Members: Catangay, Marie Antoinette P, Date Performed: Sept.

15, 2017
Esporlas, Juan Miguel Date Submitted: Sept. 22, 2017
Section: 3ChEB Group No.: 4
Experiment No. 5
AN INTRODUCTION TO ULTRAVIOLET-VISIBLE SPECTROSCOPY

I. Treatment of Results

1) The relationship between the wavelength and absorbance

Table 1. Wavelengths of Peak Maxima of the 0.25 M Solutions

Solution (0.25M) Peak Maxima Wavelength (nm)
Cu2+ solution 6.998 776
Ni2+ solution 1.214 394
Fe3+ solution 8.533 392
Co2+ solution 1.171 512

Figure 5. Spectra of the 0.25M Solutions

2) Absorbance vs Wavelength (assigned ions ~ Nickel and Cobalt)

Cobalt Solution
0.7
0.6
Absorbance

0.5
0.4
0.3 y = 4.9364x
R² = 0.9903
0.2
0.1
0
0 0.02 0.04 0.06 0.08 0.1 0.12 0.14
Actual Concentration

Figure 7. Line Equation of the Cu2+ Solutions (w.r.t. expected concentrations)

 Nickel Solution
0.8
0.7 y = 4.565x + 0.0839
0.6 R² = 0.9935

Absorbance
0.5
0.4
0.3
0.2
0.1
0
0 0.02 0.04 0.06 0.08 0.1 0.12 0.14
Actual Concentration

Figure 8. Line Equation of the Ni2+ Solutions (w.r.t. expected concentrations)

Table2. List of Actual Concentrations of the Diluted Cu2+ Solutions

Cobalt Solution
Expected Concentration Absorbance @ 512 nm Actual Concentrations
0.125 0.635 0.125
0.0625 0.279 0.063
0.031 0.137 0.031
0.0156 0.084 0.016
0.0075 0.063 0.008

unknown solution 0.319 0.064

Table 3. List of Actual Concentrations of the Diluted Ni2+ Solutions

Nickel Solution
Expected Concentration Absorbance @ 394 nm Actual Concentrations
0.125 0.668 0.125
0.0625 0.347 0.063
0.031 0.213 0.031
0.0156 0.165 0.016
0.0075 0.136 0.008

unknown solution 0.402 0.0609

 I. Spectrophotometer Data
Cu2+ Ni2+ Fe3+ Co2+
SOLN SOLN SOLN SOLN

WL (nm) ABS ABS ABS ABS

350 0.079 0.133 9.197 0.052

400 0.027 1.165 7.544 0.091
450 0.024 0.114 1.655 0.534
500 0.026 0.015 0.728 1.113
550 0.068 0.04 0.151 0.526
600 0.242 0.159 0.042 0.1
650 0.755 0.436 0.039 0.075
700 1.709 0.491 0.055 0.039
750 2.655 0.446 0.078 0.024
800 2.434 0.23 0.094 0.033

II. Serial Dilution Computation

Serial Dilution (M1V1=M2V2)
M1 (M) V1(ml) M2 (M) V2 (mL)
0.25 7.5 0.125 15
0.125 7.5 0.0625 15
0.0625 7.44 0.031 15
0.031 7.55 0.0156 15
0.0156 7.21 0.0075 15

II. Answer to Guide Questions

1. What is the significance of 𝛌𝑚𝑎𝑥 of a spectrum?

The 𝛌𝑚𝑎𝑥 indicates the wavelength used to determine the absorbance of the unknown
solutions. The peak maxima expresses this situation and shows the maximum fraction of light that the
solution can absorb.

2. How did you decide on the wavelength at which you will read the absorbance of your diluted
solutions? What will be the effect of choosing about 20 nm lower or higher than your chosen
wavelength?
 The wavelength chosen was the one with the highest peak, especially for solutions with more
than one peak. For the reason that choosing the highest peak will be much easier to see because it is
visible and observable when the solution was scanned. Choosing the data with lower than 20nm or
higher than the wavelength will make the data unreliable and not accurate. Recording the data must be
accurate enough because small changes or details may be seen in your graph and it will be harder to
interpret.
3. Compare the absorbance readings of the cuvet with and without the finger prints. What is the
significance of the finger print experiment?
There is a big difference between a cuvette with fingerprints and a clear cuvette. Obviously, the
clear cuvette is the efficient and accurate than the one with fingerprints because fingerprints can be a
hindrance to the light that passes through and the source of error on the readings.

4. What are the other uses of the spectrophotometer?

The spectrophotometer is not limited only for photometry and spectrum readings. It has lots of
advantages and uses. It can be used in forensics, medical purposes, experiments, and material science.
It is one of the most useful methods of quantitative analysis in various fields such as chemistry, physics,
biochemistry, material and chemical engineering, and clinical applications. Qualities of
semiconductors, fingerprints, and DNA can be identified with the help of spectrophotometer.

5. Why do you have to use the same cuvet after doing the baseline correction?
In the experiment proper, the diluted solutions were tested all together labeled as A, B, C, D,
and E with the same cuvette with distilled water in the autozero container in the spectrophotometer, so
that even the data gathered will still be accurate even if the performer changes the wavelength where
the device will read.
6. What are the other uses of the spectrophotometer?
It can also be used in different fields such as forensics and materials science. Qualities of
semiconductors can be identified with the help of spectrophotometer. DNA (deoxyribonucleic acid) and
fingerprints of a person can also be identified with such device [4]
A spectrophotometer is used in a variety of areas of science such as microbiology, biochemistry, forensics, physi
and medical health.
It can be used to measure certain ingredients in a drug to make sure it is effective and safe for consumers. You ca
measure bacterial growth, or diagnose a patient based on how much uric acid is present in their urine.
In Astronomical Spectrometry, we can determine the wavelength of a body, which can then give us their chemica
composition (as a factor of their spectra and mass), temperature, distance and speed (using a function of
their wavelength and the speed of light), starting from simple spectroscopic analysis of a body, and also used to
identify the chemical makeup of objects in space.
Another type is Mass Spectronomy. Once the particles are separated, they’re measured by an electron multiplier
and we can identify the makeup of the sample through the weight of each ion’s mass.

7. Why do you have to use the same cuvet after doing the baseline correction?

Remember to use the same cuvet which was used for baseline correction so that even the data gathered
will still
be accurate even if the performer changes the wavelength where the device will read

Each cuvet, although assumed similar, are not identical. Changing the cuvet would affect the path length (l) in
the Beer's Law, thus affecting the absorbance readings.