Poster Session Abstracts and Presenters

Presenter(s): Mrinal Kanti Chatterjee², Shonali Paul², Arun Apte¹, Aditi Gade²

¹CloudLIMS, DE, USA

²CloudLIMS Software Solutions, Indore, MP, India

Introduction

Cannabis has the power to treat a myriad of medical ailments. The entire cannabis industry, from seed-to-sale, is exponentially expanding due to the legalization of cannabis in the majority of the United States. This has accelerated the growth of cannabis testing laboratories. The laboratories perform a diverse array of tests such as potency tests, cannabinoid and terpenoid profiling, microbiological screening, quantified pesticide tests, residual solvent analysis, etc. Cannabis testing laboratories encounter multitudinous challenges, which includes managing a large number of cannabis samples, multi-analyte tests, and their associated data, generating a certificate of analysis (COA), reducing turnaround time, meeting regulatory guidelines such as ISO 17025, GLP, 21 CFR Part 11, and ensuring strict quality control. Paper-based data management practices are antiquated and cannot keep pace with the increasing influx of samples, diverse testing requirements, in addition to the evolving regulatory requirements. A Laboratory Information Management System (LIMS) can play a vital role in addressing the data management challenges, maximizing automation, and in complying with the regulatory requirements.

Description: A LIMS serves as a central data repository for cannabis testing laboratories. It enhances automation by generating barcodes/RFID labels for seamless sample tracking, ensures data integrity by integrating with analytical instruments, and helps laboratories generate customized test reports and certificates of analysis. Besides, it can help laboratories meet the documentary requirements of ISO 17025, maintain an automated audit trail, and assure complete traceability by maintaining Chain of Custody (COC) of samples and tests.

Conclusion

Amongst the various LIMS delivery models, “in-the-cloud” LIMS is the most cost-effective model. A cloud-based LIMS is a scalable data management solution and can automate cannabis testing workflows, right from sample accessioning & tracking to report generation. Additionally, a cloud-based LIMS is flexible, secure, and facilitates real-time sharing of analytical data.

Presenter(s): Mrinal Kanti Chatterjee², Shonali Paul², Arun Apte¹, Aditi Gade²

¹CloudLIMS, DE, USA

²CloudLIMS Software Solutions, Indore, MP, India

Introduction

The legalization of cannabis for medicinal and recreational use, sale, and distribution in the United States is driving the growth of cannabis testing laboratories. The most prevalent botanical marijuana species are Cannabis sativa and Cannabis indica, the major source of cannabinoids. Cannabis testing laboratories test cannabis and its infused products to determine the concentration of cannabinoids, moisture, heavy metals, residual solvents, pesticides, and to screen pathogens such as Escherichia coli, for safeguarding public health. The testing laboratories face numerous challenges on a day-to-day basis such as managing all data associated with samples and tests, enhancing efficiency, meeting regulatory compliance such as ISO 17025, GLP, 21 CFR Part 11 (electronic signatures), reducing operational expenses, and meeting quality standards. A Laboratory Information Management System (LIMS) effectively addresses these challenges, as opposed to spreadsheets and other paper-based data management methods. Owing to the benefits offered by cloud technology, a cloud-based LIMS is more cost-efficient and easier to run when compared to a traditional on-premise LIMS.

Description: A LIMS serves as a controlled database to manage comprehensive sample and test data. It helps automate cannabis testing workflows including test result reporting and generation of Certificates of Analysis (CoA). A LIMS records the Chain of Custody (CoC) for samples and tests while maintaining an automated audit trail. It also facilitates auto-scheduling of instrument calibration and inventory expiration/depletion alerts to comply with regulatory requirements. Furthermore, it helps maintain Standard Operating Procedures (SOPs) for laboratory personnel and meets state accreditations.

Conclusion

A cloud-based LIMS is a technologically-advanced and a cost-effective alternative to a traditional on-premise LIMS. A cloud-based LIMS leverages the advantages of cloud technology, enabling testing laboratories to forgo IT requirements, reduce the total cost of ownership, and encourage collaboration among cultivators, manufacturers, and retailers of cannabis products.

Presenter(s): Brandon Canfield, PhD, Associate Professor of Chemistry, Lesley Putman, PhD, Professor of Chemistry, Northern Michigan University, MI, USA

A new undergraduate degree program has been developed at Northern Michigan University to specifically focus on the chemistry associated with the cannabis industry. The first program of its kind, this degree requires extensive coursework in the fields of chemistry and biology, with advanced instruction and instrumentation applied toward direct analysis of plant secondary metabolite compounds in raw plant material and in manufactured products, as well as additional agricultural chemical analyses, such as inorganic nutrients and pesticide residue. The full four-year curriculum will be presented, with explanations for the decisions made throughout the program design process, including the decision to include an entrepreneurial track option with additional coursework in accounting, finance, and marketing. Finally, the experience, skills, and knowledge which can be expected from our graduates will be summarized.

