VISUAL LAB WORKSHEET – AUTUMN 2020-21

Name: Sugondo Limin

Student ID: 20113828
Group No: 3

Please submit this worksheet to the assessor on duty by 12pm or 4pm.

Results and Discussion

Results

Wavelength determination: ___________269.10___________ nm

Table 1: Calibration data.

Standard solution Concentration (ppm) Absorbance
0
Blank 0
0.5069
A 10
1.0128
B 20
1.5508
C 30
2.1057
D 40
5.7205
E 100

Table 2: Concentrations of caffeine in samples.

Set A: without dilution
Beverage sample Average Concentration (ppm) Possible type of beverage*

103.07
U1 Pressed coffee
30.75
U2 Decaffeinated coffee
100.85
U3 Black tea
100.11
U4 Green tea
104.28
U5 Instant coffee

Set B: 10 times dilution

Beverage sample Average Concentration (ppm) Possible type of beverage*

5.00
V1 Green Tea
3.08
V2 Decaffeinated Coffee
V3 5.34 Black Tea

1
 8.39
V4 Pressed Coffee
9.55
V5 Instant coffee

Set C: 50 times dilution

Beverage sample Average Concentration (ppm) Possible type of beverage*

1.64
W1 Black Tea
1.23
W2 Decaffeinated Coffee
1.61
W3 Green Tea
2.20
W4 Pressed Coffee
2.40
W5 Instant Coffee

*It was known that the samples were sourced from the following beverages: (1) instant coffee,
(2) black tea, (3) green tea, (4) decaffeinated coffee, and (5) pressed coffee. Estimate the type of
beverage according to the concentration determined. Support your decision with literature data.
From USDA: https://fdc.nal.usda.gov/

Calculations on standards preparation:

M1V1=M2V2
Set B (10 times dilution):
Dilution factor= Volume of diluted solution/ Volume of concentration solution
10= Volume of diluted solution/ 100mL
Volume of diluted solution= 1000mL

Set C (50 times dilution):

Dilution factor= Volume of diluted solution/ Volume of concentration solution
50= Volume of diluted solution/ 100mL
Volume of diluted solution= 5000mL

Discussion

1. What are the parameter settings in the “Method” section?

X=wavelength
X start at 800nm, end at 190nm
Y=absorbance
SBW = 20nm
Spectra No = 1
Data Interval = 1 nm
Scan Rate = 240 nm/min

2. Comment on the calibration graph’s linearity:

2
 R2 value of the graph is 0.99698. This value is very close to 1, indicating that the calibration
graph is almost linear. Therefore, the graph is suitable and reliable to be used to predict the
concentration of unknown samples.

3. How certain are you about the accuracy of the calibration data? If you prepare the standards in
different concentrations (as opposed to those used in the experiment), would you expect the
calibration graph to be different?

The calibration graph shows that the results is in compliance with the Beer's law. Beer’s law
relates the concept of concentration and absorbance. Therefore, the calibration data is highly
accurate. In accordance to this law, absorbance and concentration are directly proportional. If
the original concentration is increased, then the absorbance will increase and if the diluted
solution has a reduction in the original concentration, hence the absorbance will decrease in
direct proportion. Therefore, the calibration graph will also show direct proportional trend.

4. How confident are you about the exact type of beverage in the samples? Use statistical
analysis to support your argument. In addition, compare your results with literature and
comment.

Black tea (20mg/100g) and green tea (11mg/100g) have smaller differences in concentration
in the literature, as observed from the data in Set A, B and C. The caffeine concentration in
green tea (11mg/100g) is very much higher than decaffeinated coffee (1mg/100g). Similar
trend is observed in Set A, B and C

5. What is the effect of dilution factor variation on the readings obtained? Would you expect any
discrepancies?
The dilution factor will affect the concentration of the solution relating to M1V1=M2V2.
As a solution is diluted which reduce the concentration, the absorbance of the solution is
decreased. This is due to absorbance is directly proportional to concentration of the solution. If
a 1.0 L of a 1.0 molar solution is diluted to 2.0 molar solution, the new molar concentration is
0.50. The new concentration will be reduced by one half as the volume was doubled.

6. What are the possible sources of error? Comments in terms of statistical deviations. How can
these errors be overcome? What are your recommendations to achieve the most efficient
analysis?

Error and how to overcome:

-environmental effects on photometer and sample error can be overcome by keeping the
photometer and sample clean.
-line voltage fluctuations error can be overcome by reducing the change of reactive power in
the supply system.
- sample solutions are not homogeneous, mixing it well prior to experiment.
- bubble occurrence

Achieve the most efficient analysis

- Ensure that sample-isolation protocols are optimized, and that the samples are purified prior
to measurement
- Ensuring that both top and bottom measurement surfaces are clean prior to loading blanks
or samples is important to avoid problems with subsequent readings
- Avoid introducing bubbles when pipetting the samples.

7. What other analytical method would you suggest for the same purpose of measuring the
caffeine concentrations? How would you decide which is the most suitable analytical
technique to use?
Atomic Absorption Spectroscopy (AAS). Refer to the linearity of calibration curve produced by both
UV-Vis and AAS. The graph that yields a greater R 2value will indicate the more suitable and reliable
analytical technique.