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Chromatography Tutorial Questions and Analysis

This document is a tutorial for CHM 260 Basic Instrumental Analysis focusing on chromatography. It contains a series of questions and calculations related to various chromatography techniques, including ion-exchange and size exclusion chromatography, capacity factors, selectivity factors, and resolutions between components. The tutorial also addresses the effects of different conditions on plate height and provides practical scenarios for analysis using retention times and peak widths.

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Teratak Biru
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48 views2 pages

Chromatography Tutorial Questions and Analysis

This document is a tutorial for CHM 260 Basic Instrumental Analysis focusing on chromatography. It contains a series of questions and calculations related to various chromatography techniques, including ion-exchange and size exclusion chromatography, capacity factors, selectivity factors, and resolutions between components. The tutorial also addresses the effects of different conditions on plate height and provides practical scenarios for analysis using retention times and peak widths.

Uploaded by

Teratak Biru
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd

ACADEMIC SESSION 2016/2017 SESSION 1: JUN – OCTOBER 2016

CHM 260 BASIC INSTRUMENTAL ANALYSIS


TUTORIAL 4: AN INTRODUCTION TO CHROMATOGRAPHY
PAHANG BRANCH SEPARATIONS [Type text]

Name: Student No:

Answer the following questions.

1) Describe the fundamental difference between ion-exchange and size exclusion chromatography.

2) Given the following data:

Compound Retention time (min)


Unretained species 0.87
Methyl paraben 3.23
Propyl paraben 5.16

a) Calculate the capacity factor, k’ for methyl paraben and propyl paraben.
b) Based on the k’ value obtained in (a), comment on the elution rate of methyl paraben.
c) Calculate the selectivity factor, α for the two-component mixture.

3) In a high performance liquid chromatography (HPLC) analysis, toluene exhibits a capacity factor
of 3.50 on a 15.0-cm column. The unretained solute is eluted in 1.33 min.

a) Find the retention time, tR of toluene.


b) Calculate how many theoretical plates would produce a toluene peak eluting at 12.83 min
with a width at the base of 18.4 s.
c) Determine the plate height of the peak in (b).

4) A chromatography experiment was done by injecting a mixture of two components, X and Y onto
a chromatographic column with 2.00 x 10 3 theoretical plates (N). The following results were
obtained.

Parameter Dead Time, tM Component X Component Y


Capacity factor, k’ - 3.50 5.45
Retention time, tR 3.33 min 15.00 min 21.5 min

Calculate:
a) Width at the base, Wb for each peak.
b) The resolution, Rs between components X and Y.

5) State the effect of the following conditions on the plate height of a chromatographic column.
Indicate whether the plate height will increase or decrease.

a) Increasing the flow rate of the mobile phase.


b) Increasing the diameter of the capillary column.

S.N. Jaafar 1
ACADEMIC SESSION 2016/2017 SESSION 1: JUN – OCTOBER 2016
CHM 260 BASIC INSTRUMENTAL ANALYSIS
TUTORIAL 4: AN INTRODUCTION TO CHROMATOGRAPHY
PAHANG BRANCH SEPARATIONS [Type text]
6) The separation of compound X, Y and Z on a chromatographic column is shown in the following
table:

Compound Retention time (min) Peak width at base (min)


X 2.5 0.15
Y 3.8 0.30
Z 4.5 0.55

The time taken for the unretained species was 0.5 min.
a) Calculate the capacity factor (k’) for compound X and compound Z.
b) Determine the resolution (Rs) of compound Y and Z.

7) The chromatograms of compounds A and B were obtained at the same flow rate using two
columns, Column 1 and Column 2, of equal length.

Choose:
a) Which column (1 or 2) has a higher H?
b) Which compound (A or B) has a higher affinity towards the mobile phase?

8) Methanol and ethanol are separated using gas chromatography (GC) giving retention times of
370 s and 385 s, respectively. The base width of each peaks are 16.0 s and 17.0 s, respectively.
An unretained air peak occurs at 10.0 s.
a) Calculate the capacity factor, α , for this separation.
b) Calculate the resolution, Rs, between the methanol and ethanol peaks. Are the methanol
and ethanol peaks completely resolved?

9) What observation on a chromatogram would lead you to conclude that too much sample have
been injected into the chromatographic column?

S.N. Jaafar 2

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