ABSTRACT

In this experiment, it was aimed to determine the quantity of Chromium, Cr (VI) concentration
present in a polluted lake water sample by using spectrophotometer and to perform the proper
method of preparing a series of standard solutions between 1 to 100 ppm that need to be used in
the calibration curve of spectrophotometer thus to analyze whether the lake water sample tested is
suitable for drinking water and/or agricultural purposes. Basically, the experiment was carried out
by dilution method of preparing five sample of 20mL standard solution having different
concentration of Chromium (VI) which were 15ppm, 30ppm, 45ppm, 60ppm and 75ppm. The
series of standard solution were tested in spectrophotometer and the absorbance value obtained
were recorded. A calibration curve for Cr (VI) was plotted. Next, the lake water sample collected
at Taman Tasik Shah Alam was tested in the spectrophotometer. The absorbance value of the lake
water sample was 0.103. Then, the concentration of Cr (VI) present in the water sample was found
to be 72.33ppm. It was higher than the maximum acceptable value standardized by Ministry of
Health Malaysia which is 0.05 ppm. Therefore, the lake water is contaminated and thus is not
suitable for drinking water or agricultural purposes.

1
 INTRODUCTION

Many heavy metals, including Chromium, Cr are toxic even at low aqueous solution
concentrations. Trivalent Cr (III) compounds are not usually considered as health hazards but
hexavalent Cr (VI) compounds can be toxic if ingested or inhaled and have been established as
carcinogens. Chromium ions, Cr (III) or Cr (VI) are naturally found in the rivers, lakes and streams.
Basically, naturally alkaline rivers and lakes have much lower risk of such contamination of
chromium ions compared to the naturally acidic water streams. Since the Cr (VI) is a strong
oxidizing agent and poses a high toxicity to humans and animals, the determination of Cr (VI) in
environmental and biological samples is of great interest.

In general, food appears to be the major source of chromium intake. Drinking-water intake
can, however, contribute substantially when total chromium levels are above 25 g/litre. In terms
of environmental effects, chromium enters air, water and soil in the form of Cr (III) and Cr (VI).
Various activities that human had been done that increase the concentration of chromium such as,
chemical, textile and leather manufacturing as well as electro painting.

Cr (III) is an essential human dietary element and occurs naturally in many vegetables,
fruits, meats, grains and yeast. It is a nutritionally essential element in humans and is often added
to vitamins as a dietary supplement. Normally, Cr (III) has relatively low toxicity and would be a
concern in drinking water only at very high levels of contamination. Cr (VI) occurs naturally in
the environment from the erosion of natural chromium deposits but it can also be produced by
industrial processes.

There are demonstrated instances of chromium being released to the environment by

leakage, poor storage, or inadequate industrial waste disposal practices. Cr (VI) is more toxic and
poses potential health risks. People who use water containing total chromium in excess of the
maximum contaminant level (MCL) over many years could experience allergic dermatitis.
Therefore, Cr (VI) is classified as likely to be carcinogenic to humans when ingested.

2
 OBJECTIVE

The objective of this experiment is to determine the quantity of Chromium, Cr (VI)

concentration present in a polluted lake water sample by using spectrophotometer and to perform
the proper method of preparing a series of standard solutions between 1 to 100 ppm that need to
be used in the calibration curve of spectrophotometer thus to analyze whether the lake water
sample tested is suitable for drinking water and/or agricultural purposes.

THEORY

Chromium is a naturally occurring element. However, elemental chromium Cr (0) does not
appear in nature but it is found complexed with oxygen, iron or lead. Chromium can exist in nine
different oxidation states, from -II to +VI, but the common valence states are +II, +III and +VI.
Because of their stability in the environment, the trivalent Cr (III) and hexavalent Cr (VI) forms
are the most common.

Basically, Cr (III) is considered to be the most thermodynamically stable oxidation state

under ambient redox conditions. It is a positively charged ion that has a strong tendency to form
hexacoordinate octahedral complexes with a variety of ligands (oxygen, nitrogen or sulphur
atoms). These stable complexes can prevent the precipitation of Cr (III) above pH 5-6, where it
would otherwise precipitate.

