Cannabis: Trends in Analytical Research

Introduction

2018 was a big year for cannabis. Vermont became the first state to legalize recreational cannabis by state legislature, a growing number of lawmakers (including those intending to run in the 2020 presidential election) voiced their support for legalizing cannabis, as did voters in a number of midwestern and red-voting states, and the US Food and Drug Administration (FDA) approved a drug derived from marijuana for the first time. Meanwhile, north of the border, Canada became the second country in the world to legalize the drug. We are currently at a critical point in the creeping legalization of the drug in both recreational and medical settings.

However, even with these developments and in spite of demand from patient groups, clinical research into the drug is still hindered by federal laws and the imperative to supply consumers with a safe and consistent product is stymied by a lack of standardization at multiple stages from “seed-to-sale”.

The cannabis analysis program at Pittcon 2019 will open with a talk from Joshua Crossney of Jcanna, a non-profit focused on medicinal cannabis and the advancement of cannabis analytical testing technology.

Crossney will argue that in order to improve the quality of medical cannabis we need to “bridge the gap” between analytical sciences and the medical cannabis industry. He will describe recent activities that have taken place to educate and bring together people from disparate fields with a shared interest in this area. This includes, article publication, conferences, networking sessions and cross-disciplinary workshop sessions.

Crossney will also consider the forces that are currently either hindering or advancing the fields of cannabis testing and medical cannabis. Topics covered include the current medical curricula, federal laws that prevent cannabis research and a lack of standardization in testing regulations, analytical methods between labs and laws between states.

This year’s Pittcon takes place in Philadelphia, Pennsylvania March 17-21, 2019. Cannabis was a major topic at Pittcon 2018 and will feature highly again on this year’s programme, reflecting that interest in this area is far from diminishing. At Pittcon 2019, the focus will turn to the challenges involved in clinical research and cannabis testing, and the role advances in analytical methods are playing in these. There are three cannabis symposia at this year’s conference: Analytical Cannabis I, Analytical Cannabis II, and Overcoming Challenges and Streamlining Cannabis Containment Testing. They will feature leading experts from fields as diverse as clinical research, chemistry, systems engineering, and patient advocacy.

Not only will there be a range of talks on the latest developments in this sector but at the Pittcon 2019 expo, you can meet representatives from all the major companies delivering technology in the cannabis analysis space. These include Shimadzu, who have brought to market a dedicated cannabis analyzer, Bruker, Phenomenex and Merck (previously known as Millipore Sigma).

There is no better time to get up to date with the latest developments in the field of cannabis analytical research, and no better place to do it than at this year’s Pittcon.

References

• Wikipedia. Timeline of cannabis laws in the United States. Available at: https://en.wikipedia.org/wiki/Timeline_of_cannabis_laws_in_the_United_States#cite_note-56. Accessed: March 2019.
• Forbes (2018) Marijuana’s 10 biggest victories of 2018. Available at: https://www.forbes.com/sites/tomangell/2018/12/06/marijuanas-ten-biggest-victories-of-2018/#1307bac132df Accessed: March 2019.
• Food & Drug Administration (2018). FDA approves first drug comprised of an active ingredient derived from marijuana to treat rare, severe forms of epilepsy. Available at: https://www.fda.gov/newsevents/newsroom/pressannouncements/ucm611046.htm Accessed March 2019.

Chapter 1 – Current Challenges for Cannabis Research

We are seeing the modern medical cannabis industry at its very beginnings. Interest in this area is growing among researchers, clinicians and patients but this has not been enough to overcome many of the barriers that come with studying a drug that has been illegal for almost 100 years. Researchers find themselves caught in a catch-22: clinician’s do not feel confident prescribing the drug because of a lack of gold-standard evidence to support it, but that evidence cannot be gathered because research is impeded at multiple steps. In the USA, where many states have legalized medicinal or recreational marijuana, federal legislation stalls researchers’ attempts to study the clinical effects of cannabis. Additionally, there are influential groups who are opposed to the efficacy of cannabis being studied in randomized controlled trials, including some in Big Pharma, politics and law enforcement.

In recent years there has been a clear shift in public attitudes to marijuana within the western world and in the USA this has been proven at the ballot box where voters in multiple states have expressed support for legalizing medical or recreational use of the drug.

