June 2020 | Cannabis Prospect Magazine 3130 Cannabis Prospect Magazine | June 2020

MOLECULAR DIAGNOSTICS FOR THE HIGHEST QUALITY ASSURANCE IN CANNABIS PRODUCTS

Many of us seek alternative nutrition and medicine to deal with common ailments such as chronic pain, anxiety, infections, compromised immunity, etc. Although natural ingredients such as cannabis may successfully relieve

these issues, there are still concerns as to whether they are safe and effective. In fact, one of the most common consumer criticisms about cannabis products is that they do not receive a consistent experience when purchasing and consuming the same product at different times.

“The food, beverage and natural products industries have developed good manufacturing processes to ensure quality assur-ance in their supply chain. This is a common practice for brands so that they can consistently produce safe, quality products for consumers,”, explains Deleo de Leonardis, CEO at Purity-IQ.

SOLUTIONS FROM INNOVATIVE MOLECULAR SCIENCEScience provides solutions for quality assurance based on a foundation in genomics and metabolomics. The “omics” approach provides a key to understanding the considerable diversity of nat-ural chemicals, some of which induce a positive response amongst consumers, while others induce a negative effect. It is prudent that cannabis QA processes consider these omics-based molec-ular analyses that verify the production of safe, uniform, effective products.

The key concept in this molecular approach is found within the complexity of the genome as it relates to the metabolites produced by species that induce a specific physiological response in humans or even in our pets, as either nutrition or natural medicine. Can-nabis is a good example of a natural product we consume that is poorly understood as it has only recently been legalized in certain countries and allowed as an ingredient in edibles.

This is the first time the cannabis industry has had to deal with established brand owners, food manufacturers and retailers who require best practice and validated assurances for GMP and QA testing. There is an expectation that the cannabis-derived ingre-dients and products are fit for purpose, when it comes to safety, identity, purity and consistency. The challenge in designing a validated commercial identity test is that there are considerable knowledge gaps concerning cannabis cultivar (strain) characteriza-tion, given the genetic variation and mechanisms that control the

biochemical pathways producing agronomic traits and compounds known for their medicinal and psychoactive properties.

Although various studies have initiated cloning and expression of THC and CBD genes, variability in the cannabis genome has only been recently advanced to include multi-loci approaches for marker assisted breeding. These genomic advancements are important because they have identified extensive rearrangements at the THC/CBD acid synthase loci and new linkage groups asso-ciated with other important genes and gene networks, underlying cannabinoid and terpenoid accumulation. Such genome-wide association studies provide a basis for developing novel gene networks that are associated with traits including metabolites that can be used to characterize cultivars.

Our novel approach argues that current genomic sequencing alone is not enough to discriminate the complexity of cultivars. The basis for cultivar classification is facilitated by the considerable diversity of over 540 known compounds that have all been doc-umented in cannabis using analytical spectroscopy and chroma-tography focused on targeted phytochemicals. We developed a method that classifies chemical variation in cannabis cultivars using nuclear magnetic resonance (NMR) to analyze metabolites in an untargeted approach that explores the natural variation in cannabis phytochemistry.

This approach facilitates the detection of primary and second-ary metabolites including two-dimensional NMR spectrum signals that are directly proportionate to the molar concentrations of the chemical compounds. The method is easily and cost-effectively applied to commercial applications due to relatively simple sample preparation in non-destructive, deuterated solutions, which gener-ate immediate NMR spectra that are classified using multivariate statistical models we developed. This approach allows for the qual-ification of batch-to-batch variation in metabolite profiles based on confidence intervals.

Non-compliant batches can be rejected, which reduces the risk of contamination and ensures the consistency of cannabis prod-ucts. We call these modeled NMR profiles chemical fingerprints, which is a forensic quantitative measure of chemical diversity that can be correlated with several groups of traits. These include agronomic (e.g. growth, disease tolerance), quality (e.g. processing, stability and shelf-life), medicinal (e.g. THC & CBD and other can-

nabinoids such as Cannflavin), organoleptic sense-based cues including neurological, or experiential (e.g. taste, smell).