Presenter(s): Chris Denicola¹, Joe Barendt².

¹Prism Scientific, CO, USA, ²Chiral Technologies, Inc., PA, USA

Cannabinoid isolation and purification is getting a lot of attention from the medical and recreational marijuana and hemp industries.  Flash liquid chromatography provides a cheap and cost effective entry into purification but the solvent usage and evaporation can be more than most companies can handle.  Supercritical Fluid Chromatography may not be the most common purification method but, when combined with immobilized chiral stationary phases, can be very powerful.  The goal of this work is to show that for isolating specific cannabinoids, utilizing immobilized CSP greatly increases throughput. Through the use of standards and real world samples, we will show how to isolate specific cannabinoids with an eye on throughput, loading, solvent usage and solvent recycling.  We will also show that the use of immobilized CSPs are preferred over traditional CSPs due to their ruggedness and longevity.  Marijuana and hemp extracts contain compounds that can interact with traditional CSPs and cause loss of retention, resolution, selectivity, and tailing.

Presenter(s): Johnson S¹, Speck M², Hilyard A¹, Afia I¹, Orser C¹

¹Digipath Labs, NV, USA; ²Chaminade University, HI, USA

The lack of horticultural or agronomic naming conventions or mandatory cultivar registration in the cannabis industry has created a confusing collection of strain names, making authenticity questionable for the cannabis consumer. Various chemometric-based efforts have been underway to make sense of cannabis strain names through the use of data analytics.

Terpenoids, in addition to their pharmacological activity, can be used as distinctive profiles that are statistically useful for distinguishing clusters of similar strains, or cultivars. We analyzed the chemical profiles of 2,237 individual cannabis flower samples, representing 204 individual cultivars across 27 cultivators in a tightly regulated Nevada cannabis testing market. The flower samples were analyzed across 11 cannabinoids and 19 terpenoids. Principal component analysis (PCA) of the combined dataset resulted in three distinct clusters that were distinguishable by terpene profiles alone. Further dissection of individual cultivars by cultivators within clusters revealed striking fidelity of terpene profiles and also revealed a few outliers from the dominant Type I CBDA/THCA ratio in Nevada. We propose that three terpene cluster assignments account for the diversity of drug-type cannabis strains currently being grown in Nevada.

Presenter(s): Frank A. Kero, Ph.D., Wilhad Reuter, Jason Weisenseel

PerkinElmer, CA, USA

Current trends in the analysis of the cannabinoid content in commercially available food products points towards liquid chromatography to ensure label claim accuracy in product content descriptions. This analysis can be challenging since the fortification of cannabinoid compounds has been applied to a diverse spectrum of matrices including high sugar, high fat materials. To help accommodate workflow for these products, a UHPLC has been evaluated with an autosampler modification that allows the sample to be pulled into the syringe cannula from the side versus drawing sample from the bottom of the vial. It is suspected that dirty samples that may precipitate would clog a needle drawing from the bottom. This study evaluates the system performance with several types of food matrix variables.

Presenter(s): Frank A. Kero, Ph.D.¹, Luke Ward², Ben Armstrong², Stephanie Marin³, Jacob Jalali¹, Jason Weisenseel¹, Erasmus Cudjoe¹

¹PerkinElmer, CA, USA ²Juniper Analytics, OR, USA ³Biotage, NC, USA

Recent changes in regulatory guidelines for the quality control of medicinal cannabis and cannabis for recreational use has highlighted some interesting analytical challenges in the analysis of pesticides in flower varietals. Of particular interest to this fundamental investigation is the identification of endogenous matrix components that may lead to non-specific analyte ionization suppression when analyzed by UPLC-ESI-MS/MS. This presentation will detail a 3 experiment approach to develop a better understanding of chemical noise variability from sample-to-sample. A post-column infusion experiment was performed to determine where the suppression events occur in the chromatographic run. Next, the sample was injected in the Q1 scanning operational mode to determine what masses are present during these retention time windows and a subsequent literature search was performed to correlate chemical noise identification. Since it has been well documented that high concentration of cannabinoids are present in these materials, an MRM experiment was performed to determine if these compounds correlate to the retention times of the suppression events observed in the chromatographic run. Sample preparation variables will be executed using this template to determine the significance of improvement when comparing experiment complexity and cost per sample. The UPLC for this investigation was a PerkinElmer Altus A30 UPLC. The triple quadrupole mass spectrometer was a QSight 210 fitted with an HSID source to allow for the injection of dirty samples without the need to break vacuum and clean between experiments. It is anticipated that this poster will help support method development efforts towards improved robustness and quantitative accuracy.