In the USA, research institutions must pass through multiple hoops if they wish to carry out research on cannabis. This includes sourcing their cannabis from an institution approved by the National Institute on Drug Abuse (NIDA), of which there is only one – the University of Mississippi. Some researchers have complained that the nature of cannabis available via NIDA are not representative of those being used recreationally or medicinally in the many states that have now approved medicinal use of the herb. These differences include the strength of cannabis available, with retail products being much more potent than those available for study, different storage conditions, such as freezing, and different presentations, such as oils, waxes and edibles.

While NIDA does finance some research into medicinal cannabis, this is not actually part of its modus operandi, meaning the vast majority of its research budget goes towards exploring the harms of cannabis and prevention of these.

While there is both anecdotal evidence and observational studies that indicate therapeutic benefits of cannabis in certain medical conditions, these are not a replacement for properly controled randomized trials. A case in point: the UK approved legislation in November 2018 to allow the provision of medical cannabis to named individuals, but so far prescriptions have been unforthcoming. Reports suggest that this is due to risk-averse clinicians’ reluctance to prescribe a drug effectively off-label without sufficient evidence from clinical trials.

At Pittcon 2019, we will hear from Sue Sisley of the Scottsdale Research Institute, in Phoenix, Arizona. Sisley has been at the forefront of efforts to conduct clinical research into natural cannabis flower. Her research institute received the first FDA-approval to conduct a clinical trial into the efficacy of marijuana in the treatment of veterans with post-traumatic stress disorder. However, despite this approval being granted in 2011, the trial has yet to begin. In her Pittcon presentation, Sisley will discuss the obstacles that she and her team have faced trying to get cannabis flower through the FDA drug development process.

As an increasing number of US states make legal provision for recreational and medical marijuana, so too does the demand for methods for producers to provide high-quality, consistent products and ultimately, to operate competitively within a growing market.

At Pittcon 2019, Markus Roggen, a specialist in extraction from the Vancouver-based Complex Biotech Discovery Ventures, will explain how the particular expansion in the market for cannabis concentrate and infused products has created a need for improved extraction techniques. His talk will explore how pre- and post-extraction steps effect efficiency, and the quality and quantity of the outcome product. Factors involved that are particularly relevant include mean particle size, particle size distribution and packing density in the extractor column.

Outside of the symposia at Pittcon this year, Fritsch, who offer sample-preparation tools such as the Pulverisette 14, which provides homogenous particle size reduction of cannabis samples with no loss of cannabinoids or terpenes.

References

• Multidisciplinary Association for Psychedelic Studies. Equities.com: Dr. Sue Sisley: Cannabis Medical Research is Being Kept in the Dark as Thousands Die Each Year. Available at: https://maps.org/ayahuasca-treatment/261-ayahuasca-assisted-treatment-for-addiction-british-columbia-canada/5959-equities-com-dr-sue-sisley-cannabis-medical-research-is-being-kept-in-the-dark-as-thousands-die-each-year Accessed: March 2019.
• National Academies of Sciences, Engineering, and Medicine; Health and Medicine Division; Board on Population Health and Public Health Practice; Committee on the Health Effects of Marijuana: An Evidence Review and Research Agenda. The Health Effects of Cannabis and Cannabinoids: The Current State of Evidence and Recommendations for Research. Washington (DC): National Academies Press (US); 2017 Jan 12. 15, Challenges and Barriers in Conducting Cannabis Research. Available from: https://www.ncbi.nlm.nih.gov/books/NBK425757/
• National Institute on Drug Abuse. NIDA’s Role in Providing Marijuana for Research. Available at: https://www.drugabuse.gov/drugs-abuse/marijuana/nidas-role-in-providing-marijuana-research Accessed: March 2019.
• Stith SS & Vigil JM. Federal barriers to cannabis research. Science 2016;352(6290):1182. doi: 10.1126/science.aaf7450. http://science.sciencemag.org/content/352/6290/1182.1.long
• The Guardian (2018). UK doctors will be able to prescribe cannabis medicine next month. Available at: https://www.theguardian.com/society/2018/oct/11/uk-to-allow-doctors-to-prescribe-cannabis-based-medicine-epilepsy-sajid-javid Accessed: March 2019.
• Wired (2019). Medical cannabis is now legal in the UK, but nobody can get a prescription. Available at: https://www.wired.co.uk/article/medical-cannabis-nhs-prescription Accessed: March 2019.

