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KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY (KNUST) COLLEGE OF SCIENCES FACULTY OF BIOSCIENCE DEPARTMENT OF BIOCHEMISTRY AND BIOTECHNOLOGY EVIDENCE OF ALCOHOLIC BEVERAGES LACED WITH CANNABIS SOLD IN BARS IN THE ACCRA METROPOLIS THIS THESIS IS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF AN MSc IN FORENSIC SCIENCE KAFUI NKROMA APRIL, 2018 i DECLARATION I hereby declare that this submission is my own work towards the MSc. And that, to the best of my knowledge; it contains no material previously published by another person or material which has been accepted for the award of any other degree of the University except where due acknowledgement has been made in the text. Kafui Nkroma ………………………… ………………………… (PG4429115) Signature Date Certified By: Dr Samson Pandam Salifu ………………………… ………………………… (Supervisor) Signature Date Dr Peter Twumasi ………………………… ………………………… (Head of Department) Signature Date ii ACKNOWLEDGEMENT Firstly, am most grateful to the almighty God whose strength and inspiration has seen me through this study. I wish to express my sincerest gratitude to my supervisor Dr Pandam Samson Salifu, and Dr Christpher Larbie for their patience, guidance and constructive criticism which made it possible for the completion of this thesis. I thank the Head and entire staff of the Department of Biochemistry and Biotechnology of KNUST and Forensic tutors for their encouragement. My special gratitude goes to Mr James Ataki, and Dr F.C.Mills-Robertson who took part in making this endeavor possible. I would like to acknowledge Dr Paul Fosu and Mr Peter Ashittey and the entire staff of Standards Authority for granting me access to their facilities to enable me complete my thesis. My final thanks goes to Mr Michael Akleshie for his contributions in diverse ways to the success of this work. iii ABSTRACT Alcohol is consumed for recreational purposes and it is not a strictly controlled substance. Misuse of alcohol can lead to severe consequences – socially and health of an individual. Alcohol is sold in bars, pubs and recreational places where people meet to chat, share a drink and to socialize. The aim of this work was to investigate potential adulteration of cannabis in alcoholic beverages that are sold in bars in the Accra Metropolis. Sixty (60) local and foreign alcoholic beverages were sampled from bars and pubs in the slums, unplanned and planned areas in the Accra Metropolis. The samples were first screened for presence of cannabis using presumptive Fast Blue B test and GC-MS was used for the identification, and quantification of the three main cannabinoids; tetrahydrocannabinol (Δ9 – THC), cannabinol (CBN) and cannabidiol (CBD) in the alcoholic beverages. The locally produced alcoholic beverages had the highest level of CBN (193.87%) while the foreign alcoholic beverage sampled from another location of Accra had highest value of 96.21% (w/w) Δ9 – THC. The average value of CBN (25.93%) was greater than the average Δ9 – THC content (5.25%). This means the cannabis used as adulterants were of old stock. This work uncovered higher levels of cannabis in alcoholic beverages sold in the Accra Metropolis. This could have health implications to customers who unknowingly purchased and consumed alcoholic beverages adulterated with cannabis that are sold in local bars in the Accra Metropolis. To the authors knowledge, this work is first of its kind to demonstrate lacing of alcoholic beverages with hazardous cannabis sold in bars. iv TABLE OF CONTENTS DECLARATION………………………………………………………………… i ACKNOWLEDGEMENT…………………………………………..................... ii ABSTRACT……………………………………………………...……….……… iii TABLE OF CONTENTS…………………………………………………........... iv LIST OF TABLES………………………………………………………….......... v LIST OF FIGURES……………………………………………………………… vi CHAPTER ONE…………………………………………….…………………… 1 1.0 INTRODUCTION……………………………………………………… 1 1.1 Problem Statement…………………………………………………… 3 1.2 Main objective………………………………………………………... 3 1.2.1Specific objectives……………………………………………………. 3 1.3 Justification…………………………………………………………….. 4 CHAPTER TWO…………………………………………………………………. 5 2.0 LITERATURE REVIEW……………………………………………… 5 2.1 Psychoactive substances……………………………………………... 5 2.2 Cannabis……………………………………………………………… 7 2.2.1 Cannabis Strains………………………………………………………9 2.2.2 Hybrid varieties of cannabis……………………………………….....9 2.2.2.1 Hemp………………………………………………………………..9 2.2.2.2 White widow………………………………………………………..10 2.2..2.3 Skunk…………………………………………………………….....10 2.2.2.4Sinsemilla…………………………………………………………….10 v 2.2.3 Forms of Cannabis……………………………………………………10 2.2.3.1 Herbal Cannabis……………………………………………………11 2.2.3.2 Hashish……………………………………………………………...11 2.2.3.3 Hashish oil…………………………………………………….........12 2..3 Cannabinnoids ………..