Tag Archives: analytical

IR Spectrum of 2,4-Dichlorophenol in different physical states
From The Lab

Gas Chromatography/Infrared Spectroscopy: A Tool For the Analysis of Organic Compounds in Cannabis

By John F. Schneider
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IR Spectrum of 2,4-Dichlorophenol in different physical states

Editor’s Note: The author will be teaching a 1/2 day short course on this topic at PITTCON in Philadelphia in March 2019.


The combination of gas chromatography and infrared spectroscopy (GC/IR) is a powerful tool for the characterization of compounds in complex mixtures. (1-5) Gas chromatography with mass spectroscopy detection (GC/MS) is a similar technique, but GC/MS is a destructive technique that tears apart the sample molecules during the ionization process and then these fragments are used to characterize the molecule. In GC/IR the molecules are not destroyed but the IR light produced by molecular vibrations are used to characterize the molecule. IR spectrum yields information about the whole molecule which allows the characterization of specific isomers and functional groups. GC/IR is complementary to GC/MS and the combination results in a powerful tool for the analytical chemist.

A good example of the utility of GC/IR vs GC/MS is the characterization of stereo isomers. Stereo isomers are mirror images such as a left hand and a right hand. In nature, stereo isomers are very important as one isomers will be more active then its mirror image. Stereo isomers are critical to medicinal application of cannabis and also a factor in the flavor components of cannabis.

GC/MS is good at identifying basic structure, where GC/IR can identify subtle differences in structure. GC/MS could identify a hand, GC/IR could tell you if it is a left hand or right hand. GC/MS can identify a general class of compounds, GC/IR can identify the specific isomer present.

Why GC/IR?

Gas chromatography interfaced with infrared detection (GC/IR), combines the separation ability of GC and the structural information from IR spectroscopy. GC/IR gives the analyst the ability to obtain information complementary to GC/MS. GC/IR gives the analyst the power to perform functional group detection and differentiate between similar molecular isomers that is difficult with GC/MS. Isomer specificity can be very important in flavor and medical applications.

 IR Spectrum of 2,4-Dichlorophenol in different physical states

IR Spectrum of 2,4-Dichlorophenol in different physical states

Gas chromatography with mass spectrometry detection (GC/MS) is the state-of-the-art method for the identification of unknown compounds. GC/MS, however, is not infallible and many compounds are difficult to identify with 100 % certainty. The problem with GC/MS is that it is a destructive method that tears apart a molecule. In infrared spectrometry (IR), molecular identification is based upon the IR absorptions of the whole molecule. This technique allows differentiation among isomers and yields information about functional groups and the position of such groups in a molecule. GC/IR complements the information obtained by GC/MS.

Interfaces

Initial attempts to couple GC with IR were made using high capacity GC columns and stopped flow techniques. As GC columns and IR technology advanced, the GC/IR method became more applicable. The advent of fused silica capillary GC columns and the availability of Fourier transform infrared spectrometry made GC/IR available commercially in several forms. GC/IR using a flow cell to capture the IR spectrum in real time is known as the “Light Pipe”. This is the most common form of GC/IR and the easiest to use. GC/IR can also be done by capturing or “trapping” the analytes of interest eluting from a GC and then measuring the IR spectrum. This can be done by cryogenically trapping the analyte in the solid phase. A third possibility is to trap the analyte in a matrix of inert material causing “Matrix Isolation” of the analyte followed by measuring the IR spectrum.

Infrared Spectroscopy

The physical state of the sample has a large effect upon the IR spectrum produced. Molecular interactions (especially hydrogen bonding) broadens absorption peaks. Solid and liquid samples produce IR spectra with broadened peaks that loses much of the potential information obtained in the spectra. Surrounding the sample molecule with gas molecules or in an inert matrix greatly sharpens the peaks in the spectrum, revealing more of the information and producing a “cleaner” spectrum. These spectra lend themselves better to computer searches of spectral libraries similar to the computer searching done in mass spectroscopy. IR spectral computer searching requires the standard spectra in the library be of the same physical state as the sample. So, a spectrum taken in a gaseous state should be searched against a library of spectra of standards in the gaseous state.

IR of various phases:

  • Liquid Phase – Molecular interactions broaden absorption peaks.
  • Solid Phase – Molecular interactions broaden absorption peaks.
  • Gas Phase – Lack of molecular interactions sharpen absorption peaks.
  • Matrix Isolation – Lack of molecular interactions sharpen absorption peaks.

