RESULT

SAMPLE ABSORBANCE CONCENTRATION

NUMBER

STD 1 0.003 0.000

STD 2 0.195 5.000

STD 3 0.557 15.00

STD 4 0.919 25.00

STD 5 1.221 35.00

STD 6 1.640 45.00

50

45 45
0

40

35 35
0

30

25 25
0

20

15 15
0

10

5 5
0

0 0 0 0 0 0 0
1 2 3 4 5 6

ABSORBANCE VS CONCENTRATION
DISCUSSION

Ultra-visible spectrophotometer used in this experiment was to determine the maximum

wavelength of Carmoisine solution. Carmosine is another synthetic food dye in the red to
maroon colour range. It is in the azo dye group.

In this experiment, the stock solution of 100ppm Carmosine is diluted into the 5 serial
dilution which is 5ppm, 15ppm, 25ppm, 35ppm, and 45ppm. 2 unknown solution was from
the mixed of 2 serial dilution into one and did the same way for the second unknown. The
blank solution the distilled water was used during analyse when using UV-VIS
spectrophotometer.

The cuvettes used in this experiment was the most important to be taken care of. The
cuvette has 2 different surfaces, where the rough ones can be touched by bare fingers and
the other ones, which are the smooth ones shouldn’t be touched by fingers. This is because
the smooth sides of the cuvette are where the light will go through the sample from the
source. If the smooth sides of cuvette were stick with fingerprints, the light might be
diffused to another way. That was why wiping the smooth surfaces of the cuvette is very
important. There were two types to determine in λmax max scanning which is wavelength
scanning and photometric scanning. To determine the Carmosine sample, 45ppm was firstly
used to scan because it contain high of concentration. For obtain the photometric scan,
each dilution was scanned to produce calibration graph.

Based on the result, when the concentration was 0.000 the absorbance is the lowest which
is 0.003. But when the concentration is 45.000 the absorbance is the highest which is 1.640.
It can be concluded that the higher the concentration, the value absorbance also will
become higher.

The data that were obtained consists of concentration values of five the standard solution
with their absorbance and standard calibration graph. It also consists of standard deviation.
The concentration of the unknown sample also obtained. And the concentration of
unknown also were determine. But although the concentration of unknown solution has
been obtained by instrument, manual calculation still been done for comparison. After
obtaining the results, the linear calibration graph was re-plotted using Microsoft Office
Word software . The value of R2 that was obtained from the graph is 0.9987. Based on the
data, it can be concluded that the result is closely to the value of theory that is 1. The
machine that were used in this experiment is SHIMADZU 80109.

There were few errors that might occur during conducting the experiment that might
affect the result. Firstly, the distilled water was over from the marked level when added into
the round bottom flask that contained Carmosine solution. It will affect the concentration of
Carmosine solution.
Wavelength Scan

(i) Instrument Parameters

Starting Wavelength = 400 nm

Ending Wavelength = 800 nm

Path length = 10 mm

(ii) Result for Wavelength Scan

Sample used = Carmosine

Concentration of the sample = 45 ppm

max obtained = 514.5 nm

C. Photometric Scan

(i) Instrument Parameters

Wavelength,  = 514.5 nm

Path length = 10 mm

(ii) Results for Photometric Scan

Record the absorbance readings and the unknown concentrations in the table below.

No. Sample Concentration (ppm) Absorbance

1 Std 1 5 0.195

2 Std 2 15 0.557

3 Std 3 25 0.919

4 Std 4 35 1.221

5 Std 5 45 1.640

6 Unknown #1 15+5 0.423

7 Unknown #2 45+35 1.494

a. By referring to the standard calibration graph obtained, the r2 value is 0.9987