HPLC

High Performance Liquid Chromatography

WJEC IVQ Science for Work Unit 3 WJEC Pathways Science for Work Unit 2.8 WJEC GCSE Additional Applied Science

Diagram HPLCsolvent

reservoir

pump

detector

waste

injection port

HPLCSyringe

HPLC column

This is where separation occurs

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 listCaffeine• Caffeine is also used athletes as a performance aid• The IOC presently lists caffeine as a banned substanceAmphetamine• These may mean an athlete feels less tied and can endure longer• All forms of amphetamines are banned by the IOCUrea• 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

Temperature: 25 oCMobile phase: 50% water

50% acetonitrileFlow rate: 2 cm3 /minColumn: C-18

Analyst: Ann Lyst

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Aspirin Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Caffeine Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Amphetamine Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Urea Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Athlete A Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Athlete B Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Athlete C Return to menu

0 1.0 2.01.5 2.5 3.0 4.03.5Time (minutes)

0.5

Athlete A and overlay Return to menu

aspirincaffeineurea

amphetamine