IR Chromatograms

GC/IR yields chromatograms of infrared absorbance over time. These can be total infrared absorbance which is similar to the total ion chromatogram (TIC) in GC/MS or the infrared absorbance over a narrow band or bands analogous to selected ion chromatogram. This is a very powerful ability, because it gives the user the ability to focus on selected functional groups in a mixture of compounds.

Conclusion

Gas chromatography with infrared detection is a powerful tool for the elucidation of the structure of organic compounds in a mixture. It is complementary to GC/MS and is used to identify specific isomers and congeners of organic compounds. This method is greatly needed in the Cannabis industry to monitor the compounds that determine the flavor and the medicinal value of its products.


References

  1. GC–MS and GC–IR Analyses of the Methoxy-1-n-pentyl-3-(1-naphthoyl)-Indoles: Regioisomeric Designer Cannabinoids, Amber Thaxton-Weissenfluh, Tarek S. Belal, Jack DeRuiter, Forrest Smith, Younis Abiedalla, Logan Neel, Karim M. Abdel-Hay, and C. Randall Clark, Journal of Chromatographic Science, 56: 779-788, 2018
  2. Simultaneous Orthogonal Drug Detection Using Fully Integrated Gas Chromatography with Fourier Transform Infrared Detection and Mass Spectrometric Detection , Adam Lanzarotta, Travis Falconer, Heather McCauley, Lisa Lorenz, Douglas Albright, John Crowe, and JaCinta Batson, Applied Spectroscopy Vol. 71, 5, pp. 1050-1059, 2017
  3. High Resolution Gas Chromatography/Matrix Isolation Infrared Spectrometry, Gerald T. Reedy, Deon G. Ettinger, John F. Schneider, and Sid Bourne, Analytical Chemistry, 57: 1602-1609, 1985
  4. GC/Matrix Isolation/FTIR Applications: Analysis of PCBs, John F. Schneider, Gerald T. Reedy, and Deon G. Ettinger, Journal of Chromatographic Science, 23: 49-53, 1985
  5. A Comparison of GC/IR Interfaces: The Light Pipe Vs. Matrix Isolation, John F. Schneider, Jack C. Demirgian, and Joseph C. Stickler, Journal of Chromatographic Science, 24: 330- 335, 1986
  6. Gas Chromatography/Infrared Spectroscopy, Jean ‐ Luc Le Qu é r é , Encyclopedia of Analytical Chemistry, John Wiley & Sons, 2006
oregon

Turning the Oregon Outdoor Market into a Research Opportunity

By Dr. Zacariah Hildenbrand, Dr. Kevin A. Schug
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oregon

Much has been made about the plummeting market value of cannabis grown outdoors in Oregon. This certainly isn’t a reflection of the product quality within the marketplace, but more closely attributable to the oversaturation of producers in this space. This phenomenon has similarities to that of ‘Tulip Mania’ within the Dutch Golden Age, whereby tulip bulbs were highly coveted assets one day, and almost worthless the next. During times like these, it is very easy for industry professionals to become disheartened; however, from a scientific perspective, this current era in Oregon represents a tremendous opportunity for discovery and fundamental research.

Dr. Zacariah Hildenbrand
Dr. Zacariah Hildenbrand, chief technical officer at Inform Environmental.

As we have mentioned in previous presentations and commentaries, our research group is interested in exploring the breadth of chemical constituents expressed in cannabis to discover novel molecules, to ultimately develop targeted therapies for a wide range of illnesses. Intrinsically, this research has significant societal implications, in addition to the potential financial benefits that can result from scientific discovery and the development of intellectual property. While conducting our experiments out of Arlington, Texas, where the study of cannabis is highly restricted, we have resorted to the closet genetic relative of cannabis, hops (Humulus lupulus), as a surrogate model of many of our experiments (Leghissa et al., 2018a). In doing so, we have developed a number of unique methods for the characterization of various cannabinoids and their metabolites (Leghissa et al., 2018b; Leghissa et al., 2018c). These experiments have been interesting and insightful; however, they pale in comparison to the research that could be done if we had unimpeded access to diverse strains of cannabis, as are present in Oregon. For example, gas chromatography-vacuum ultraviolet spectroscopy (GC-VUV) is a relatively new tool that has recently been proven to be an analytical powerhouse for the differentiation of various classes of terpene molecules (Qiu et al., 2017). In Arlington, TX, we have three such GC-VUV instruments at our disposal, more than any other research institution in the world, but we do not have access to appropriate samples for application of this technology. Similarly, on-line supercritical fluid extraction – supercritical fluid chromatography – mass spectrometry (SFE-SFC-MS) is another capability currently almost unique to our research group. Such an instrument exhibits extreme sensitivity, supports in situ extraction and analysis, and has a wide application range for potential determination of terpenes, cannabinoids, pesticides and other chemical compounds of interest on a single analytical platform. Efforts are needed to explore the power and use of this technology, but they are impeded based on current regulations.