In other hand, Cr (VI) is not thermodynamically stable. It is a strong oxidizing agent,

existing only as tetrahedral oxo species such as chromium oxide (CrO3), chromyl chloride
(CrO2Cl2) and chromate ion (CrO42-). This is produced during the reduction of chromite ore to
obtain chromium metal. Cr (VI) in the environment comes from anthropogenic pollution where Cr
(VI) rarely occurs naturally due to its affinity for organic matter and other reducing substances. In
solution, Cr (VI) exists as an anion and quite mobile in the environment; the dissolved species of
Cr (VI) are hydrochromate (HCrO-), dichromate (CrO2-) and chromate (CrO42-).

3
 Generally, common source of chromium contamination in natural waters is automobiles
due to its component in various automobile breaks as well as parts of engine. Chromium that
normally deposited on highroads because of brake dusts and exhaust of automobile engine can
unfavorable diffuse into any water streams nearby via flowing rainwater. Therefore, lakes and
water streams which located near a typical congested with automobiles area are the most
susceptible areas to be contaminated by such chromium. Its cancer-risk character and toxicity even
at mere concentrations has attracted a lot of federal environmental agencies to do researches in
determining its presence in many environmental samples. Thus, in this experiment absorption
spectroscopy will be used to detect the concentrations of Chromium (VI) in a water sample.

During this experiment, absorption spectroscopy will be used to detect the concentrations
of Chromium (VI) in a water sample. In point of fact, colored aqueous solutions have chemical
species contents which able to absorb significant wavelengths of light. Similarly, heavy metals can
be identified via absorptions of wavelengths of light. The amount of light absorbed is linearly
proportional to the concentration of metal ions.

Absorption spectroscopy operates on the measuring principle of light before and after it passes
through aqueous metal solutions. Basically, the amount of light absorbed by the chemical species
in the sample is equivalent to the difference in amount of light before it enters the sample and after
it exits the sample. Since the light is to be absorbed by chemical species, it must be set to a specific
wavelength. Every chemical species absorbs distinct wavelength of light. In absorption
spectroscopy, the wavelengths of light absorbed by a metal in solution are detected.

In this experiment, standard solutions will be prepared by diluting 300 parts per million of
Chromium (VI) standards solution. Five standard solutions need to be prepared from the
concentrated Chromium (VI) standard solution. To calculate the dilute solutions volume, the
solution dilution formula can be used, as what us written below:

(M1)(V1) = (M2)(V2) ..(1)

4
Quantitative analysis using spectrophotometer is based on Beer-Lambert Law:

A= MLC(2)
Where,

A = absorbance value (dimensionless, thus no units is represented)

M = molar absorbance (L/mol.cm)
L = path length of the cuvette in which the sample is contained (cm)
C = concentration of the compound in solution (mol/L)

Beer-Lambert Law states that absorbance value depends on the total quantity of the absorbing
compound in the light path through the cuvette. Thus, if plot a graph of absorbance versus
concentration of the compound solution, a straight line passing through the origin (0,0) is
determined. The molar absorbance, M, is a constant for a particular substance, therefore if the
concentration of the solution is halved, so is the absorbance value. A compound with a high molar
absorbance is very effective at absorbing light (of the appropriate wavelength), and hence low
concentration of a compound with a high molar absorbance can be easily detected.

5
 PROCEDURES

1. All the materials and apparatus were setup.

2. The standard solutions of chromium were prepared with the different concentrations which
were 15 ppm, 30 ppm, 45 ppm, 60 ppm and 75 ppm.
3. By using the pipette, 1 mL, 2 mL, 3 mL, 4 mL and 5 mL of chromium (VI) solution were
taken and placed in five different 20 mL volumetric flasks respectively and was labeled.
All the preparations were conducted in the fume hood.
4. Each volumetric flask was filled with distilled water until it reaches the mark on the
volumetric flask indicates the volume of 20 mL.
5. The volumetric flask was closed with the stopper and then inverted for almost 20 times or
more to ensure that the solution mixed well.
6. The spectrometer was switched on.
7. 10 mL of distilled water was filled into the cuvette and placed in the spectrometer slot.
Before that, no bubbles were ensured in the cuvette because it can affect the reading of the
absorbance.
8. The zeroing step was done on the spectrophotometer to ensure that all data obtained were
accurate.
9. A small portion of each 5 series of diluted solution which known as standard solution, was
transferred into the cuvette and placed in the spectrometer slot. Before that, the cuvette was
ensured to be clean from dirt and also fingerprint to avoid from error reading of absorbance
of the solution.
10. The reading of absorbance value for each solution was recorded and tabulated into a data.
11. A calibration curve of the standard solution was plotted based on the absorbance values
and the concentration of Cr (VI).
12. The lake water sample was filled into the cuvette and placed in the spectrometer slot to get
the reading of absorbance.
13. All the data was recorded and analyzed.