Pittcon Tracks

Chapter 2 – What’s in this Cannabis Product?

As laws around cannabis use relax, there is a growing need for analytical tools that can ascertain the quality, origins and purity of cannabis products in both the recreational and medical settings. Cannabis products have the potential to become both deliberately and inadvertently contaminated, and can contain harmful substances such as heavy metals, pesticides, residual solvents and pathogens. Verifying the authenticity and potency of cannabis products is particularly important when they are being used for therapeutic purposes to ensure delivery of the active substance and correct dosing.

Cannabis authenticity

NMR is one technique that has a lot to offer in the field of cannabis analysis, including use in verifying the authenticity and origins of a cannabis product.

In a recent paper, Harrington and Wang used NMR technology from Bruker, an exhibitor at this year’s Pittcon, to analyze botanical samples, using tea extracts, liquor samples, hop and cannabis extracts. They showed that peaks for the components of each sample were readily identified by comparing the spectra to those for six classifier compounds. Techniques such as this have applications not just in the field of cannabis research and regulation but also in the nutraceutical industry, where natural products are sold alongside health claims, but often without verification of their authenticity or efficacy.

Pattern recognition can also be used to make NMR analysis of cannabis samples more rapid. Creating an NMR “fingerprint” of cannabis’ chemical profile means that traits such as growing location, strain, hybrid or quality can be determined by the presence of key peaks.

Heavy metal contamination

As plants grow they absorb metals from the water and surrounding soil. In some cases these are beneficial for human consumption, such as the concentration of iron in leafy green vegetables. However, many metals are harmful to human consumption. Examples include cadmium, lead, mercury and arsenic (the “big four”).

At Pittcon 2019, Andrew Fornadel from Shimadzu Scientific Instruments, will provide detail on analytical workflows for metal testing in cannabis products using inductively coupled plasma mass spectrometry (ICP-MS).

A team from Shimadzu have recently shown how the company’s ICPMS-2030 device is able to determine heavy metal concentrations by spiking cannabis flower samples with 5 parts per billion cadmium, lead, mercury and arsenic. The results demonstrated that the technology is able to detect heavy metals with sensitivity and accuracy that meets or exceeds current regulatory standards. The researchers note that this provides flexibility that should regulations on contaminant concentrations become more stringent with time.

Also exhibiting at Pittcon 2019, will be Advion, who also offer an ICP-MS device suitable for cannabis testing laboratories.

Shimadzu have brought to market a dedicated cannabis analyzer, which is intended to make cannabis analysis more accessible to non-specialists. It is able to determine the potency of a cannabis sample through quantitation of the major cannabinoids, and can be used on samples of a variety of origins, such as whole-plant marijuana, edibles and waxes.

Pesticides

Jack Henion from Advion, will also be giving a symposium presentation on the use of single quadrupole mass spectrometry to determine the potency and detect pesticides in cannabis products.

He will present results of performing this MS method with three different API inlet systems, including liquid-chromatography/mass spectrometry for a full-scan exploratory analysis of a sample that contains unknown constituents, as well as targeted quantitation of cannabinoids and pesticides. He will also cover rapid screening analyses, including a method harnessing atmospheric pressure chemical ionization and another using open port source ionization as a simple and fast method to detect cannabinoids.

Also exhibiting at Pittcon 2019 will be ThermoFisher, Waters and Shimadzu, who also offer single quadrupole mass spectrometer products.

Pittcon 2019 will also feature a talk from Kevin A Schug of the University of Texas. His lab carries out much of their research using hops as a proxy for cannabis due to legal restrictions within the state of Texas. He will outline methods that provide a “toolbox” for a more concise characterization of natural cannabis products, primarily focused on terpenes and cannabinoids. These approaches include headspace gas chromatography, dual vacuum ultraviolet-triple quadrupole detection and tandem mass spectrometry employing various inlet systems for cannabinoid identification. Such methodologies also have potential applications in the detection of cannabis metabolites in biological fluids which could have utility for law enforcement.

