1. HPLC High Performance Liquid Chromatography WJEC Pathways Applied Science Unit 2.8 WJEC GCSE Additional Applied Science

2. Diagram HPLC solvent reservoir pump detector waste injection port HPLC Syringe HPLC column This is where separation occurs

4. Testing for drugs HPLC can be used to test for ‘performance enhancing’ drugs that may be used by athletes

5. 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

6. 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

7. Choose the sample to run Aspirin Caffeine amphetamine Urea Athlete A Athlete B Athlete C Athlete A -overlay Conditions of analysis Temperature:25 o C Mobile phase: 50% water 50% acetonitrile Flow rate:2 cm 3 /min Column:C-18 Analyst:Ann Lyst

8. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 AspirinReturn to menu

9. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 CaffeineReturn to menu

10. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 AmphetamineReturn to menu

11. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 UreaReturn to menu

12. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 Athlete AReturn to menu

13. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 Athlete BReturn to menu

14. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 Athlete CReturn to menu

15. 01.02.01.52.53.04.03.5 Time (minutes) 0.5 Athlete A and overlayReturn to menu aspirin caffeine urea amphetamine