Dr Kevin Schug
Dr. Kevin A. Schug, Professor and the Shimadzu Distinguished Professor of Analytical Chemistry in the Department of Chemistry and Biochemistry at The University of Texas at Arlington (UTA)

Circling back, let’s consider the opportunities that lie within the abundance of available outdoor-grown cannabis in Oregon. Cannabis is extremely responsive to environmental conditions (i.e., lighting, water quality, nutrients, exposure to pest, etc.) with respect to cannabinoid and terpene expression. As such, outdoor-grown cannabis, despite the reduced market value, is incredibly unique from indoor-grown cannabis in terms of the spectrum of light to which it is exposed. Indoor lighting technologies have come a long way; full-spectrum LED systems can closely emulate the spectral distribution of photon usage in plants, also known as the McCree curve. Nonetheless, this is emulation and nothing is ever quite like the real thing (i.e., the Sun). This is to say that indoor lighting can certainly produce highly potent cannabis, which exhibits an incredibly robust cannabinoid/terpene profile; however, one also has to imagine that such lighting technologies are still missing numerous spectral wavelengths that, in a nascent field of study, could be triggering the expression of unknown molecules with unknown physiological functions in the human body. Herein lies the opportunity. If we can tap into the inherently collaborative nature of the cannabis industry, we can start analyzing unique plants, having been grown in unique environments, using unique instruments in a facilitative setting, to ultimately discover the medicine of the future. Who is with us?


References

Leghissa A, Hildenbrand ZL, Foss FW, Schug KA. Determination of cannabinoids from a surrogate hops matrix using multiple reaction monitoring gas chromatography with triple quadrupole mass spectrometry. J Sep Sci 2018a; 41: 459-468.

Leghissa A, Hildenbrand ZL, Schug KA. Determination of the metabolites of Δ9-Tetrahydrocannabinol using multiple reaction monitoring gas chromatography – triple quadrapole – mass spectrometry. Separation Science Plus 2018b; 1: 43-47.

Leghissa A, Smuts J, Changling Q, Hildenbrand ZL, Schug KA. Detection of cannabinoids and cannabinoid metabolites using gas chromatography-vacuum ultraviolet spectroscopy. Separation Science Plus 2018c; 1: 37-42.

Qiu C, Smuts J, Schug KA. Analysis of terpenes and turpentines using gas chromatography with vacuum ultraviolet detection. J Sep Sci 2017; 40: 869-877.

dSPE cleanups

The Grass Isn’t Always Greener: Removal of Purple Pigmentation from Cannabis

By Danielle Mackowsky
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dSPE cleanups
strains
Cannabis strains used (clockwise from top left): Agent Orange, Tahoe OG, Blue Skunk, Grand Daddy and Grape Drink

Cannabis-testing laboratories have the challenge of removing a variety of unwanted matrix components from plant material prior to running extracts on their LC-MS/MS or GC-MS. The complexity of the cannabis plant presents additional analytical challenges that do not need to be accounted for in other agricultural products. Up to a third of the overall mass of cannabis seed, half of usable flower and nearly all extracts can be contributed to essential oils such as terpenes, flavonoids and actual cannabinoid content1. The biodiversity of this plant is exhibited in the over 2,000 unique strains that have been identified, each with their own pigmentation, cannabinoid profile and overall suggested medicinal use2. While novel methods have been developed for the removal of chlorophyll, few, if any, sample preparation methods have been devoted to removal of other colored pigments from cannabis.