At Pittcon 2019 you have the opportunity to meet representatives from Waters who have a range of products for tandem quadrupole mass spectrometry.

References

• Analytical Cannabis (2017). NMR Spectroscopy: producing a chemical fingerprint of cannabis. https://www.analyticalcannabis.com/articles/nmr-spectroscopy-producing-a-chemical-fingerprint-of-cannabis-292728 Accessed: March 2019.
• Davis D, Long K, Masone J et al. (2015) Analysis of “The Big Four” Heavy Metals in Cannabisby USN-ICP-OES. Available at: https://www.shimadzu.co.uk/sites/shimadzu.seg/files/analysis_of_heavy_metals_in_cannabis_by_usn_icp_oes.pdf Accessed: March 2019.
• Harrington P & Wang X. Spectral Representation of Proton NMR Spectroscopy for the Pattern Recognition of Complex Materials. J Anal Test 2017: DOI: 10.1007/s41664-017-0003-y.
• News Medical (2018) Cannabis quality and contamination testing. Available at: https://www.news-medical.net/news/20180205/Cannabis-Quality-and-Contamination-Testing.aspx Accessed: March 2019.
• Pruszkowski & Bosnak C (2017). The Benefits of ICP-MS for the Determination of Toxic and Nutritional Elements in the Cannabis Family of Flowering Plants. Available at: http://www.chromatographyonline.com/benefits-icp-ms-determination-toxic-and-nutritional-elements-cannabis-family-flowering-plants Accessed: March 2019.
• Shimadzu (2017). Analysis of Heavy Metal Contaminants in Cannabis Flower using the Shimadzu ICPMS-2030. Available at: https://www.shimadzu.eu/sites/shimadzu.seg/files/006_cannabisicpms.pdf Accessed: March 2019.
• Shimadzu. Cannabis analyzer for potency. Available at: https://www.ssi.shimadzu.com/products/liquid-chromatography/cannabis-analyzer.html Accessed: March 2019.
• The Resonance. Using proton NMR spectroscopy to authenticate botanical extracts. Available at: https://www.theresonance.com/using-nmr-to-authenticate-botanical-extracts/ Accessed: March 2019.
• Waters. Tandem quadrupole (triple quadrupole) mass spectrometry. Available at: http://www.waters.com/waters/en_GB/Tandem-Quadrupole-%28-Triple-Quadrupole-%29-Mass-Spectrometry/nav.htm?cid=134656172&xcid=ext4979&gclid=EAIaIQobChMI5d-3p6fQ4AIVqrDtCh3BEwT2EAAYASAAEgKN0PD_BwE&locale=en_GB Accessed: March 2019.

Chapter 3 – Improving Cannabis Safety

“Without clear, consistent, enforced quality control standards, the current legal medical cannabis industry offers little consumer advantage over the black market… Thus, there is a dire need for the development and implementation of standards for the cultivation, processing, labeling, and quality assurance testing of cannabis products.” – Dr Ryan Vandrey, Johns Hopkins University.

There are inherent challenges to bringing a botanical substance to consumers and supplying it in a safe and consistent form. This is no less true for cannabis, particularly with regards to medical cannabis. When cannabis is to be provided in a medical setting then it must reach the same standards as other pharmaceutical therapies.

Delivering a safe and consistent product requires good practice at every stage, from “seed-to-sale”. The characteristics of cannabis vary according to the particular strain and the conditions it grows in. Best practice would ensure hygienic procedures to minimize the microbiological load of the cannabis crop, while optimal drying, ventilation and storage prevent mold growth and product degradation.

It is also vitally important to verify the potency of cannabis and in particular the ratio of THC to CBD. For medical marijuana, a high-CBD, low-THC ratio is desired to maximize therapeutic effects while minimizing unwanted psychoactive side effects. A 2015 report by the US Food and Drug Administration (FDA) found that there were products on the market which claimed to contain CBD that, on testing, had none.

As with other pharmaceutical products there are also questions about how the delivery of the drug alters its properties and dosing. For example, whether it is combined with excipients, and whether it is ingested or inhaled.