……………………………………………….13 2.3.1 Tetrahydrocannabinol…………………………………………………14 2.3.2 Cannabidiol……………………………………………………………15 2.3.3 Cannabinol…………………………………………………………….16 2.3.4 Other Cannabinnoids………………………………………………….17 2.3.5 Cannabinnoid receptor 1…………………………………………….. 18 2.3.6 Cannabinnoid receptor 2……..……………………………………….18 2.3.7Cannabis edibles…..…………………………………………………..18 2.3.8 Effects of Cannabis use……………………………………………… 19 2.4 Cannabis Dependence………………………………………………… 19 2.5 Cannabis Potency Trends……………………………………………...22 2.5.1 Potency testing in Cannabis…………………………………………..23 2.5.2 The function of Precursors in Potency testing……………………….23 2.6 The chemistry of alcoholic beverages……………………………….. 24 2.6.1 Alcoholic beverages…………………………………………………. 25 2.6.2 Distilled and alcoholic undistilled beverages……………………….. 26 2.6.3 Undistilled alcoholic beverages………………………………………26 2.6.4 Distilled alcoholic beverages…………………………………………26 2.6.5 Gin Production………………………………………………………..27 2.6.6 Pot distilled gin………………………………………………………..28 2.6.7 Column distilled gin………………………………………………… 28 vi 2.6.8 Compound gin………………………………………………………. 28 2.6 .9Whiskey Production………………………………………………….28 2.7. Akpeteshie Production………………………………………………...29 2.7.1Long term and short term effects of alcohol………………………….30 2.7.1Breath analysers and blood alcohol content…………………………. 30 2.7.2 Alcohol dependence………………………………………………… 31 2.7.3 Treatment of alcohol dependent people…………………………….. 32 2.8 Instrumental methods……………………………………………….. 33 2.8.1 Gas chromatographic mass spectrometer (GC-MS)………………… 33 2.8.2 The Column…………………………………………………………. 34 2.8.3 Carrier gases………………………………………………………… 35 2.8.4 Injection system…………………………………………………….. 35 2.8.5 The detector………………………………………………………….. 36 2.8.6 Sample ionization…………………………………………………….. 36 2.8.7 Electron ionization…………………………………………………… 36 2.8.8 Cold ionization………………………………………………………. 36 2.8.9 Chemical ionization…………………………………………………. 37 2.9 Analysis on the GC-MS………………………………………………. 37 2.9.1 Rural Urban Migration………………………………………………. 37 2.9.2 Slums………………………………………………………………… 38 2.9.3 Unplanned Areas…………………………………………………….. 38 2.9.4 Planned areas………………………………………………………… 38 CHAPTER THREE………………………………………………………………. 40 3.0 MATERIALS AND METHODS……………………………………… 40 3.1 Study area……………………………………………………………… 40 vii 3.2 Study design…………………………………………………………… 41 3.3 Sample collection………………………………………………………..41 3.4 Materials……………………………………………………………………...42 3.4.1 Fast Blue B salt test…………………………………………………………42 3.5 Confirmatory analysis………………………………………………………..43 3.5.1 GC-MS Analysis…………………………………………………………...43 3.5.2 Chemicals and standards used for analysis………………………………..43 3.5.3 Sample preparation………………………………………………………...44 3.5.4 Gas Chromatographic conditions………………………………………….44 3.5.5 Analysis of samples on the GC-MS………………………………………..44 3.6 Statistical Data…………………… ………………………………………….45 CHAPTER FOUR………………………………………………………………… 46 4.0 RESULTS…………………………………………………………………... 46 4.1 Introduction……………………………………………………………….. 46 4.2 Presumptive test result for local and foreign alcoholic beverages………… 46 4.3 Determination of CBN Δ9 – THC and CBD with the GC-MS…………… 47 4.3.1 Cannabinnoid concentrations in the Slum areas………………………….. 54 4.3.2 Cannabinnoid concentrations in Planned areas…………………………… 56 4.3.3 Cannabinnoid concentrations in the Unplanned Areas…………………… 57 4.3.4 Comparism Mean of Δ9–THC, CBN and CBD concentration in alcoholic beverages………………………………………………………………….. 59 CHAPTER FIVE.……………………………………………………………… 60 5.0 DISCUSSION…...………………………………………………………… 60 5.1 Introduction………………………………………………………………… 60 CHAPTER SIX..……..………….…………………………………………….. 61 viii 6.0 CONCLUSION AND RECCOMMENDATION…………………………... 61 6.1 Conclusion…………………………………………………………………. 61 6.2 Recommendation…………………………………………………………… 65 REFERENCES………………………………………………………………… 66 APPENDIX 1…………………………………………………………………… 79 ix LIST OF TABLES 4.1 Canabinnoid concentration of alcoholic beverages in the Slums…………….....54 4.2 Canabinnoid concentration of alcoholic beverages in the Planned Areas…........55 4.3 Canabinnoid concentration of alcoholic beverages in the Unplanned Areas…...58 4.4 Comparison of Means using independent two sample t-test..…………………..59 x LIST OF FIGURES 2.1 An illustration of the major elements of neurotransmission…………………… 6 2.2 The three main strains of cannabis……………………………………………..9 2.3 Herbal cannabis…………………………………………………………………11 2.4 Hashish………………………………………………………………………….12 2.5 Hashish oil……………………………………………………………………...13 2.6 Chemical Structure of Tetrahydrocannabinnol (Δ9-THC)…..………………..14 2.7 Decarboxylation of Δ9-THC acid to Δ9-THC…………………………..........15 2.8 Chemical Structure of Cannabidiol……………………………………………16 2.9 Chemical Structure of Cannabinol…………………………………………....17 2.10 Chemical Structure of Alcohol…………………………………………….....24 2.11 Local Production of Akpeteshie……………………………………………..29 2.12 Parts of a Gas Chromatography Spectrophotometer………………………...34 3.1