HPLC - WJEC

Report
HPLC
High Performance Liquid
Chromatography
WJEC Pathways Applied Science Unit 2.8
WJEC GCSE Additional Applied Science
Diagram HPLC
solvent
reservoir
pump
This is where
separation occurs
detector
HPLC
Syringe
injection
port
HPLC
column
waste
Testing for drugs
• HPLC can be used to test for ‘performance
enhancing’ drugs that may be used by athletes
What are we looking for?
Aspirin
• Some endurance athletes have been drugs such as aspirin before and
during competition in an attempt to compete at the highest intensity for
the longest duration
• Aspirin is also taken to relieve symptoms of colds etc
• Not on the International Olympic Committee (IOC) banned list
Caffeine
• Caffeine is also used athletes as a performance aid
• The IOC presently lists caffeine as a banned substance
Amphetamine
• These may mean an athlete feels less tied and can endure longer
• All forms of amphetamines are banned by the IOC
Urea
• Naturally formed in the body
What is in the sample?
Who has broken the rules?
• Identify what is present in each sample of
blood taken from three different athletes.
• All samples were taken under the same
conditions
Choose the sample to run
•
•
•
•
•
•
•
•
Aspirin
Caffeine
amphetamine
Urea
Athlete A
Athlete B
Athlete C
Athlete A -overlay
Conditions of analysis
Flow rate:
Column:
25 oC
50% water
50% acetonitrile
2 cm3 /min
C-18
Analyst:
Ann Lyst
Temperature:
Mobile phase:
Aspirin
0
0.5
1.0
1.5
2.0
Return to menu
2.5
3.0
3.5
4.0
Time (minutes)
Caffeine
0
0.5
1.0
1.5
2.0
Return to menu
2.5
3.0
3.5
4.0
Time (minutes)
Amphetamine
0
0.5
1.0
1.5
2.0
2.5
Return to menu
3.0
3.5
4.0
Time (minutes)
Urea
0
0.5
1.0
1.5
2.0
Return to menu
2.5
3.0
3.5
4.0
Time (minutes)
Athlete A
0
0.5
1.0
1.5
2.0
Return to menu
2.5
3.0
3.5
4.0
Time (minutes)
Athlete B
0
0.5
1.0
1.5
2.0
Return to menu
2.5
3.0
3.5
4.0
Time (minutes)
Athlete C
0
0.5
1.0
1.5
2.0
Return to menu
2.5
3.0
3.5
4.0
Time (minutes)
aspirin
caffeine
urea
amphetamine
0
0.5
1.0
Athlete A and overlay
1.5
2.0
2.5
3.0
Return to menu
3.5
4.0
Time (minutes)

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