Module-3

Instrumental methods of analysis:

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 Colorimetry

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Colorimeter refers to a device used in colorimetry that
aids in the absorption of a particular wavelength of light
by a specific sample solution.
Louis J. Duboscq invented it in the year 1870. It is employed
to measure how much light transmits and absorbs as it
passes through a liquid. These are used to identify the color
and establish the concentration of a solution. By comparing a
solute’s color intensity in a solution to that of a reference
solution with a known solute concentration, one can estimate
the concentration of colored solute in that solution.

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Principle of Colorimeter
When an incident light beam with intensity I0 passes through a solution, a part of the
incident light is reflected (Ir) and absorbed (Ia) while the remaining incident light is
transmitted (It).
i.e., Io = Ir + Ia + It
The measurement of (I0) in the colorimeter eliminates (Ir), and it is sufficient to
calculate the (Ia). Using cells with the same characteristics maintains a steady amount
of light reflection (Ir). Then (I0) & (It) are measured.
The operation of the colorimeter is based on Beer-Lambert’s law which states that the
amount of light absorbed by a color solution is directly proportional to the solution’s
concentration and the length of a light path through it.
A ∝ cl
A = ∈cl
Where,
A = Absorbance/ Optical density of the solution
∈ = Coefficient of absorption
c = Concentration of solution
l = Length of the path
A series of lenses in a colorimeter guide a beam of light with a particular wavelength
through a solution as it makes its way to the measuring apparatus. This compares the
colour to a current standard to examine it. The absorbance or % transmittance is then
calculated using a microprocessor. By measuring the difference between the amount
of light at its source and that after passing the solution, it is possible to determine20the
Several sample solutions with known concentrations are first prepared and
evaluated to ascertain the concentration of an unknown sample. Plotting the
concentrations versus absorbance on a graph yields the calibration curve. To
determine the concentration, the results of the unknown sample are contrasted with
those of the known sample on the curve.