QuEChERS
Cannabis samples following QuEChERS extraction

Sample Preparation

Cannabis samples from four strains of plant (Purple Drink, Tahoe OG, Grand Daddy and Agent Orange) were hydrated using deionized water. Following the addition of 10 mL acetonitrile, samples were homogenized using a SPEX Geno/Grinder and stainless steel grinding balls. QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) non-buffered extraction salts were then added and samples were shaken. Following centrifugation, an aliquot of the supernatant was transferred to various blends of dispersive SPE (dSPE) salts packed into centrifugation tubes. All dSPE tubes were vortexed prior to being centrifuged. Resulting supernatant was transferred to clear auto sampler vials for visual analysis. Recoveries of 48 pesticides and four mycotoxins were determined for the two dSPE blends that provided the most pigmentation removal.

Seven dSPE blends were evaluated for their ability to remove both chlorophyll and purple pigmentation from cannabis plant material:

  • 150 mg MgSO4, 50 mg PSA, 50 mg C18, 50 mg Chlorofiltr®
  • 150 mg MgSO4, 50 mg C18, 50 mg Chlorofiltr®
  • 150 mg MgSO4, 50 mg PSA
  • 150 mg MgSO4, 25 mg C18
  • 150 mg MgSO4, 50 mg PSA, 50 mg C18
  • 150 mg MgSO4, 25 mg PSA, 7.5 mg GCB
  • 150 mg MgSO4, 50 mg PSA, 50 mg C18, 50 mg GCB

Based on the coloration of the resulting extracts, blends A, F and G were determined to be the most effective in removing both chlorophyll (all cannabis strains) and purple pigments (Purple Drink and Grand Daddy). Previous research regarding the ability of large quantities of GCB to retain planar pesticides allowed for the exclusion of blend G from further analyte quantitation3. The recoveries of the 48 selected pesticides and four mycotoxins for blends A and F were determined.

dSPE cleanups
Grand Daddy following various dSPE cleanups

Summary

A blend of MgSO4, C18, PSA and Chlorofiltr® allowed for the most sample clean up, without loss of pesticides and mycotoxins, for all cannabis samples tested. Average recovery of the 47 pesticides and five mycotoxins using the selected dSPE blend was 75.6% were as the average recovery when including GCB instead of Chlorofiltr® was 67.6%. Regardless of the sample’s original pigmentation, this blend successfully removed both chlorophyll and purple hues from all strains tested. The other six dSPE blends evaluated were unable to provide the sample clean up needed or had previously demonstrated to be detrimental to the recovery of pesticides routinely analyzed in cannabis.


References

(1)           Recommended methods for the identification and analysis of cannabis and cannabis products, United Nations Office of Drugs and Crime (2009)

(2)            W. Ross, Newsweek, (2016)

(3)            Koesukwiwat, Urairat, et al. “High Throughput Analysis of 150 Pesticides in Fruits and Vegetables Using QuEChERS and Low-Pressure Gas Chromatography Time-of-Flight Mass Spectrometry.” Journal of Chromatography A, vol. 1217, no. 43, 2010, pp. 6692–6703., doi:10.1016/j.chroma.2010.05.012.

Nevada Testing Lab Licenses Suspended, Then Reinstated

By Aaron G. Biros
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When Nevada legalized adult use sales this past summer, the market exploded and undoubtedly flooded licensed testing labs with samples to get products on shelves. In August, roughly a month after the start of adult use sales, a Las Vegas cannabis-testing lab, G3 Labs, had their license suspended for an unknown compliance issue.

“We can’t disclose the details of the suspension, including anything about penalties,” said Klapstein. “Under NRS 360.255, the information is confidential.”Then in late December, the Nevada Department of Taxation, one of the bodies tasked with regulating the state’s industry, announced in an email they suspended two more cannabis testing lab licenses. Certified Ag Lab in Sparks, Nevada and Cannex Nevada, LLC, in Las Vegas (also known as RSR Analytical Laboratories) both had their licenses suspended on December 22 and December 26 respectively.

Stephanie Klapstein, spokeswoman for the Department of Taxation, told the Reno Gazette Journal that both of those labs were not following proper protocols. “During separate, routine inspections, Department inspectors discovered that these two labs were not following proper lab procedures and good laboratory practices,” says Klapstein. “Their licenses were suspended until those deficiencies were corrected.”

