Solution: KKEK2233 Physical and Analytical Chemistry II

Lecturer: Dr. Awanis

Tutorial 3 Date: 8th May 2018

1. Ethanol and methanol are separated in capillary GC column with retention times of 370 and 385
s, respectively, and base widths (wb) of 16.0 and 17.0 s. An unretained air peak occurs at 10.0s.
Calculate the separation factor and the resolution. (6 marks)

𝟑𝟖𝟓 𝟐
𝑵 = 𝟏𝟔 ( ) = 𝟖. 𝟐𝟏 𝒙𝟏𝟎𝟑 𝒑𝒍𝒂𝒕𝒆𝒔
𝟏𝟕
𝟑𝟖𝟓 − 𝟏𝟎
𝜶= = 𝟏. 𝟎𝟒
𝟑𝟕𝟎 − 𝟏𝟎
𝟑𝟕𝟎 − 𝟏𝟎
𝒌𝟏 = = 𝟑𝟔. 𝟎
𝟏𝟎
𝟑𝟖𝟓 − 𝟏𝟎
𝒌𝟐 = = 𝟑𝟕. 𝟓
𝟏𝟎
𝟑𝟔𝟎 + 𝟑𝟕. 𝟓
𝒌𝟐𝒂𝒗𝒆 = = 𝟑𝟔. 𝟖
𝟐

𝟏 𝟏. 𝟎𝟒𝟐 − 𝟏 𝟑𝟕. 𝟓
𝑹𝒔 = √𝟖. 𝟐𝟏 𝒙 𝟏𝟎𝟑 ( )( ) = 𝟎. 𝟗𝟏
𝟒 𝟏. 𝟎𝟒 𝟑𝟔. 𝟖 + 𝟏

2. Substances A and B were found to have retention times of 6.4 min and 14.4 min, respectively,
on a 22.6 cm column. An unretained sample of air passed through the column on 1.30 min. The
widths of the peak bases were 0.45 min and 1.07 min. Calculate the following:

(a) Column resolution

𝟐[(𝒕𝑹 )𝑩 − (𝒕𝑹 )𝑨]

𝑹𝑺 =
𝑾𝑨 + 𝑾𝑩

= 2 (1.44-6.4) / (0.45+1.07) = 10.5

(b) The average number of plates in the column

N=16*(tR/W)2

For component A: NA = 16*(6.4/0.45)2 = 3.2 x 103

For component B: NB = 16*(14.4/1.07)2 = 2.9 x 103

N1 = (NA+ NB) / 2= (3.2 x 103 + 2.9 x 103) / 2 = 3.1 x 103 plates

 (c) The plate height

H=L/N

For component A:

HA=L/NA = (22.6 cm)/(3.2 x 103 plates) = 7.0 x 10-3 cm/plate

For component B:

HB = L/NB = (22.6 cm)/(2.9 x 103 plates) = 7.8 x 10-3 cm/plate

Hav= (HA +HB)/2 = ((7.8 + 7.0) x 10-3)/2 = 7.4 x 10-3 cm/plate

(d) The length of column required to achieve a resolution of 1.5

R1=10.5

N2=(RS2/RS1)2 x N1 = (1.5/10.5)2 x 3.1 x 103 = 63 plates

𝑁1 𝑅 2 𝑆1
=
𝑁2 𝑅 2 𝑆2

L=Hav x N2 = (7.4 x 10-3 cm/plate)(63 plates) = 0.46 cm

3. Suppose that you are to separate a mixture of benzoic acid, aspartame, and caffeine in a diet
soda by reverse phase HPLC. The following information is available to you.

tR in aqueous mobile phase buffered to a pH of

Compound
3.0 3.5 4.0 4.5

Benzoic acid 7.4 7.0 6.9 4.4

Aspartame 5.9 6.0 7.1 8.1

Caffeine 3.6 3.7 4.1 4.4

(a) Plot retention time versus pH for each compound on the same graph, and identify a pH
level that will yield an acceptable separation.
 pH versus tR
9
8
7
6
5
Benzoic acid
tR

4
3 Aspartame
2 Caffeine
1
0
0 1 2 3 4 5
pH

From the graph, the pH level that will yield an acceptable separation is pH=3.

(b) Calculate the selectivity for each pair of compounds at the optimal pH.

BA/ASP = 7.4/5.9 = 1.25 good separation

ASP/CAF = 5.93.69 = 1.6 good separation

4. A sample of the chromatogram of a mixture of acetaminophen, caffeine, salicylic acid and an

internal standard of benzoic acid run on a gas chromatograph is shown below.

(a) Calculate the column resolution for peak 1 and 2.

 Considering peaks 1 and 2, their retention times respectively are: 1.1 and 2.3
(tm=0.4), and their peak widths are (0.4 and 0.5):

The column resolution is calculated and

𝑹 = 𝟐. 𝟕

(b) What is the average number of plates in the column?

(𝑡𝑅 )1′ 1.1 − 0.4

𝑘1′ = = = 1.75
𝑡𝑚 0.4

(𝑡𝑅 )′2 2.3 − 0.4

𝑘2′ = = = 4.75
𝑡𝑚 0.4

1.75 + 4.75
𝑘𝑎𝑣𝑒 = = 3.25
2

(Note that you can take 2 adjacent peaks, i.e. peak 1 and 2 or peak 3 and 4. In this case I
take peak 1 and 2)

The selectivity (separation) factor is then calculated as:

𝑘2′ 4.75
𝛼= = = 2.71
𝑘1′ 1.75

The number of theoretical plates is calculated:

√𝑵  − 𝟏 𝒌𝟐
𝑹= ( )( )
𝟒  𝒌′𝒂𝒗𝒆 + 𝟏

 𝒌′𝒂𝒗𝒆 + 𝟏
𝑵 = 𝟏𝟔𝑹𝟐 ( )( )
−𝟏 𝒌𝟐

𝟐. 𝟕𝟏 𝟑. 𝟐𝟓 + 𝟏
𝑵 = 𝟏𝟔𝒙𝟐. 𝟕𝟐 ( )( ) = 𝟏𝟎𝟖. 𝟒
𝟐. 𝟕𝟏 − 𝟏 𝟒. 𝟕𝟓

(c) What is the plate height? The length of the column is given as 30 cm.

The plate height H=L/N= 30/ 108.4 = 0.277 cm