Synthetic Cannabinoid Receptor Agonists in Scottish Sub-Populations Thesis Submitted in Accordance with the Requirements of the University of Glasgow for the Degree of Doctor of Philosophy By Alice Seywright BSc (Hons) MSc MRSC Forensic Medicine and Science (School of Medicine, Dentistry and Nursing) March 2020 Acknowledgements Thanks, first and foremost, must go to my supervisors, Dr. Hazel Torrance and Dr. Fiona Wylie, for their help, guidance and support in conducting this research, obtaining samples and reviewing this thesis. The input from Denise McKeown was invaluable to the method development aspect of this work and my great thanks go also to her for this, and her general support. I would not have the opportunity to undertake this PhD were it not for Dr. Gail Cooper, who not only supported me through my MSc, but invited me back to the department of Forensic Medicine and Science to work and conduct this research. It is entirely possible I would have binned my work and given up entirely if it wasn’t for the fact that Lauren O’Connor was more often than not experiencing the same frustrations and challenges as I was. Thanks to Lauren for listening to my rants and for all her help and support throughout this… ordeal journey. Thanks also to Claire Parks for all her technical support, tending to QTrap1, and generally keeping me sane over the past few years. I genuinely would not have been able to complete this PhD without Fraser, most likely as I would have succumbed to hunger and rage. My dinner was always on the table when I got in; he has given hours of his life to proof reading and my practice presentations; and his overall support was immense. He certainly knows much more about SCRAs than the average Maths Teacher. Thank you. For general and miscellaneous advice and support, you cannot get much better than my dad, Ian, and mother-in-law Lindsay. They know where a semi-colon goes and exactly what word it is I know exists but cannot immediately place. My dad has also had the unenviable task of raising me in less than ideal circumstances and I owe him more than just thanks. No acknowledgements section is complete without mention of Tchai and Edinson. Their resigned acceptance of hugs, endured without struggle or scratching, got me through many a bad day of failed extractions and instrument breakdowns. i “Were there none who were discontented with what they have, the world would never reach anything better.” – Florence Nightingale “Life shrinks and expands in proportion to one’s courage” – Anaïs Nin ii Author’s Declaration “I declare that, except where explicit reference is made to the contribution of others, that this thesis is the result of my own work and has not been submitted for any other degree at the University of Glasgow or any other institution” Signature _______________________________ Printed name _______________________________ iii Table of Contents 1. Introduction ....................................................................................................... 1 1.1. Background ...................................................................................................... 1 1.2. Cannabinoid Receptors .................................................................................... 1 1.3. Phytocannabinoids ........................................................................................... 2 1.4. Endocannabinoids ............................................................................................ 3 1.5. Synthetic Cannabinoid Receptor Agonists ....................................................... 4 1.5.1. Chemistry and Nomenclature .................................................................. 14 1.5.2. Pharmacology .......................................................................................... 17 1.5.3. Prevalence and Risk of Harm .................................................................. 20 1.5.4. Analysis in Biological Matrices ................................................................. 23 2. Aims and Objectives ....................................................................................... 29 3. Development and Validation of a Method for the Detection and Quantitation of MDMB-CHMICA in Blood ...................................................................................... 30 3.1. Introduction ..................................................................................................... 30 3.2. Aims and Objectives ....................................................................................... 31 3.3. Materials ......................................................................................................... 31 3.3.1. Solutions .................................................................................................. 31 3.4. Method Development and Optimisation.......................................................... 34 3.4.1. Liquid Chromatography – Mass Spectrometry ......................................... 34 3.4.2. Extraction ................................................................................................. 35 3.4.3. Method Validation .................................................................................... 36 3.5. Results and Discussion .................................................................................. 38 3.5.1. Liquid-Chromatography – Mass Spectrometry ......................................... 38 3.5.2. Method Validation .................................................................................... 41 3.6. Conclusion ...................................................................................................... 50 4. Development and Validation of a Method for the Detection and Quantitation of Synthetic Cannabinoid Receptor Agonists in Blood and Urine ............................. 51 4.1. Aims and Objectives ....................................................................................... 51 iv 4.2. Materials ......................................................................................................... 51 4.3. Method ........................................................................................................... 52 4.3.1. Solutions .................................................................................................. 52 4.3.2. Selection of Analytes ............................................................................... 55 4.3.3. Liquid Chromatography – Mass Spectrometry ......................................... 56 4.3.4. Extraction of Analytes .............................................................................. 57 4.3.5. Method Validation – Blood ....................................................................... 62 4.3.6. Method Validation – Urine ........................................................................ 64 4.3.7. Validation – Intermediate Methods .......................................................... 65 4.3.8. Comparison of Prison ‘A’ and ‘B’ Samples ............................................... 66 4.3.9. Analysis of Drug Packets ......................................................................... 67 4.4. Results and Discussion .................................................................................. 70 4.4.1. Selection of Analytes ............................................................................... 70 4.4.2. Liquid Chromatography – Mass Spectrometry ......................................... 88 4.4.3. Extraction of Analytes ............................................................................ 103 4.4.4. Method Validation – Method 2.1 applied to blood .................................. 115 4.4.5. Method Validation – Methods 1.1, 1.2 and 2.1 applied to urine ............. 134 4.4.6. Comparison of Prison ‘A’ and ‘B’ Samples ............................................. 149 4.4.7. Analysis of Drug Packets ....................................................................... 152 4.5. Conclusion .................................................................................................... 153 5. Synthetic Cannabinoid Receptor Agonists in Scottish Sub-Populations ....... 155 5.1. Summary ...................................................................................................... 155 5.2. Individuals Undergoing Emergency Department Treatment ......................... 159 5.2.1. Introduction ............................................................................................ 159 5.2.2. Method ................................................................................................... 159 5.2.3. Results and Discussion.......................................................................... 160 5.2.4. Conclusion ............................................................................................. 177 5.3. Post-Mortem Casework ................................................................................ 178 5.3.1. Introduction ............................................................................................ 178 v 5.3.2. Method ................................................................................................... 178 5.3.3. Results and Discussion.......................................................................... 179 5.3.4. Conclusions ........................................................................................... 195 5.4. Individuals Admitted to or Liberated From a Scottish Prison Service Facility197 5.4.1. Introduction ...........................................................................................