quite old and would be prone to such faultiness.
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October 30, 2015.
1015485.
The introduction of interferences into the sample by

the experimenters since they did not wear gloves for
the entire experiment. As such, even if one
calibrating standard (3 ppm) has less nitrate than
another calibrating standard (5 ppm) and nitrate,
especially from the hands of the experiments enter
the sample then, the expected lower absorbance for
the 3 ppm solution because it should has less
analyte to absorb less light, would be reversed and
the expected outcome would not be not be
observed.
The use of an old spectrophotometer that may be

very well be defective. Suppose the detector of the
instrument has lost it functionality over the years,
then the absorbance readings would not be accurate
and therefore anomalies appear in the study like this
one. With samples having more analyte (5 ppm)
absorbing less than samples with less analyte (3
ppm).
All three of the unknown solutions had nitrate
concentrations above the maximum contaminant level

of the EPA (10 ppm). This was expected since the
unknown solution in question was taken from a site that
is very close to a facility that does painting. Paints
contain lead nitrate as confirmed via Saunders (2014).
It would not be a stretch of the imitation to conclude
that paint or its vapour that contain the aforementioned
nitrate gets into the water via diffusion.
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October 30, 2015.
1015485.
On average the drinking water (the unknown

solution in question) that was obtained from next to a
facility that does painting had a nitrate content of 64
ppm. This exceeds the EPA limit by over 500 % ,
therefore it is unfit for human consumption.
As it relates to why the same unknown had
variances of roughly 20 ppm nitrate for each run, the
experimenters believes the reasons given below would
be able to explain;
The use of an old spectrophotometer that may be

very well be defective.
o Suppose the intensity of the light varying for
each run for. Even if all solutions have the
same concentration but the intensity of the
light source is lower in one run (like
unknown 1) than another run (like unknown
2) then unknown 1 would absorb less light
and therefore have a smaller absorbance
than unknown 2.
o Suppose the detector of the instrument has
lost it functionality over the years, then the
absorbance readings would not be accurate
and therefore anomalies appear in the study
like this one. With samples having the same
concentration of analyte have different
absorbance.
The introduction of interferences into the samples

of each run by the experimenters since they did not
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October 30, 2015.
1015485.

wear gloves for the entire experiment. As such,
even if all the samples should have the same
concentration and absorb the same amount of light
and have the same absorbance, each sample has
vastly different absorbance because each sample
was exposed to different amounts of interferences.

The control solution would not be useful in
these cases because it does not contain the
interferences from the experimenters, thus, these
interferences cannot absorb any light, contribute to
the controls absorbance which would be subtracted
from that of the sample. All it has is the matrix from
the sample.
The reason for adding HCl to all of the
systems studied was for it to act as a solvent in this
study.
Conclusion:
It can be concluded that the drinking water

obtained from next to the facility that does painting
has a Nitrate content of 64 ppm on average.
Therefore it is unfit for human consumption via
EPA standards.
References:
1. Kenkel, J. (2014). UV-Vis and IR Molecular Spectrometry. In Analytical chemistry for
technicians (4th ed.). Boca Raton: Lewis.
2. Clark, J. (2006). Double beam UV-visible absorption spectrometer. Retrieved January 24,
2016, from http://www.chemguide.co.uk/analysis/uvvisible/spectrometer.html#top
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October 30, 2015.
1015485.
3. Unite States Environmental Protection Agency. (2015, December 8). Table of Regulated
Drinking Water Contaminants. Retrieved January 24, 2016, from
http://www.epa.gov/your-drinking-water/table-regulated-drinking-watercontaminants#Organic
4. Saunders, N. (n.d.). Making Paints. Retrieved January 24, 2016, from
http://www.creative-chemistry.org.uk/activities/paints.htm
5. Thermo Spectronic [PDF]. (n.d.). Retrieved January 24, 2016, from
http://www.molecularinfo.com/MTM/UV.pdf
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CHM 4101 Lab Report 1.

October 30, 2015.

Group 1: Alicia Sukhdeo, Leanna Simon and Shivraj Balkisson.