According to the Reno Gazette Journal, both of those labs had their licenses reinstated and have since resumed normal business. During their license suspension, the labs were not allowed to operate and the department directed licensed cannabis businesses to submit samples to other labs. The department also directed the suspended labs in the email to coordinate with their clients who had samples in for testing; to either have their samples transferred to a different lab or a new sample taken for another lab to test. They did note that no product recalls were deemed necessary because of the suspension.

In that same email, the department directed licensed cannabis businesses to state-licensed labs in good standing, including 374 Labs, ACE Analytical Laboratory, DB Labs, Digipath Labs, MM Lab and NV CANN Lab. But on the department’s website, it says there are 11 licensed testing labs.

Back in September when we reported on the first lab license suspension, Klapstein told CIJ that under state law they couldn’t discuss any reasons behind why they suspended licenses. “We can’t disclose the details of the suspension, including anything about penalties,” said Klapstein. “Under NRS 360.255, the information is confidential.”

Because of that confidentiality, there are a number of questions left unanswered: With three lab licenses suspended in the first six months of the Nevada’s adult use market being open, how are testing labs keeping up with the market’s pace? What did those suspended labs do wrong? Do the regulations adequately protect public health and safety?

Sunrise Genetics Partners With RPC, Begins Genetic Testing in Canada

By Aaron G. Biros
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Sunrise Genetics, Inc., the parent company of Marigene and Hempgene, announced their partnership with New Brunswick Research & Productivity Council (RPC) this week, according to a press release. The company has been working in the United States for a few years now doing genomic sequencing and genetic research with headquarters based in Fort Collins, CO. This new partnership, compliant with Health Canada sample submission requirements, allows Canadian growers to submit plants for DNA extraction and genomic sequencing.

Sunrise Genetics researches different cannabis cultivars in the areas of target improvement of desired traits, accelerated breeding and expanding the knowledge base of cannabis genetics. One area they have been working on is genetic plant identification, which uses the plant’s DNA and modern genomics to create authentic, reproducible, commercial-ready strains.

Matt Gibbs, president of Sunrise Genetics, says he is very excited to get working on cannabis DNA testing in Canada. “RPC has a long track record of leadership in analytical services, especially as it relates to DNA and forensic work, giving Canadian growers their first real option to submit their plant samples for DNA extraction through proper legal channels,” says Gibbs. “The option to pursue genomic research on cannabis is now at Canadian cultivator’s fingertips.”

Canada’s massive new cannabis industry, which now has legal recreational and medical use, sales and cultivation, previously has not had many options for genetic testing. Using their genetic testing capabilities, they hope this partnership will better help Canadian cultivators easily apply genomic testing for improved plant development. “I’m looking forward to working with more Canadian cultivators and breeders; the opportunity to apply genomics to plant improvement is a win-win for customers seeking transparency about their Cannabis product and producers seeking customer retention through ‘best-in-class’ cannabis and protectable plant varieties,” says Gibbs. The partnership also ensures samples will follow the required submission process for analytical testing, but adding the service option of genetic testing so growers can find out more about their plants beyond the regular gamut of tests.

RPC is a New Brunswick provincial research organization (PRO), a research and technology organization (RTO) that offers R&D testing and technical services. With 130 scientists, engineers and technologists, RPC offers a wide variety of testing services, including air quality, analytical chemistry of cannabis, material testing and a large variety of pilot facilities for manufacturing research and development.

They have over 100 accreditations and certifications including an ISO 17025 scope from the Standards Council of Canada (SCC) and is ISO 9001:2008 certified. This genetic testing service for cannabis plants is the latest development in their repertoire of services. “This service builds on RPC’s established genetic strengths and complements the services we are currently offering the cannabis industry,” says Eric Cook, chief executive officer of RPC.

amandarigdon

Proficiency Testing in the Cannabis Industry: An Inside Look

By Cannabis Industry Journal Staff
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amandarigdon

Cannabis Labs Virtual Conference: Part 4

Proficiency Testing in the Cannabis Industry: An Inside Look
By Amanda Rigdon, Chief Technical Officer, Emerald Scientific

This presentation covers specifics of different proficiency testing schemes available to the cannabis industry. Additionally, specific challenges facing both laboratories and PT providers in the cannabis industry will be addressed. Data relating to residual solvent and potency proficiency testing will be presented.