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Instrumentation of Colorimeter
Light Source: The source of light should produce energy with enough intensity to cover
the entire visible spectrum (380-780 nm). Commonly, Tungsten lamps are used as a light
source for measurement in the visible spectrum and near-infrared ranges. Halogen
deuterium is suitable for measurement in the UV range (200-900 nm).
Slit: It reduces unwanted or stray light by allowing a light beam to pass through.
Condensing lens: Parallel beam of light emerges from condensing lens after the light
passes through slit incidents on it.
Monochromator: It filters the monochromatic light from polychromatic light, which
absorbs unwanted light wavelengths and permits only monochromatic light. These are of
three types: prism, grating, and glass.
Prism: It facilitates the refraction of light when it passes from one medium to another.
Glass: It selectively transmits light in certain ranges of wavelengths.
Gratings: These are made of graphite, which separates light in different
wavelengths.
Cuvette (Sample cell): The monochromatic light from the filter passes through the
colored sample solution placed in the cuvette. Their sizes range from square, and
rectangle to round and have a fixed diameter of 1cm. These are of three types based on
the substances these are made of: Glass, Quartz, and Plastic cuvette.
Glass cuvettes are cheap and absorb light of 340 nm wavelength.
Quartz cuvettes facilitate entry of both lights of UV and visible ranges.
Plastic cuvettes are cheaper, easily scratched, and have shorter lifespans.
Photocell (Photodetector): These photosensitive devices measure light intensity by
converting light energy into electrical energy.
Galvanometer: The electrical signal generated in a photocell is detected and measured 22
by a galvanometer. It displays optical density (OD) and percentage transmission.
Types of Colorimeters
Several types of colorimeters are as follows:
Densitometers: Determine the density of a material.
Spectrophotometers: Measure the spectral reflectance and
transmittance of a surface.
Tristimulus colorimeter: Employed to measure the tristimulus
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values of a color.
Colorimeter Operating Procedure
[Link] the device on by rotating the Power Switch knob in a clockwise
direction (toward the right). 15 minutes of warming up time for the
colorimeter is required to stabilize the light source and the detector.
[Link] the warm-up period, turn the Wavelength Control knob to the
appropriate wavelength.
[Link] the MODE control key until the light next to “Transmittance” turns on
to switch the display mode to transmittance.
[Link] the Zero Control knob to set the display’s T-factor to 0.0%. Make this
adjustment while ensuring the sample chamber is empty and the cover is
securely closed.
[Link] the blank solution in a cuvette until it reaches the top of the triangle
on the side of the cuvette. To get rid of any fluids or fingerprints on the
cuvette’s exterior, wipe it with a Kimwipe. Both will obstruct the light’s ability
to travel and result in inaccurate readings.
[Link] the tube gently but completely into the cuvette chamber, with the
vertical guide line facing in the direction of your right. The guideline on the
cuvette should now be lined up with the guideline on the sample chamber by
rotating the cuvette 90 degrees in a clockwise orientation. This method
protects the cuvette against scratches in the light-transmitting portions.
Erroneous measurements can result from scratches on the cuvette. Close
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the compartment’s cover.
Colorimeter Operating Procedure
[Link] the display to 100.0% using the Transmittance/Absorbance
control knob.
[Link] the MODE control key and switch the Status Indicator light to
read Absorbance. The display should indicate 0.0 if the
Transmittance calibration was done correctly. No further adjusting is
necessary. Use the Transmittance / Absorbance control knob to set
the display to 0.0 if it does not already show that value. Switch the
display back to Transmittance using the MODE key.
[Link] the previous process to remove the cuvette from the
compartment by rotating it 90 degrees counter-clockwise before
doing so. You should put the solution whose absorbance you want to
test in another cuvette. Similar to before, place it inside the chamber.
10.a. Directly from the digital display, read the %T value.
b. Select Absorbance using the MODE key, then take the A value
directly from the digital display. Select Transmittance once more.
11. Reverse the process you used to insert the cuvette to remove it
from the sample compartment. Close the compartment’s cover.
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Applications of Colorimeter
[Link] is employed in the printing industries to evaluate the caliber of printing
ink and paper.
[Link] are used in the food and food processing industries.
[Link] is frequently used in laboratories and hospitals to determine the
biochemical composition of samples like blood, urine, cerebral spinal fluid,
plasma, serum, etc.
[Link] is used in the textile and paint industries.
[Link] dealers also examine the visual characteristics of priceless
stones using a colorimeter.
[Link] instrument is also employed in cosmetology to measure the UV
protection level of skin-care products.
[Link] are used to evaluate the water’s purity and screen for the presence
of chemicals like cyanide, iron, fluorine, chlorine, molybdenum, etc.
[Link] are employed to evaluate the color contrast and brightness of
screens on mobile devices, computers, and televisions to give people the
greatest viewing experience.
9.A colorimeter is also employed in the pharmaceutical sector to spot inferior
goods and medications.
[Link] samples are tested using a colorimeter to determine the amount of
hemoglobin present. 26
Advantages of Colorimeter
1.A quick and affordable means of evaluating quality is the
colorimeter.
[Link] Colorimeter makes it simple to perform a quantitative
examination of colored chemicals.
[Link] are available in under a second.
[Link] AA batteries can be measured using a portable
colorimeter between 100 and 300 times.
Disadvantages of Colorimeter
[Link] procedure of determining the concentration of colorless
substances becomes laborious.
[Link] does not function in the ultraviolet or infrared
spectrum since it only measures wavelength absorbance in the
visible range of light (400nm to 700nm).
3.A spectrum range must be set rather than a specific
wavelength to measure the absorbance.
[Link] might be challenging on surfaces that reflect
light. 27
 The percolation of water through soil and rock dissolves minerals like Iron and
Manganese in that and takes along with its flow. Water containing iron and
manganese is actually colourless when it is not in contact with the oxygen. But when
the same water is exposed to air, it gets oxidized causing change in water colour.
The oxidation of dissolved iron particles in water changes the iron to white, then to
yellow and finally to red-brown solid particles that settle out of the water. Iron that
does not form particles large enough to settle out and that remains as colloidal iron
leaves the water with a red tint. Although manganese is present in water in a
dissolved form it leaves black tint which is due to the colloidal manganese in
shallow wells. This water containing Iron is one of the minerals required by the
human body. It is used in the production of the oxygen-carrying proteins,
haemoglobin and myoglobin. A deficit of iron in the body can leave a person feeling
tired and listless, and can lead to a disorder called anaemia. Presence of iron in water
is not harmful for health, but it renders the water turbid, imparts colour to water and
hence unacceptable from the aesthetic point. It causes satins in laundries and
plumbing fixtures. Its presence may help in the proliferation of iron reducing
bacteria in the aquifer and the pipe lines. Hence, the desirable limit is 0.3mg/l in
water and the tolerable limit is 1.0mg/l.
To detect the presence of these ions in solution, thiocyanate ions (SCN -) are
added. Thiocyanate ions respond with iron (III) ions in solution to form a deep red
coloured complex ion: Fe3+(aq) + SCN-(aq) → [FeSCN]2+(aq)
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or, more fully, [Fe(H2O)6] (aq) + SCN (aq) → [Fe(H2O)5SCN] (aq) + H2O (l)
3+ - 2+
colours of a series of standard solutions, with known
Fe3+ concentrations, the concentration of iron in the food sample
may be determined. This method is called colorimetry. Colorimetric
analysis is based on the change in the intensity of the colour of a
solution with dissimilarities in concentration. Colorimetric methods
represent the simplest form of absorption analysis. The human eye
is used to associate the colour of the sample solution with a set of
standards until a match is found. An increase in sensitivity and
accuracy results when a spectrophotometer is used to measure the
colour intensity. Mainly, it measures the fraction of an incident
beam of light, which is transmitted by a sample at a particular
wavelength.
Phenanthroline Spectrophotometric Method :
This method relies on the fact that iron, when converted to its
ferrous state by reaction with acidified hydroxylamine, forms a
colour-stable complex with phenanthroline. The red coloured
complex formed can then be measured with a spectrophotometer
at a wavelength of 510 nm. There are two ways to measure the
difference in intensity of the light beam. One is the percent
transmittance, %T, which is defined as:
Relevant Indian Standard for Total Iron Test on Water :
[Link] 3025 (Part 53) -2003: Method of Sampling and Test (Physical29
[Link]
enivronmental_science_engineering_laboratory_met
hodology/
determination_of_iron_and_manganese_in_water.p
hp#bb

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Methods of determining the trace organic and
inorganic contaminants using emission and
absorption technique

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MelroyCastalino/organic-and-inorganic-
trace-contaminates-detection-using-aas

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Emission and absorption technique.
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