Presenter(s): Shawn Helmueller, Jason Hill

Waters Corporation, Milford, MA, USA

Online SFE-SFC instrumentation is used to generate real-time analytical information during batch extractions. Strategies for optimizing extraction cycles, vessel switching, and scaling from batch to bulk processes are discussed.

On-site analytics has been the standard for decades in the food and beverage, nutraceutical, and pharmaceutical industries, but it is not yet standard across the cannabis industry. As a result, processors performing extractions and formulating high value-added products may potentially suffer from missing, incomplete, or delayed analytical results; and this delay in analytical information can lead to quality issues, and costly inefficiencies in the development and execution of processing workflows. This talk will highlight a two dimensional SFE-SFC system that moves extract spectral and chromatographic analysis online, to the point where the extract is first generated in the extraction cycle. This real-time extraction analytical information has the potential to quickly identify and correct inefficiencies in the extraction and processing workflow, while streamlining the tedious optimization/re-optimization processes involved in developing targeted extraction outcomes.

Presenter(s): Jacqueline Harding

Cannabistry Labs, CO, USA

Cannabis extracts are evaluated primarily by the potency, color, yield, and the capacity to mimic the natural flower profile. The outcome of the final extract product is highly dependent upon the extraction method and subsequent post processing techniques. The use of high temperatures during distillation results in the destruction of the terpene profiles and the use of hydrocarbons as extraction solvents results in the coextraction of undesirable plant components and a limited capacity for extraction optimization. The research presented here will examine the optimization of supercritical fluid CO2 extractions and subsequent post processing methodologies to produce high grade cannabinoid extracts. The extraction processes we examine will emphasis the capture of terpenes that mimic the natural profile of the flower and cannabinoid extractions exceeding 95% extraction of cannabinoids from the plant matter. Postprocessing methods utilized here emphasize the removal of coextraction products including chlorophyll. The use of our methodology is shown to result in the complete removal of coextracted chlorophyll without requiring high temperature distillation and minimal loss of cannabinoids. The resulting extract is a translucent amber color with 3rd party laboratory potency testing as high as 94% Max THC. The methodologies described here facilitate the preparation of high grade oil extracts that can be directly utilized in formulated products including vape cartridges without the need for non-natural diluents. As a result, formulated products can be implemented without the need of flavor masking agents and can be tuned to mimic the natural flavor and efficacy profiles of cannabis flower.

Presenter(s): Joan Stevens¹, Lilly Asanuma², Rick Jordan²

¹Agilent Technologies, DE, USA ²Pacific Agricultural Laboratory, OR, USA

Cannabis flower is a very complex matrix which makes the analysis of pesticides quite challenging. The complexity of the cannabis matrix is shown in the unbalance between cannabinoid content and pesticide level. The cannabinoids are produced at 10-20% levels or greater which correlates to 100,000-200,000 ppm whereas the pesticides even at 100 ppb is only 1.0 x 10-5 %. Other compounds like terpenes and lipids produced at 0.001-0.5% (10-5000 ppm) can also cause issues with mass spectrometers as they move through the system depositing on the flow path of the instrument. Many sample preparation approaches have been used from simple dilution, QuEChERS methodology through to solid phase extraction. Removing enough matrix to analyze the pesticides and reducing the deleterious effects on the system is crucial. This study took a practical look at the effect sorbents and sorbent mixes had on matrix removal and pesticide recoveries in a dispersive SPE (dSPE) format. One gram of homogenized cannabis material was extracted with acetonitrile. The extract was subjected to a systematic approach where sorbents were incrementally increased and various sorbent mixtures were formulated. This study is a guide to better understand the effect various sorbents have on pesticide recovery in cannabis flower. Criteria for acceptable pesticide recoveries are 70-120% and S/N ≥ 10. Pesticide recovery for the various dSPE formulations were evaluated by triple quadrupole LC/MS using a superficially porous column and on-line dilution.