Integrating Your LIMS System With State Tracking Systems

By Cannabis Industry Journal Staff
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Cannabis Labs Virtual Conference: Part 3

Integrating Your LIMS System With State Tracking Systems
By Hannah O’Brien, Operations Manager, Confident Cannabis

Running a lab is hard. Running a cannabis lab is harder. Watch this webinar hosted by Confident Cannabis, the most popular and only free cannabis LIMS in the country, to learn how cannabis compliance and regulatory burdens impact analytical testing laboratories in any state, and how important purpose-built software solutions are to make their business run smoothly.

Ask The Expert: Exploring Cannabis Laboratory Accreditation Part 4

By Aaron G. Biros
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In the first part of this series, we spoke with Michelle Bradac, senior accreditation officer at A2LA, to learn the basics of cannabis laboratory accreditation. In the second part, we sat down with Roger Brauninger, A2LA Biosafety Program manager, to learn why states are looking to lab accreditation in their regulations for the cannabis industry. In the third part, we heard from Michael DeGregorio, chief executive officer of Konocti Analytics, Inc., discussing method development in the cannabis testing industry and his experience with getting accredited.

In the fourth and final part of this series, we sit down with Susan Audino, Ph.D., an A2LA lead assessor and instructor, laboratory consultant and board member for the Center for Research on Environmental Medicine in Maryland. Dr. Audino will share some insights into method validation and the most technical aspects of laboratory accreditation.

Susan Audino, Ph.D.

Susan Audino obtained her Ph.D. in Chemistry with an analytical chemistry major, physical and biochemistry minor areas. She currently owns and operates a consulting firm to service chemical and biological laboratories. Susan has been studying the chemistry and applications of cannabinoids and provides scientific and technical guidance to cannabis dispensaries, testing laboratories and medical personnel. Dr. Audino’s interest most directly involves cannabis consumer safety and protection, and promotes active research towards the development of official test methods specifically for the cannabis industry, and to advocate appropriate clinical research. In addition to serving on Expert Review Panels, she is also chairing the first Cannabis Advisory Panel and working group with AOAC International, is a member of the Executive Committee of the ASTM Cannabis Section and has consulted to numerous cannabis laboratories and state regulatory bodies.

CannabisIndustryJournal: What are the some of the most significant technical issues facing an accreditation body when assessing a cannabis-testing laboratory?

Susan: From the AB perspective, there needs to be a high level of expertise to evaluate the merits and scientific soundness of laboratory-developed analytical test methods. Because there are presently no standard or consensus test methods available, laboratories are required to develop their own methods, which need to be valid. Validating methods require a rigorous series of tests and statistical analyses to ensure the correctness and reliability of the laboratory’s product, which is– the test report.

CIJ: When is method validation required and how does this differ from system suitability?

Susan: Method validation is required whenever the laboratory modifies a currently accepted consensus or standard test method, or when the laboratory develops its own method. Method validation is characterized by a series of analytical performance criteria including determinations of accuracy, precision, linearity, specification, limit of detection, and limit of quantitation. The determination of system suitability requires a series of deliberate variations of parameters to ensure the complete system, that is all instrument(s) as well as the analytical method, is maintained throughout the entire analytical process. Traditionally, method validation has been referred to as “ruggedness” and system suitability as “robustness.”

CIJ: What are the most important aspects of method validation that must be taken into account? 

Susan: In keeping with the FDA guidelines and other accepted criteria, I tend to recommend the International Conference on Harmonization (ICH), particularly Q 2A, which is a widely recognized program that discusses the pertinent characteristics of method validation. This include: method specification, linearity, range, accuracy, and precision (e.g., repeatability, intermediate precision, reproducibility). As mentioned earlier, system suitability is also a critical element and although related to method validation, does require its own protocol.

CIJ: What three areas do you see the laboratory having the hardest time with in preparing for accreditation? 

Susan: My responses to this question assume the laboratory employs appropriate instruments to perform the necessary analyses, and that the laboratory employs personnel with experience and knowledge appropriate to develop test methods and interpret test results.