6
 APPARATUS

20 mL volumetric flasks, pipette, 250 beaker, dropper, conical flask, spectrophotometer, 10 mL

square cuvette bottle.

MATERIALS

300ppm of standard solution of Chromium (VI) solution, distilled water, lake water sample.

RESULTS

Volume of chromium solution used (mL) 1 2 3 4 5

Concentration of chromium 15 30 45 60 75
Absorbance value 0.054 0.064 0.074 0.085 0.109
Table 1: Determination of absorbance value at different concentration of Chromium (VI)
solution using spectrophotometer

Lake water sample Absorbance value

1 0.103
2 0.103
3 0.103
Average 0.103
Table 2: Determination of absorbance value of water sample using spectrophotometer

7
 The calibration curve of Cr (VI)
0.12

0.11
y = 0.0009x + 0.0379 0.109
R = 0.9562
0.1

0.09

0.085
0.08

0.074
0.07
Absorbance Value

0.064
0.06

0.054
0.05

0.04

0.03

0.02

0.01

0
0 10 20 30 40 50 60 70 80
Concentration of Cr (VI)

Graph 1: The calibration curve of absorbance value versus concentration of Chromium (VI).

8
 SAMPLE OF CALCULATIONS

To prepare diluted 20 mL of standard solution of 15 ppm Cr (VI) concentration, the solution

dilution formula is used to calculate the volume of Chromium solution (VI).

M1V1= M2V2

(300 ppm)(V1) = (15 ppm)(20mL)

V1 = 1.0 mL

The same method of calculation was performed for the other solution:

30 ppm M1V1= M2V2

(300 ppm)(V1) = (30 ppm)(20mL)

V1 = 2.0 mL

45 ppm M1V1= M2V2

(300 ppm)(V1) = (45 ppm)(20mL)

V1 = 3.0 mL

60 ppm M1V1= M2V2

(300 ppm)(V1) = (60 ppm)(20mL)

V1 = 4.0 mL

75 ppm M1V1= M2V2

(300 ppm)(V1) = (75 ppm)(20mL)

V1 = 5.0 mL

9
The equation of the linear line was given by as:

y = mX + C

Based on the plotted graph, the equation of linear line obtained is:

y = 0.0009x + 0.0379

By substituting the value of y with the average of absorbance value of lake water sample, which
is 0.103 in the equation given as:

0.103 = 0.0009x + 0.0379

Hence, we get that the value of x is:

0.1030.0379
x=
0.0009

x = 72.33

Generally, x is equivalent to the concentration of Chromium (VI) present in the lake water sample
according to the graph which is 72.33 ppm.

10
 DISCUSSIONS
This experiment was carried out to complete the objectives, which is to determine the
concentration of Chromium VI which present in simulated lake water sample by using a
spectrophotometer. Another objective is to use a method of dilution to prepare solutions of
concentration of Cr (VI) of 15, 30, 45, 60, 75, and 100 ppm. Another objective is to analyze the
water samples if they are suited for drinking purpose and agriculture purposes. The lake water
sample is believed to be contaminated as the lakes is situated near roadway and industrial zones.
As mentioned before, lake and water stream situated in those locations have high risk to contain
heavy metals such as chromium and generally is classified as contaminated water source.

In this experiment, the parent solution of Cr (VI) solution is set at 300 parts per million
concentration. 5 series of dilute solutions are prepared at fixed volume of 20 mL. To prepare a
solution of concentration of 15 ppm, 1.0 mL of Cr (VI) solution is used and mixed with distilled
water. In other hand, 2.0 mL of Cr (VI) solution is used for 30 ppm, 3.0 mL of Cr (VI) solution is
used for 45 ppm, 4.0 mL of Cr (VI) solution is used for 60 ppm and 5.0 mL of Cr (VI) solution is
used for 75 ppm. The absorbance for each of the standard solution is tested by using
spectrophotometer. The graph represent the calibration curve of Cr (VI) is then plotted.

According to the graph, a simple linear regression analysis is performed to determine the
linear best fit for the graph of absorbance value versus concentration data. The R2 value indicates
how well the regression analysis fits the absorbance-concentration data. The closer the R2 value
to1.00, the better the linear regression analysis has fit the data.

