Synthetic cannabinoid method development with Saccharomyces cerevisiae and Escherichia coli proteome patterns

Olipoh, George

http://hdl.handle.net/10400.26/19074

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Authors

Olipoh, George

Advisor(s)

Quintas, Alexandre

Costa, Isabel Margarida

Abstract(s)

The emergence and abuse of new synthetic cannabinoids has been on the rise because legal regulations against particular derivatives of cannabinoids tend to cause the introduction of new variants as the law is made for specific molecules and not for a group of molecules. They come with no clinical indications of use on their labels, and vaguely described intoxicating effects hence they were not banned. Consequently, they are also referred to as ‘Smart Drugs’ or more formally ‘New Psychoactive Substances’ (NPSS). Many classified synthetic cannabinoid agonists remain available to users in many countries mainly through ‘smart’ and online shops. The challenges posed to the regulation of these products are due to their ever-changing composition, adulterants and variable vehicles of distribution. Synthetic cannabinoids are famous for their recreational effects similar to that of marijuana, which has neuropsychiatric effects due to the action of its primary active ingredient, Δ9-tetrahydrocannabinol (Δ9-THC), binding to endogenous cannabinoid receptors. This study was conducted first to find the best microorganism model for the development of a new screening method for the analysis of synthetic cannabinoids by using the proteome expression patterns obtained from proteins of Saccharomyces cerevisiae and E. coli. The research also sought to find an efficient protein extraction and concentration method for expressed microorganism proteins for two-dimension gel electrophoresis. Saccharomyces cerevisiae and Escherichia coli were studied in this work. S. cerevisiae rather than E. coli proved to be a better microorganism model for to be exploited for findings in this research. Diclofenac and JWH-018 standard were used as test drugs. The Amicon® membrane filter concentration method was found to be the most efficient with protein yield greater than 20mg/mL for 500mL cell culture volume. Even though twodimension gels did not show a large number of protein spots, the application of more concentrated protein solution from S. Cerevisiae is paramount to get a better proteomic pattern for use as a sensor.