Presenter(s): Joan M. Stevens and Sue D ’Antonio

Agilent Technologies, DE, USA

A cannabis edible, also called cannabis-infused food, is a food product that contains cannabinoids, especially THC and CBD. Cannabis edibles are consumed for both medical and recreational purposes. Because cannabinoids are soluble in lipids and alcohols, cannabis must be cooked with one of these two substances in order to infuse the cannabinoids into the food. The oil-solubility of cannabis extracts has been known since ancient times. Since the infused cannabinoids are a concentrate it is important to monitor chemical residues in the concentrate, specifically pesticides. However, it can be difficult to analyze for pesticide residues in high lipid content matrix. Lipids can cause both analysis and instrument issues over time. QuEChERS and organic extractions are commonly used sample preparation methods for the analysis of pesticides from food products however the dispersive SPE containing C18 which is used for fatty matrix is insufficient at selectively removing lipids. A novel sorbent Enhanced Matrix Removal-Lipid (EMR-Lipid) specific for the removal of lipids has shown to be very effective at retaining lipids without compromising recovery of the pesticides. The application will show the matrix removal capabilities of newly formulated EMR-Lipid specific for high lipid content cannabis-infused food products and the analysis of pesticides.

Presenter(s): Justin Steimling, Ashlee Reese,

Restek Corporation, PA, USA

More than 100 cannabinoids have been isolated from cannabis in addition to the five most commonly tested:  THC, THCA, CBD, CBDA, and CBN.   While methods have been published that show the separation of these major cannabinoids, many do not take into account the possibility of interference from other cannabinoids that may be present.  This is most problematic in concentrates where minor cannabinoids can be enriched to detectable levels that were not observed in the flower.  Additionally, some terpenes have been shown to absorb UV light at 228 nm, the wavelength cannabinoids are typically detected, which can result in an additional source of interference.  In this study, the LC-UV separation of 15 cannabinoids of interest was performed while monitoring for the potential impact from minor cannabinoids and terpenes on reported potency values.  The method is applied to commercially available CBD oils that have recently become suspect due to inaccurate label claims.

Ken Tseng¹, Toshi Ono¹, Tsunehisa Hirose²

¹Nacalai USA, Inc., CA, USA ²Nacalai Tesque, Inc., Kyoto, Japan

A mixture of 11 cannabinoids was separated under 14 minutes using isocratic MS-compatible mobile phases on a core-shell PBr HPLC column with UV detection. CBDV, THCV, CBDA, CBD, CBG, CBGA, CBN, Δ9-THC, Δ8-THC, CBC, and THC-A standards were mixed in a solvent of 1:1 water:methanol to a concentration of 9.1 μg/mL. 5μL injection volume was used to obtain the chromatogram. The isobaric Δ9-THC and Δ8-THC was baseline separated. The peak shapes are symmetrical for accurate quantification.

Another reversed-phase column, Cholester, is used to differentiate Δ9-THC from Δ8-THC at shorter time than with PBr.

Presenter(s): Mallory Speakman, Josh Wurzer, Travis Ruthenburg

SC Labs, CA, USA

Chemical contaminants have been commonly used during the production of cannabis, which include insecticides, fungicides, and plant growth regulators.  The state of California has set guidelines to limit these chemical residues in an effort to regulate clean and safe products for the adult-use and medicinal-use market.  Pesticides pose different health risks for inhaled cannabis products, so strict guidelines were set.  A comprehensive list with low action limits has given the cannabis market a challenge that they seem to have risen to.  Long-term ongoing analysis of pesticide data has been collected for annual cannabis competitions.  Pesticides are analyzed by High Performance Liquid Chromatography (HPLC) using a triple quadropole mass spectrometer with electrospray ionization.  Year over year data has shown largely a decrease in the number of pesticide detections.  SC Labs will be releasing new pesticide data collected from 2017 for cannabis flower and concentrate samples, emphasizing the decreasing trend over the past couple years.  As we move into the upcoming year with a regulated market, we hope to continue to see this decreasing trend thrive for safe cannabis.

Presenter(s): Melissa Wilcox¹, Giulia Mazzoccanti², Omar H. Ismail², Alessia Ciogli², Claudio Villani², Francesco Gasparrini²

¹Regis Technologies, IL, USA ²Dipartimento di Chimica e Tecnologie del Farmaco – Sapienza Università di Roma, Roma, Italy

The interest in the medical use of Cannabis Sativa L. is steadily increasing because of its therapeutic efficacy towards a wide variety of ailments, and its unique chemistry, characterized by the presence of cannabinoids that are concentrated in the female inflorescence. The fiber type of Cannabis Sativa L. is cultivated in Europe for textile production or for food (seeds, flour, and oil), and has a low concentration of psychoactive (-)-Δ9-trans-tetrahydrocannabinol ((-)-Δ9-THC) that is typically less than 0.2%. The main cannabinoid in the fiber type of Cannabis Sativa L. is (-)-cannabidiol ((-)-CBD) but there is also (-)-cannabidivarin ((-)-CBDV), cannabigerol (CBG), cannabinol (CBN) and the racemic cannabichromene (rac-CBC), each having various therapeutic actions. The analysis of the original composition of plant material is necessary for phenotype determination and quality control of medicinal cannabis used in therapeutic treatments.