There is also an onus on those in the cannabis production chain to implement their own in-house testing to more rapidly identify and deal with problems. Currently most testing is carried out on the end product at independent testing labs. However, with advances in technology, cannabis analysis no longer requires vast space for equipment or extensive user experience, making it increasingly practical to implement for cultivators, processors and retailers.

In the United States, although recreational or medical use has been authorized by several states, cannabis is still illegal at the federal level. Consequently, the FDA does not recognise cannabis as a regulated product and does not provide any standards for its cultivation and supply, meaning it is up to states that have opted to make legal provision for cannabis to create and enforce their own standards. As a result, there is no standardization of testing between states in the USA.

At Pittcon 2019, Lori Dodson of the Maryland Medical Cannabis Commission (MMCC) will describe how her state have developed regulations and standards for their licenced production and dispensing program for medical cannabis, under these circumstances.

The bill permitting medical marijuana within Maryland was signed into law in 2014 but the program did not get up and running until 2017. It was reported that during the first year, there were 2 million cannabis transactions by dispensaries. MMCC is responsible for licencing medical marijuana growers, processors and dispensaries, approving independent testing labs and registering patients with eligible medical conditions.

Dodson, the organization’s deputy director and director of lab compliance will discuss the regulations for independent testing labs established by MMCC to ensure the safety and potency of medical cannabis. She will cover the tests required for cannabis products and the establishing of acceptable tolerance limits for medical cannabis products such as for pesticides, residual solvents and biological impurities.

Also speaking at Pittcon 2019 is Jacklyn R Green, chief executive officer of Agate Biosciences. She will describe the pitfalls of trying to bring safe and consistent cannabis to sale throughout the many intermediate stages of “seed-to-sale”. Her company have developed the Cannabis Systems Engineering Toolkit which harnesses techniques such as system architecture, predictive modeling, and visual analytics to bring a systems engineering approach into cannabis product development. Green believes that this kind of integrative approach is needed by the cannabis industry to bring it into the mainstream while addressing the rigors of stringent regulation.

References

• Analytical Cannabis (2019) The Importance of Quality Control in the Cannabis Industry. Available at: https://www.analyticalcannabis.com/articles/the-importance-of-quality-control-in-the-cannabis-industry-311441 Accessed: March 2019.
• Marijuana Business Daily (2019) Chart: Maryland’s first-year medical marijuana sales approach $100 million. Available at: https://mjbizdaily.com/chart-marylands-first-year-medical-marijuana-sales-approach-100-million/ Accessed: March 2019.
• MMCC (2017) The Natalie M. Laprade Maryland Medical Cannabis Commission (MMCC) Technical Authority For Medical Cannabis Testing. Available at: https://mmcc.maryland.gov/Documents/Final%20Draft%20MMCC%20Technical%20Authority.pdf Accessed: March 2019.
• Norml (2014) Maryland: Marijuana Law Reform Measures Signed Into Law. Available at: https://norml.org/news/2014/04/17/maryland-marijuana-law-reform-measures-signed-into-law Accessed: March 2019.
• Thomas BF, ElSohly, MA. Chapter 5 – Quality Control and Stability Assessment, Editor(s): Brian F. Thomas, Mahmoud A. ElSohly, The Analytical Chemistry of Cannabis, Elsevier, 2016, Pages 83-99, ISBN 9780128046463. https://doi.org/10.1016/B978-0-12-804646-3.00005-9.

Chapter 4 – Implications for the Future

The lack of standardization in cannabis production and safety is a major obstacle to the expansion of the field of cannabis research and bringing approved products to market. Addressing this issue will be an important next step for the industry.

At Pittcon 2019, Katherine K Stenerson from Merck (formerly Millipore Sigma) will present a talk on how to improve cannabis testing workflows. She will discuss the workflows for several types of test that are usually performed on cannabis, including potency, for pesticide residue and terpene profiling, and will also explore how the quality of testing can be improved.

Stenerson will discuss the use of solid phase microextraction (SPME) sample preparation for terpene analysis. This method is now in widespread use for many applications, such as in analysing flavors and fragrances and has been used extensively in the extraction of volatile and semi-volatile organic pollutants from water.

