Topic 2- Questions

1. Spectroscopy is the study of the interaction between ___________ and _________

(fill in the gaps)

2. In UV-vis Spectroscopy why do we use Absorbance instead of Transmittance for

quantitative analysis?

3. Beers low is A = bc, where

A = ______
= ______
b = ______
c = ______
(fill in the gaps)

4. How would you define a calibration curve?

5. IR region can be divided into:

_______: 0.8-2.5 m
_______: 2.5-15 m
_______: 15-1000 m
(fill in the gaps)

6. Mid-IR Spectroscopy is mostly applied to ________ analysis, whilst Near-IR is mostly

for ________ analysis.
(fill in the gaps)

7. Briefly describe the Infrared Milk Analyser

8. Chromatography is a separation techniques based on the ___________ of a sample

(solute) between a ________phase and a _______ phase.
(fill in the gaps)

9. List 3 different chromatographic modes

10. List 3 kinds of liquid chromatography

11. List 3 advantages of HPLC over classic liquid chromatography (LC)

Answers: all answers to above questions can be found in Topic 2 slides

Topic 2- Calculations
Problem 1. A colored radical cation has a molar absorptivity of 2.1 103 cm -1M-1 at 514 nm.

(a) What will be the absorbance of a 1 10-4M solution in a 10 mm cuvette at 514 nm?
(b) What will be the percent transmittance of the solution in (a)?

Solution: (a) 0.21; (b) 61.6%

Problem 2. In the analysis of the antioxidant capacity of one red wine and one white wine
sample (three replicates for each sample) using DMPD radical cation and Trolox as standard
antioxidant, the following data were obtained:

%T at 514 Abs at
sample I514% TEAC value* Trolox calibration curve
nm 514 nm
31.0

39.3
White wine
33.5
y = 0.7466 x + 4.3419
(where y = I514%;
44.7
x = Trolox g/ml)
51.4
Red wine
56.0

Calculate:
- the Absorbance of the samples at 514 nm from the %T (percent Transmittance)
- the I514(%) from the equation:

I514(%) = [1- (As / A0)] 100, considering A0 = 0.702 and A0 = Abs at 514 nm

- the TEAC values (Trolox Equivalent Antioxidant Capacity) in g/ml from the Trolox
calibration curve taking into account the following dilution factors:

- White wine: diluted 1:20

- Red wine: diluted 1:100

Decimal places required:

Absorbance: 3
I514(%): 1
TEAC values in g/ml: 2

Solution: TEAC for white vines: 620.36 g/ml; 1014.15 g/ml; 748.94 g/ml.
TEAC for red vines: 6128.86 g/ml; 7294.15 g/ml; 8004.03 g/ml.