Running Head: CHEMISTRY 1

Chemistry

Student Name

Instructor Name

Date
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Chemistry

Colourimetry

Colorimetery is a chemical method used to determine the concentration of a colored compounds

in a solution or its a technique that measures the concentration of a known constituent of a

solution by comparison with colors of a standard solution of that constituent (Colorimetry 1998).

It can also be determined as the physical intensity which is used to determine the concentration

of a colored compound in a solution.

The device used to measure the concentration of a colored compound is called Colorimeter. It is

a light-sensitive device and work by measuring the transmittance and absorbing of light passing

through a liquid sample. It measures the intensity or concentration of the color that develops

upon introducing a specific reagent into a solution.

Two types of Colorimeters used to measure the concentration One is Densitometers and the

other is Color photometers (Sapan, Lundblad and Price 1999).

Densitometer: It measures the intensity of primary colors.

Color photometers: It measures the color reflection and transmittance.

Colorimeter has three components, these are a light source, a cuvette containing the sample

solution and a photocell attached for detecting the light that pass through the solution. It is also

equipped with either colored filters or specific LEDs to generate color. The output is displayed

by an analog or digital meter in terms of absorbance or transmittance in Colorimeter.

Principle of Colorimetry
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According to Beer Lamberts Law, Absorption of light transmitted through the medium is

directly proportional to the concentration of the medium.

The principle of colorimeter is based on this law. Working of the device is that a beam of light

with a specific wavelength through a series of lenses is passed through a solution, which

navigates the colored light to the measuring device. It analyses the color of the solution

compared to an existing standard. Calculation of absorbance or percentage transmittance is

calculated by the microprocessor. More light will be absorbed if the concentration of the solution

is greater, which can be identified by measuring the difference between the amount of light at its

origin and that after passing the solution.

Several sample solutions of a known concentration are first prepared and tested, in order to

determine the concentration of an unknown sample. The concentrations are then plotted on a

graph against absorbance that generates a calibration curve. The results of the unknown sample

are compared to that of the known sample on the curve to measure the concentration.

Method

1. A 0.002 mol. /L paracetamol solution has been prepared to use as a stock solution for your

practical.

2. To each of 7 test tubes add 1.0 ml HCl (6 mol. /L) and 2 ml sodium nitrite solution (10%;

freshly prepared and pre-made).

3. Label the tubes carefully and then add stock paracetamol solution and water to the tubes as

indicated.
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Tube 1 2 3 4 5 6 7

Paracetamol 0 0.2 0.4 0.6 0.8 1.0 0

(ml)
Water (ml) 1.0 0.8 0.6 0.4 0.2 0 0

Additionally, add 1.0 ml of the test solution (unknown concentration) to tube 7.

4. Mix all the tubes thoroughly and allow them to stand for 2 minutes.

5. Add 2 ml ammonium sulphamate (15%) drop wise to each tube. Do this carefully to avoid

frothing.

6. Add 2.5 ml sodium hydroxide (25%) to each tube and mix for 15 seconds.

7. Allow the tubes to stand for 2 minutes to allow any bubbles to disperse. Gently tapping the

side of the tube may help in this respect.

8. Measure the absorbance of tubes 2-7 at 430 nm using tube 1 as the blank. If bubbles form in

the cuvettes these should be dispersed with a glass rod as they will lead to inaccurate colorimeter

readings.

Result

Tube no Concentration Absorbance

1 0 0
2 0.0004 0.133
3 0.0008 0.302
4 0.0012 0.42
5 0.0016 0.535
6 0.002 0.691
7 Unknown 0.325
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0.8
0.7
0.6
f(x) = 0.08x + 0.01
0.5
R = 0.59
0.4
Absorbance
0.3
Linear ()
0.2
0.1
0
0 1 2 3 4 5 6 7 8

Concentration

Discussion

Many analytical methods are used to determine plasma concentration of Paracetamol in

biological fluids, Colorimetry with chemical derivatization is one of them. It is essential to

determine/measure the concentration of Paracetamol in biological fluids because of the need to

acquire antidote therapy, driven by the needs of clinical toxicology requiring the rapid, reliable

and highly specific estimation of Paracetamol in plasma samples. However, paracetamol is a

widely available over the counter drug and cheap to buy and is a common drug used in overdose.

It has an excellent analgesic and antipyretic properties. Due to these sufficient reasons and its use

in overdoses, plasma concentration in vivo is studied and measured (Shihana, et al. 2010).
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References

Acetanilides: Advances in Research and Application: 2011 Edition. Atlanta: ScholarlyEditions,

2012.

Colorimetry. July 20, 1998. https://www.britannica.com/science/colorimetry (accessed February

23, 2017).

Patel, HV, and DJ Morton. "Specificity of a colorimetric paracetamol assay technique for use in

cases of overdose." Journal of Clinical Pharmacy and Therapeutics 13, no. 3 (1988):

233-238.

Rainsford, Kim D. Aspirin and Related Drugs. Boca Raton: CRC Press, 2016.

Sapan, CV, RL Lundblad, and NC Price. "Colorimetric protein assay techniques." Biotechnology

and Applied Biochemistry, 1999: 99-108.

Shihana, Fathima, Dhammika Menike Dissanayake, Paul Ivor Dargan, and Andrew Hamilton

Dawson. "A modified low-cost colorimetric method for paracetamol (acetaminophen)

measurement in plasma." Clinical Toxicology 48, no. 1 (2010): 42-46.