Experiment No.

1
UV-Spectrophotometer
Objective:
To measure the concentration of a given analyte in water using UV-VIS spectrophotometer.

Group # 4

Moosa Naseer 2016-CH-17

Rehan Jami 2016-CH-37
Syed Sher Ali Bukhari 2016-CH-27
Mubarram Ali 2016-CH-45

Description:
• Calibration by using distilled water.
• Maximum wavelength.
• Analysis of the sample.

Procedure:
• First of all, we have to switch ON the equipment by pressing start button.
• After that it is required to prepare a sample set consisting of aqueous ferric chloride
solutions of four different concentrations of:
1. 100 ppm
2. 200 ppm
3. 500 ppm
4. 1000 ppm
• Now, we have to take 4 quartz rectangular cuvettes and fill them with distilled water up to
atleast 75 % of total length.
• First, we have to set-up the equipment by giving it a maximum wavelength to work on i.e.
635 nm for ferric chloride solution.
• After then, press F4 for entering the interface of configuration option for blank test.
• Put the quartz tube filled with distilled water into the sample box inside the left chamber
of UV-spectrophotometer.
• Press F1/F2/F3/F4 so that all four quartz cuvettes filled are analyzed by equipment. Press
enter and take out all cuvettes.
• Now, drain out all the distilled water inside the cuvettes and fill all the cuvettes with one
by one with each of solutions of defined compositions.
• Press start and then all the results will be plotted on the interface which will be Absorbance
vs. Conc.
 • It is to be noted that there will be one curve each for one concentration at set-wavelength
of equipment.

Observations and Calculations:

673

673

Graphical Representation:

Ferric oxide solution conc. (Steeper) Vs Phenolic iron conc (Horizontal)

Conclusion:
The UV-Spectrophotometer present in environmental engineering laboratory, however, being an
ancient machine have plotted the absorbance vs. conc. in a good visual demonstration but results
were much inaccurate due to problems in equipment’s operating case in the path between where
the lights enters and where it is transmitted. The equipment was suffering optical hindrance with
keystrokes response slight slower than what should be observed.

Future prospect of UV-Spectrophotometer Experiment:

The primary understanding of using the UV-spectrophotometer was related to the concept of
behavioral analysis of optical response of the instrument with changing composition of solution.
The calibration curve have helped us to understand the deviations of predictive results from
experimental results. Thus, it will help us to determine any impurity in water and thus, allowing
us to take better precautionary measures in coping with the environmental issues, thus promoting
green engineering co1. Detection of Impurities.

Significance of UV-Spectrophotometer:
UV absorption spectroscopy is one of the best methods for determination of impurities in organic
molecules. Additional peaks can be observed due to impurities in the sample and it can be
compared with that of standard raw material. By also measuring the absorbance at specific
wavelength, the impurities can be detected.

Structure elucidation of organic compounds.

UV spectroscopy is useful in the structure elucidation of organic molecules, the presence or
absence of unsaturation, the presence of hetero atoms.
From the location of peaks and combination of peaks, it can be concluded that whether the
compound is saturated or unsaturated, hetero atoms are present or not etc.

Quantitative analysis
UV absorption spectroscopy can be used for the quantitative determination of compounds that
absorb UV radiation. This determination is based on Beer’s law which is as follows.
A = log I0 / It = log 1/ T = – log T = abc = εbc
Where ε is extinction co-efficient, c is concentration, and b is the length of the cell that is used in
UV spectrophotometer.

Qualitative analysis
UV absorption spectroscopy can characterize those types of compounds which absorbs UV
radiation. Identification is done by comparing the absorption spectrum with the spectra of known
compounds.