WHY ARE METABOLITE FINGERPRINTS USEFUL?

What is the benefit of having a cannabis metabolite fingerprint for the products that we consume? Metabolite fingerprints provide forensic evidence for any natural ingredient’s identity and purity allowing a manufacturer to produce a consistent product for consumers. This also provides a molecular diagnostic tool that filters un-natural synthetic adulterants that are health risks including improperly manufactured synthetic cannabinoids.

Metabolite fingerprints can help us understand how an individual will respond to a specific ingredient as we engage in large-scale human health trials in which many physiological responses are consid-ered including metabolite analyses of urine and blood. This is a personalized natural product experiential model in which con-sumers will be able to choose the cannabis cultivar that best supports their well-being. Consumers will have assurances that they are consuming the same cannabis product if it has the same metabolite fingerprint. Removing the mystery of how any natural products can improve your life is a com-mon goal in society at large and we seek to share science-based knowledge on how natural ingredients can help support a healthy lifestyle.

This is a process that is under the control of consumers who have the choice to demand 1) metabolite fingerprints from brand owners in order to ensure transpar-

ency in the supply of natural ingredients for the products they seek to purchase, and 2) clinical evidence to support health claims. This approach helps to assemble a science-based foundation for health prac-titioners so that they have the basic tools needed to recommend authentic cannabis cultivars with known metabolites for their patients who seek alternative medicine for chronic illnesses.

A FORENSIC APPROACH TO ENSURE QUALITY CANNABIS

The forensic world of nuclear magnetic res-onance that is used to eliminate counter-feiting of drugs can now be used to create metabolite fingerprints to ensure cannabis cultivar traceability and quality assurance. Our research team at the NHP Research Alliance, University of Guelph developed the concept of constructing NMR metabo-lomic fingerprint profiles to ensure identity and purity of specific cannabis cultivars for consumers (Figure 1 & 2). The proof of concept has been established using many commercial cannabis cultivars used in the marketplace and within wild populations throughout the globe.

“We are pleased to be working closely with the Guelph research team and Purity-IQ who are using a combina-tion of genomic sequencing and our NMR instrumentation for metabolomic analyses to assemble a detailed registry for cannabis cultivars that serves to advance cannabis science while providing a useful system that can verify cannabis cultivar identity and purity in the products for consumers,” stat-ed Henry Stronks, president at Bruker Ltd.

This facilitates testing of cannabis

Figures 1 & 2 Two samples of NMR metabolomic fingerprint profiles to ensure identity and purity of specific cannabis cultivars for consumers.

needed for GMP and verification of mate-rials in global trade. The cannabis cultivar registry is a database that can serve as an exploratory model for hypothesis testing on bioactivity of ingredients, efficacy-based human health trials, determination of toxic levels or dose, and agricultural produc-tion of commercial products. This “omics” database would also be a valuable food security tool for supply chain authentica-tion of ingredients through block-chain technology that can enable international trade of products of known quality, as they would be tagged with an inherent, forensic, metabolite fingerprint.

Based on history, every major manu-facturing sector has needed to implement rigorous ISO (International Standards Organization) based safety and quality standards to save lives, build quality and access markets, to effectively compete. Licenced cannabis producers of products derived from cannabis who wish to supply major retailers and established brand own-ers through contract manufacturing, will all eventually need to demonstrate their ability to consistently manage safety and quality assurances, which can be verified and validated by accredited third parties. Companies that can successfully integrate available industry quality tools into their day-to-day preventative and quality man-agement systems will be more competitive within a much broader marketplace.

Dr. Steven Newmaster, Director, NHP Research Alliance & Professor, University of Guelph

By Dr. Steven Newmaster, Director, NHP Research Alliance & Professor, University of Guelph