The absorbance value of lake water sample is taken by using the spectrophotometer. Based
on the linear equation of the graph, the calculation of determining the concentration of Chromium
(VI) present in the lake water sample is performed. The value of concentration of Cr (VI) in the
lake water sample calculated is 72.33 ppm. Based on Ministry of Health Malaysia, the maximum
acceptable value for chromium to present in drinking water as well as raw water is 0.05 ppm.
Therefore, since the lake water sample has higher concentration of Cr (VI), it is unsafe and
unhygienic for drinking water thus cannot be used for public consumption or even agriculture
purposes.

11
 CONCLUSIONS

The concentration of Chromium (VI) present in the collected lake water sample tested is
72.33 ppm. It is higher than the maximum acceptable value standardized by Ministry of Health
Malaysia which is 0.05 ppm. As the conclusion, the lake water is obviously contaminated and thus
is not suitable for drinking water or agricultural purposes.

RECOMMENDATIONS

There are some recommendations that can be suggested to avoid doing mistakes during the
experiment. First and foremost, when measuring small volume of chromium (VI), always ensure
that the most accurate apparatus is used which is a pipette. The volume inserted inside the pipette
can be controlled more accurately rather than using measuring cylinders. Then, when using the
pipette, our eyes must be perpendicular to the meniscus to avoid parallax error. During the dilution
process, the volumetric flask must be shake and invert several times to ensure that it mixed well.
In fact, the selection of wavelength of light in the spectrophotometer is quite essential. Therefore,
to obtain an ideal value, it is preferred to use a wavelength where the component absorbs
substantially.

Next, the cuvette bottles must be cleansed and dried properly with a distilled water in order
to avoid inaccurate contents of either standard solutions or the chromium solution. In addition, the
cuvette bottles must be held properly. Avoid leaving any fingerprints or smudge imprinted on the
outer surface of the cuvette that will unfavorably become such interference for the solutions to
absorb wavelengths of light in the spectrophotometer. Therefore, the person that handling the
bottles must wear a pair of gloves throughout the experiment. In addition, this experiment must be
repeated thrice to obtain more accurate value of the absorbance.

12
 REFERENCES

(2015). Retrieved 1 November 2015, from

http://www.who.int/water_sanitation_health/dwq/chemicals/chromium.pdf

(2015). Retrieved 2 November 2015, from http://www.gunungganang.com.my/pdf/Malaysian-

Policies-Standards-
Guidelines/Standards/National%20Drinking%20Water%20Quality%20Standard.pdf

Docslide.us,. (2015). Determination of Chromium (VI) Concentration via Absorption

Spectroscopy Experiment - Documents. Retrieved 1 November 2015, from
http://docslide.us/documents/determination-of-chromium-vi-concentration-via-absorption-
spectroscopy-experiment.html

Docslide.us,. (2015). Exp 3 Determination of Cr (VI) Concentration via Absorption Spectroscopy

- Documents. Retrieved 1 November 2015, from http://docslide.us/documents/exp-3-
determination-of-cr-vi-concentration-via-absorption-spectroscopy.html

Healthycanadians.gc.ca,. (2015). Chromium in Drinking Water. Retrieved 1 November 2015, from

http://www.healthycanadians.gc.ca/health-system-systeme-sante/consultations/chromium-
chrome/document-eng.php

Law.resource.org,. (2015). Indian Standard: METHODS OF SAMPLING AND TEST (PHYSICAL

AND CHEMICAL) FOR WATER AND WASTEWATER. Retrieved 2 November 2015, from
https://law.resource.org/pub/in/bis/S02/is.3025.52.2003.html

Scribd,. (2015). Determination of Chromium (VI) Concentration via Absorption Spectroscopy

Experiment. Retrieved 1 November 2015, from
http://www.scribd.com/doc/39166236/Determination-of-Chromium-VI-Concentration-via-
Absorption-Spectroscopy-Experiment#scribd

Scribd,. (2015). determination of concentration of chromium. Retrieved 1 November 2015, from

http://www.scribd.com/doc/110293635/determination-of-concentration-of-chromium#scribd

13
Water.epa.gov,. (2015). Basic Information about Chromium in Drinking Water | Basic Information
about Regulated Drinking Water Contaminants | US EPA. Retrieved 1 November 2015, from
http://water.epa.gov/drink/contaminants/basicinformation/chromium.cfm

Wikipedia,. (2015). Atomic absorption spectroscopy. Retrieved 1 November 2015, from

https://en.wikipedia.org/wiki/Atomic_absorption_spectroscopy

14