  • By and large, method validation that is not appropriate to the scope of their intended work. Driving this is an overall lack of information about method validation. Oftentimes there is an assumption that multiple recoveries of CRMs constitute “validation”. While it may be one element, this only demonstrates the instrument’s suitability. My recommendation is to utilize any one of a number of good single laboratory validation protocols. Options include, but are not limited to AOAC International, American Chemical Society, ASTM, and ICH protocols.
  • Second is the lack of statistically sound sampling protocols for those laboratories that are mandated by their governing states to go to the field to sample the product from required batches. Sampling protocols needs to address the heterogeneity of the plant, defining the batch, and determining/collecting a sample of sufficient quantity that will be both large enough and representative of the population, and to provide the laboratory an adequate amount from which to sub-sample.
  • Third, sample preparation. This is somewhat intertwined with my previous point. Once an appropriate sample has been collected, preparation must be relevant to the appropriate technology and assay. It is unlikely that a laboratory can perform a single preparation that is amenable to comprehensive testing.

Steep Hill Expands To Oregon

By Aaron G. Biros
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Last week, Steep Hill announced they are expanding into Oregon with a laboratory in Portland. According to the press release, the company has licensed its testing technology to Dr. Carl Balog, a renowned pain and addiction physician.

Steep Hill has expanded significantly over the past year, including new laboratories in Pennsylvania, Maryland, Washington D.C. and Hawaii, among other states. The Berkeley-based company works in lab testing, research and development, licensing, genetics and remote testing. In 2008, Steep Hill opened the first-ever commercial cannabis-testing laboratory in the country.

Jmîchaeĺe Keller, president and chief executive officer of Steep Hill, says this is a development that will help them better understand cannabis chemistry and its medical applications. “We are pleased to announce our expansion into Oregon and especially pleased to partner with Dr. Balog, a physician who brings years of pain and addiction experience to the Steep Hill body of expertise,” says Keller. “In addition, Dr. Balog plans to use his specialized knowledge to aid Steep Hill’s research and development efforts to broaden our understanding of cannabis chemistry and to explore its wider medical applications. In partnering with Dr. Balog, we hope that Steep Hill will be able to help physicians around the United States to curb the opioid epidemic by offering Steep Hill Verified™ medicinal cannabis as an alternative to a crisis that plagues this country.”

Examination of cannabis prior to testing- credit Steep Hill Labs, Inc.

Dr. Balog, now owner and medical director of Steep Hill Oregon, says medical cannabis could be an excellent harm reduction tool, and hints at it being a possible tool in the opioid crisis. “I deal with the consequences of the opioid epidemic on a daily basis as a pain and addiction specialist,” says Dr. Balog. “The growing trend of using cannabis products as an alternative to opioids highlights the need for regulated testing. Because of the variability of marijuana preparations, testing ensures that scientific rigor is applied in a standardized way. I am dedicated to ensuring that patients have access to safe, tested cannabis, free from contaminants and to verified labels that can be trusted for their content.”

They expect Steep Hill Oregon to be open for business in the second quarter of 2018.

Ask The Expert: Exploring Cannabis Laboratory Accreditation Part 3

By Aaron G. Biros
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In the first part of this series, we spoke with Michelle Bradac, senior accreditation officer at A2LA, to learn the basics of cannabis laboratory accreditation. In the second part, we sat down with Roger Brauninger, A2LA Biosafety Program manager, to learn why states are looking to lab accreditation in their regulations for the cannabis industry.

In the third part of this series, we sit down with Michael DeGregorio, chief executive officer of Konocti Analytics, Inc., to talk method development in the cannabis testing industry and his experience with getting accredited. In the final part of this series, we are going to sit down with Susan Audino, an instructor at A2LA to learn more about the requirements where she’ll offer some advice for labs seeking accreditation.

Michael DeGregorio, chief executive officer of Konocti Analytics, Inc.

Michael DeGregorio is a doctor of pharmacy with an extensive career in medicine and scientific research. He’s worked in cancer research and medicine, teaching at the University of California, San Francisco, Yale University School of Medicine, University of Texas, Health Science Center at San Antonio and University of California, Davis. Before becoming the CEO of Konocti Analytics, a laboratory based in California, DeGregorio was also a published author in a large number of peer-reviewed medical journals.

In this piece, we sit down with DeGregorio to find out what challenges labs face when getting accredited, why they sought accreditation and their experience with getting off the ground. Stay tuned for the final part of this series!

CannabisIndustryJournal: How does a laboratory go about choosing an appropriate method in an industry where, generally, there are no validated methods available?

