UNIVERSITI KUALA LUMPUR

MALAYSIAN INSTITUTE OF CHEMICAL & BIOENGINEERING

TECHNOLOGY

TUTORIAL 6 : High Performance Liquid Chromatography (HPLC)

1. Explain about High Performance Liquid Chromatography (HPLC)?

2. Give and describe a function of the basic component in HPLC?
3. Define the following term: Mobile phase, Stationary phase, Reverse phase, Normal
phase.
4. Explain how the separation of a mixture can be occurs in HPLC.
5. Identify four types High Performance Liquid Chromatography (HPLC).
6. Name and describe two types of partition chromatography which operate based upon
the relative polarities of the mobile phase and stationary phases.
7. Explain about how separation occur in size exclusion chromatography.
8. Identify a factor that may reduce the accuracy of result obtain by using HPLC.
9. Name and explain two types of mobile phases common apply in HPLC.
10. You are request to determine the amount Benzoic acid in a soft drink using High
Performance Liquid Chromatography (HPLC). You are given a stock of benzoic acid
standard 1000 ppm, and are request to prepare a series of dilution of 50 ppm, 100 ppm,
150 ppm, 200 ppm in 50 ml volumetric flask.
(i) Draw flowchart for such analysis
(ii) Show all calculation for preparation of series of dilution
(iii) Roughly sketch an expected standard calibration curve for your standard.
(iv) Show how to determine amount of caffeine in a cup of coffee by hand from your
standard calibration curve. (Roughly Sketch).
11. Briefly explain about Preparative HPLC and LC–MS.
12. What is the main advantage of HPLC compare to GC?
13. Name typical applications of ion exchange chromatography.
14. List down application of High Performance Liquid Chromatography (HPLC)