The presence of natural racemic compounds (rac-CBC) in plant extract was investigated using Chiral Stationary Phases (CSPs) in enantioselective “e” Ultra High Performance Supercritical Fluid Chromatography (eUHPSFC). All of the analyses were performed using a UHPSFC-compatible chiral column developed in the Sapienza University laboratory in collaboration with Regis Technologies.  Kinetic evaluation using van Deemter plots showed excellent kinetic performance for the 100×4.6 mm column packed with 1.8 µm on a modified low-dispersion Waters UPC2 system. Using a 90:10 CO2/MeOH mixture as mobile phase and TSO (trans-Stilbene oxide) as a probe, efficiencies up to 290,000 N/m were measured on the first enantiomer at flow rate of 4.0 mL/min, and more than 278,000 N/m on the second enantiomer (flow rate = 3.7 mL/min) were observed .  The Chiral Stationary Phase (CSP) allowed resolution of the racemic compound cannabichromene (rac-CBC) in plant extract and the synthetic racemic (+/-)-Δ9-trans-tetrahydrocannabinol. Good separation, in terms of chemio- and enantio- selectivity, was obtained with high resolution for all cannabinoids, and their acid forms, under isocratic conditions.

Presenter(s): Patricia L. Atkins, Sean Curran

SPEX CertiPrep, NJ, USA

The cannabis industry has taken the scientific world by storm, flooding the market with new products. Recently, concerns have arisen about safety of this, unregulated market, resulting in many new labs testing for cannabinoid potency, pesticides, bacteria/mold and other potential contaminants.  Sadly, a potentially significant group of contaminants has been largely ignored:  toxic metals.  Recreational cannabis and hemp are part of the C.sativa species, with different cultivars resulting in unique cannabinoid profiles. Federally legal hemp products (hempseed oil, hemp extracts, CBD oil & extracts), those not containing the psychoactive THC, are widely available on the market today. Such products are also used as a base oil for the addition of cannabis & cannabinoid extracts (including medical & recreational cannabis products). However, due to a ban on it’s cultivation in the US, virtually all of the hemp used is imported from China, India, & Eastern Europe. Studies of other consumable commodities exported from these countries have reported widespread heavy metal contamination (i.e. spices, teas, grains etc.).

Cannabis plants (hemp & recreational varieties) are bio-accumulators of heavy metals.  In the production of many of the above mentioned products, a large amount of plant material is processed to extract concentrates and oils, thereby increasing the risk of heavy metal contamination.  The scope of this study was twofold; firstly, to analyze various legal hemp products currently on the market and, secondly, to use these as a model for methods development for testing of restricted products.  Samples were digested using microwave digestion and analyzed by ICP-OES and ICP-MS to determine the elemental composition of these products and the concentration of potentially toxic heavy metals. A large number of the tested hemp products detected heavy metal contamination.

Presenter(s): Fritz Chess¹, D.J. Gold²

¹Eden Labs, WA, USA ²Author, Cannabis Alchemy

The program will begin with a description of distillation/extraction methods and the history and technical relevance of the medieval Connelly still, D.J. Gold’s designs, and the soxhlet extractor will be explained. This will be followed by the history and evolution of steam distilling and hydro distilling and will be explored from a perspective of modern methods of steam distilling under deep vacuum to make low temperature steam. Vacuum steam distilling reduces thermal degradation of essential oils or mono terpenes so they retain their aroma and integrity. In conclusion, I will explain how these two methods have been combined in one machine so that light terpenes can first be harvested and then various types of ethanol extractions of cannabinoids containing oleoresins can then be performed.
This presentation will include slides of extractors, soxhlet extractors, and steam distillers.

Presenter(s): Travis Kennedy, Blake Grauerholz, Dr. Markus Roggen

OutCo Labs, CA, USA

Our work at OutCo on optimization of supercritical CO2 extraction has widely been reported. We now have been turning out attention on other forms of cannabis concentrate production. Rosin has been around for a few years in the cannabis industry. Initially started by utilizing hair irons, the technology moved on to hydraulic or air-actuated heated metal plates. This allows the oil producer to control a plethora of factors. With more sophisticated rosin presses coming to the market, we saw the need to investigate which factors have the largest influence on oil yields and quality. We utilized Design of Experiment protocols and developed a quality grading system to quickly study a variety of factors in rosin pressing. With this poster, we want to present our latest findings and further a discussion on process optimization.

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