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Tutorial 6 CHM260

The document is a tutorial for CHM 260 Basic Instrumental Analysis focusing on High Performance Liquid Chromatography (HPLC) for the academic session 2016/2017. It includes questions on characteristics of mobile phases, types of chromatography, elution methods, and practical applications in HPLC analysis. The tutorial aims to assess students' understanding of HPLC principles and techniques through various analytical scenarios.

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Teratak Biru
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20 views2 pages

Tutorial 6 CHM260

The document is a tutorial for CHM 260 Basic Instrumental Analysis focusing on High Performance Liquid Chromatography (HPLC) for the academic session 2016/2017. It includes questions on characteristics of mobile phases, types of chromatography, elution methods, and practical applications in HPLC analysis. The tutorial aims to assess students' understanding of HPLC principles and techniques through various analytical scenarios.

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

CHM 260 BASIC INSTRUMENTAL ANALYSIS


TUTORIAL 6: HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
PAHANG BRANCH [Type text]

Name: Student No:

Answer the following questions.

1) Identify three (3) important characteristics of a good mobile phase for HPLC.

2) A mixture consisting of compound A, B and C is separated on a C 18 column with a mobile phase


of 20:80 % (v/v) ethanol:water,

a) Is this a normal phase or reversed phase chromatography? Explain briefly your answer.
b) Given the polarity of solutes: A>B>C. Rank the elution order of the compounds from the
lowest to highest retention time.
c) If a 50:50 % (v/v) solution of ethanol and water is used instead, predict how the retention
times of the compounds will change (the same, longer or shorter) compared to the first
run.

3) Differentiate between a normal-phase and reversed-phase HPLC based on the polarity of mobile
phase and the stationary phase used.

4) Relate how degassing of the mobile phase can affect the performance of a HPLC detector.

5) In HPLC,

a) Justify why gradient elution is the preferred method over isocratic elution.
b) Differentiate between isocratic and gradient elution.

6) Given below is the chromatogram of a sample mixture containing compounds A, B and C which
been analyzed using HPLC normal-phase column. Determine the sequence of compounds in the
order of increasing polarity.

S.N. Jaafar 1
ACADEMIC SESSION 2016/2017 SESSION 1: JUN – OCTOBER 2016
CHM 260 BASIC INSTRUMENTAL ANALYSIS
TUTORIAL 6: HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
PAHANG BRANCH [Type text]
7) Suggest a stationary phase-mobile phase combination of a normal phase HPLC.

8) A student is running an HPLC sugar analysis using a C 18 bonded column in a 1:4 water:methanol
solvent system. He found two peaks in his chromatogram. D-fructose (labeled as 1) and D-
glucose (labeled as 2).

a) Based on the chromatogram, conclude which sugar is more polar.


b) Suggest a method to decrease the elution time of these sugars.
c) Justify your answer in (b).

9) In HPLC, distinguish between the function of a guard column and a He sparger.

10) You are required to separate a mixture of polymers by collecting the highest molecular weight
compound as the first fraction. Choose the most suitable liquid chromatography technique you
would use to accomplish this task.

S.N. Jaafar 2

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