Index of /rozprawy2/10907

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List of contents

1.1.1. Definition and background ... 11

1.1.2. Cytochromes P450 superfamily ... 13

1.1.3. Strategies of metabolism investigation ... 15

1.2. Drugs of abuse ... 18

1.2.1. What is addiction ... 18

1.2.2. Main groups of narcotics ... 18

1.2.3. Exemplary drugs metabolism pathways ... 21

1.3. Analytical techniques used in drug metabolism study ... 23

1.3.1. Electrochemistry ... 23

1.3.2. Mass spectrometry ... 25

2. Aim of the study ... 28

3. Publications ... 29

4. Materials and methods ... 30

4.1. Chemicals and reagents ... 30

4.2. Instrumentation ... 31

4.3. Experiments with cocaine ... 34

4.3.1. Electrochemical system connected on-line with mass spectrometry (EC/MS) ... 34

4.3.2. Off-line EC/LC-MSn analysis ... 36

4.3.3. Maintenance of working electrodes ... 37

4.3.4. Liquid chromatography coupled with mass spectrometry detection (LC-MSn) ... 37

4.3.5. Preparation of the rat liver microsomes ... 38

4.3.6. Cocaine metabolites, preparation with application of liver microsomes .. 39

4.4. Electrochemical oxidation of the selected psychoactive substances ... 39

4.4.1. Conditions of the experiment ... 39

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4.4.3. Electrochemical oxidation of psychoactive compounds with FAPA-MS

detection ... 41

4.4.4. Electrochemical oxidation of selected psychoactive compounds with ESI-MS detection ... 42

4.5. Simulation of selegiline metabolism ... 42

4.5.1. Electrochemical oxidation of selegiline on GC and BDD electrodes ... 42

4.5.2. Generation of selegiline metabolites using rat and human liver microsomes ... 43

4.5.3. EC/LC/ESI-MS analysis ... 44

4.5.4. EC/LC/DBDI-MS analysis ... 45

5. Results and discussion ... 46

5.1. Results of the experiments with cocaine ... 46

5.1.1. Electrochemical oxidation of cocaine on BDD working electrode with on-line mass spectrometry detection ... 46

5.1.2. Off-line EC/LC-MSn analysis ... 51

5.1.3. Cocaine oxidation on GC working electrode ... 53

5.1.4. Cocaine metabolites obtained by incubation with rat microsomes ... 54

5.2. Electrochemical oxidation of selected psychoactive substances ... 55

5.2.1. Coupling EC with FAPA-MS detection ... 55

5.2.2. FAPA-MS identification of oxidation products ... 55

5.2.3. ESI-MS identification of oxidation products ... 59

5.3. Simulation of selegiline metabolism ... 69

5.3.1. Electrochemical oxidation of selegiline ... 69

5.3.2. Generation of selegiline metabolites by incubation with microsomes ... 69

5.3.3. Electrochemical oxidation of selegiline ... 73

6. Final remarks ... 79

7. Conclusions ... 83

Streszczenie ... 85