Michael DeGregorio: Our approach to developing analytical methods for testing cannabis began with a review of the existing laboratories and their methods, where we found no standardization and inconsistent results. Since cannabis is being used by the public and as a medicine, our goal is to help make it as contaminant-free as possible for the well-being of the consumer, and this begins by developing a state-of-the-art analytical facility.

When developing new methods, we review the published literature to see what has already been done and try to arrive at a scientifically sound consensus. We then perform experiments to determine which set of conditions works best for us. Once we have developed an appropriate method, we validate it pursuant to ISO/IEC 17025 requirements.

CIJ: How do you go about choosing what type of equipment to use for testing (e.g. by limit of detection, acceptable method use of equipment for other industries, etc.)?

Michael: After reviewing the operations of other testing laboratories, we concluded that, in general, they were not taking advantage of the most advanced technologies and had limited personnel qualified to operate it. Because public safety is our main concern, we chose state-of-the-art equipment, including GC/LC-MS with Orbitrap and ICP-MS, for testing medicinal cannabis. In addition to identifying unknown pesticides, we needed the capability of performing full chemical screening of all samples for potentially harmful compounds, e.g. steroids, present in cannabis, as well as the ability to detect trace levels of metals.

Our greatest concern is the fact that pesticides in cannabis have not been adequately studied. Current pesticide regulations suggest that government authorities believe that there are a finite number of pesticides available. Smart farmers could easily avoid the pesticides on current lists. Because of this, we chose to validate our pesticide methods with a focus on chemical classes, as opposed to specific pesticides, to give us the broadest possible coverage of potential compounds. The Orbitrap mass spectrometers also allow us to detect and identify unknown pesticides. This is something not currently being done by other laboratories. The latest microbiology methods for cannabis testing include DNA analysis, and for this we use qRT-PCR technology. Finally, the high sensitivity of ICP-MS allows for the detection of metals concentrations that may be harmful, yet undetectable by other means.

CIJ: What do you feel are the benefits of being accredited?

Michael: Being accredited shows the public that we have made a commitment to quality analytics. We feel this gives our clients peace of mind when marketing their products, knowing that they have been tested by a laboratory meeting the highest international standards of operation available using the latest technology. Furthermore, being accredited requires participation in ongoing proficiency testing programs, which helps maintain analytical competency. It should be pointed out that any prospective client of an analytical facility should take into account the laboratory’s full accredited scope of testing to ensure its competency.

CIJ: What challenges did you face during the process of getting your laboratory started and/or during the accreditation process?

Michael: Developing the quality management system and getting our equipment and processes to a state where they met accreditation requirements took several months of hard work, and turned out to be a bit more daunting than we anticipated. Our pre-accreditation assessment revealed that much work remained to be done, and it gave us a real appreciation for the level of detail and documentation required. We remained determined and eventually achieved our accreditation.

CIJ: What are the benefits to the grower and dispensaries to choosing an accredited laboratory for the testing of their product?

Michael: By choosing an accredited laboratory with a full scope of testing (potency, pesticides, mycotoxins, metals, microbiology, residual solvents and terpenes), growers and dispensaries can rest assured that their products have been tested using validated methods with appropriate quality control by trained, competent personnel. For growers, this makes their products more attractive to potential buyers. For dispensaries, this means they can confidently market their products with the knowledge that the information shown on the label is accurate, which in turn gives their customers peace of mind that the product they are consuming does not contain unacceptable levels of contaminants. 

CIJ: Why did you choose A2LA?

Michael: Once we decided to pursue accreditation, we researched the various accrediting bodies available as well as their reputations. We discovered that while all accrediting bodies are themselves accredited to the same standard, accreditation by the various bodies was not considered equal in practice. In our opinion, A2LA was considered the most prestigious, highly regarded accrediting body. Furthermore, some of the most prestigious laboratories in the country are accredited by A2LA, including Los Alamos National Laboratory, the Food and Drug Administration’s Center for Biologics Evaluation and Research, Lawrence Livermore National Laboratory, Centers for Disease Control, Federal Bureau of Investigation and the United States Department of Agriculture. Many of our preferred sources of scientific supplies and services are accredited by A2LA as well. As our goal was to be accredited by the best available accrediting body, we chose A2LA.