Stenerson has previously used the technique to show how the cannabinoids can be quantified from water samples with this method when combined with a derivitization process prior to analysis by GC/MS. Such a method could be a more reliable way to measure cannabis usage from sewage, rather than self-reported surveys.

The SPME method involves use of a silica fiber that is coated with a liquid, solid or combination of both. The fibre directly extracts analytes from the sample without the need for a solvent, before they can be inserted directly into the chromatograph for analysis. The approach has a number of advantages, including simplicity and a relatively short processing time.

Cannabis has been most commonly associated with recreational use in modern culture but more and more people are turning to it for medicinal purposes, in some cases, risking prosecution in order to obtain the drug. Pittcon 2019 will hear from one mother who first turned to cannabis in desperation when her baby daughter was diagnosed with a brain tumor and has gone on to become involved in pre-clinical research into the role of cannabis in cancer treatment through her non-profit organization Saving Sophie.

To date, the role of cannabis and cannabinoids in cancer care has been primarily restricted to palliation. However, there is an increasing body of evidence from pre-clinical and clinical trials that suggest it could have therapeutic actions including anti-tumor activity. For example, cannabinoids have been demonstrated to induce the process of autophagy (cell digestion) in glioma, melanoma, hepatic, and pancreatic cancer. However, the American Cancer Society notes that studies have not shown that cannabinoids can control or cure cancer.

A potential benefit of cannabis and cannabinoids in cancer and other medical conditions is on tolerability and consequently patient adherence. This is particularly so in cancer where many conventional therapies are accompanied by severe side effects, and some research has indicated that smoking cannabis can help reduce the severity of chemotherapy-induced nausea and vomiting.

Other areas of interest in clinical research include the treatment of chronic pain and epilepsy. Some studies have shown that smoked or inhaled cannabis can improve symptoms of chronic or neuropathic pain.

Some cannabinoid therapies have been approved by the US FDA for use in HIV/AIDS-induced anorexia and chemotherapy-induced nausea and vomiting in cancer patients, while certain forms are an extracted form of CBD that is available for prescription to children with two types of treatment-resistant epilepsy known as Dravet and Lennox-Gastaut syndrome.

References

• American Cancer Society. Marijuana and Cancer. Available at: https://www.cancer.org/treatment/treatments-and-side-effects/complementary-and-alternative-medicine/marijuana-and-cancer.html Accessed: March 2019.
• Food & Drug Administration (2018). FDA approves first drug comprised of an active ingredient derived from marijuana to treat rare, severe forms of epilepsy. Available at: https://www.fda.gov/newsevents/newsroom/pressannouncements/ucm611046.htm Accessed March 2019.
• Kataoka, H, Lord HL, Pawliszyn J, SOLID-PHASE MICROEXTRACTION | Biomedical Applications, Editor(s): Ian D. Wilson, Encyclopedia of Separation Science, Academic Press, 2000, Pages 4153-4169, ISBN 9780122267703.
• Merck. Solid Phase Microextraction (SPME). Available at: https://www.sigmaaldrich.com/analytical-chromatography/sample-preparation/spme.html Accessed: March 2019.
• Ouyang G, 6 – Calibration, Editor: Janusz Pawliszyn, Handbook of Solid Phase Microextraction, Elsevier, 2012, Pages 167-199, ISBN 9780124160170.

Conclusion

The cannabis industry is going through a period of rapid change that involves and interests parties from many different fields and walks of life. This could not be better reflected by the cannabis analysis program at Pittcon 2019, where opening speaker Joshua Crossney will appeal for greater collaboration and cross-disciplinary activities to improve medical cannabis research and access. The program brings together speakers from a diverse number of fields, including medical research, chemistry, systems engineering and patient advocacy.

Analytical testing has a major role to play as cannabis products become more widely legalized and the Pittcon 2019 expo will be attended by all the major industry players in this area. Confirmed exhibitors include Shimadzu, Bruker, Phenomenex. Advion and Merck (previously known as Millipore Sigma). These companies offer a full spectrum of solutions for the analytical cannabis arena from sample preparation, NMR spectroscopy, ICP-MS to single quadrupole and tandem mass spectrometry.

Pittcon 2019 is taking place at the Pennsylvania Conference Center in Philadelphia, March 17-21. Register now to be a part of it.