Tag Archives: lab

The C4 Cannabinomics Collaborative: Q&A with Dr. Zacariah Hildenbrand

By Aaron G. Biros
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Dr. Zacariah Hildenbrand, chief scientific officer and partner at C4 Laboratories, is currently researching some of the lesser-known molecules in cannabis, and he’s on to something. His research focuses on discovering new molecules, determining their therapeutic effects and expanding our understanding of the constituents of cannabis.

Dr. Zacariah Hildenbrand, chief scientific officer and partner at C4 Laboratories.
Dr. Zacariah Hildenbrand, chief scientific officer and partner at C4 Laboratories.

Dr. Hildenbrand received his Ph.D. from the University of Texas at El Paso where he researched the molecular architecture involved in hormone-dependent cancers. At the University of Texas Southwestern Medical Center in Dallas, his post-doctoral research contributed to the development of a novel therapy for the treatment of chronic myeloid leukemia, a blood-borne cancer that afflicts small children. He has published over 25 peer-reviewed scientific journal articles and hopes to do the same with his research in cannabis.

After a career of scientific consulting, Dr. Hildenbrand met Ryan Treacy, founder and chief executive officer of C4 Laboratories, in 2015 when Treacy launched the company. In June of 2015, the laboratory began operations, providing Dr. Hildenbrand the opportunity to embark on a new and exciting field of research- cannabis.

Ryan Tracy, founder and chief executive officer at C4 Labs.
Ryan Treacy, founder and chief executive officer at C4 Laboratories.

They currently collaborate with Dr. Kevin Schug of the Shimadzu Center for Advanced Analytical Chemistry (SCAAC) at the University of Texas, Arlington and together Drs. Schug and Hildenbrand are pursuing a DEA license to expand their current cannabis research. The SCAAC is a $10.0+ million analytical laboratory with instrumentation that only a handful of people in the world has access to.

C4 Laboratories, based in Mesa, Arizona, currently offers a range of services for cannabis analysis including terpene and cannabinoid analytics, microbial, pesticide, fungicide and insecticide testing. In addition to the standard gamut of tests, they also specialize in cultivation analytics like mold and mildew culture testing, viral detection with sentinel plants and comprehensive analysis of environmental conditions.

What makes their company unique is their multidisciplinary effort to characterize the therapeutic compounds found in cannabis, the C4 Cannabinomics Collaborative. We sit down with Dr. Zac Hildenbrand to talk cannabis science, his research and what they hope to accomplish with the C4 Cannabinomics Collaborative.

CannabisIndustryJournal: What is the C4 Cannabinomics Collaborative?

Dr. Zacariah Hildenbrand: The C4 Cannabinomics Collaborative is an open collaboration between growers and scientists to discover new molecules in cannabis and to have a better characterization of individual cannabis strains based on the active constituents found in each sample. We are facilitating the collaboration of some of the world’s best cannabis growers with world-class scientists to find new information about the plant.

What we want to accomplish in this work is identifying novel molecules. Because of the [federal government’s] restrictions in researching cannabis, there is very little peer-reviewed literature on many of the compounds found in cannabis. We want to secondarily find out what those molecules do in the human body and thus make recommendations for strains targeting specific conditions.c4 logo

We also want to understand the strains currently out there by determining the most established cannabinoids and terpenes via chemotyping. You hear a lot of people talking about the effects of an Indica or Sativa and making recommendations based on that. We want to find chemical signatures based on cannabinoids and terpenes and make recommendations based on that. There are a lot of problems at hand when discussing strain names scientifically. There are nomenclature issues- people calling the same strain different names, people giving multiple names to the same strain to make it appear that their strain portfolios are more diverse.

We can identify the chemical signatures in strains based on the major cannabinoids and terpenes. Based on the terpenes and chemical profile we can determine more accurate recommendations for patients as well as in recreational applications. All of this, again, discovering the new molecules, identifying the current strains, is so we can make more informed decisions regarding cannabis use. It is not a panacea but it is a very robust plant. There are a lot of terpenes with anti-inflammatory responses. Other molecules help with blood flow, sleep, regulating blood glucose, and we all know the cases of CBD helping children with convulsions and epilepsy. We want people to make sure they have the most up-to-date information.

CIJ: How is your collaboration with the SCAAC at UT Arlington contributing to this work?

Dr. Hildenbrand: One of the instruments we use there is a supercritical-fluid-extraction supercritical-fluid-chromatography mass-spectrometer (SFE-SFC-MS). With that instrument, we can do the extraction on the machine with an extreme level of sensitivity. It is ideal for drug discovery and identifying molecules in the parts-per-quadrillion range. This particular instrument allows us to detect molecules with an extreme level of sensitivity without volatizing them during the sample extraction process.

The Shimadzu Center for Advanced Analytical Chemistry
The Shimadzu Center for Advanced Analytical Chemistry

We want to acquire samples of unique cannabis from growers that will work with us to discover new cannabis constituents. We are in the process of getting a DEA license so that we can send products across state lines to the center at UT Arlington to perform the advanced characterization. They have instrumentation that only a handful of people in the world have access to, which gives us the best opportunity to explore the unknown. When we discover new molecules, find out what they do on the molecular level, we can then isolate these compounds and ultimately use this newfound knowledge for the development of effective nutraceuticals.

CIJ: What molecules are you researching right now?

Dr. Hildenbrand: Some of the low-hanging fruit in our research looks at identifying compounds similar to the better-studied compounds such as THC and CBD. THCV has a very similar structure to THC, but has a shorter acyl carbon chain (3 carbons vs. 5).

Tetrahydrocannabivarin (THCV)
Tetrahydrocannabivarin (THCV)

THCV doesn’t induce a psychoactive response (like THC), but it does improve fat utilization, so it has remarkable potential for medicine. We are looking at what conditions are required for it to occur naturally. Cannabis doesn’t produce THCV in a high amount. 0.7% by weight is the most we have seen in Arizona. In Oregon, where craft cannabis has been refined to a much higher degree, we have heard rumblings of some strains containing up to 3% THCV. We want to find out if this is a possible weight loss tool. Our research in CBDV is very much the same.Cannabinoid Biogenesis

CBL is the breakdown product of CBC when it is treated with ultraviolet light. We know absolutely nothing about what CBL does. If we find a strain that produces high amounts of CBC, we can then treat it with UV light and force the conversion to CBL, and then ultimately determine what it does. This is a good example of low-hanging fruit and the versatility of cannabis. Based on the biogenesis of the cannabinoids, we can alter the profile of cannabis products using a series of biochemical reactions.

Cannabicyclol (CBL)
Cannabicyclol (CBL)

For example, we have been helping clients in Arizona look for a quality sleep aid in cannabis. Certainly, Indica strains will help, but the molecule CBN helps specifically with sleep abnormalities. As CBN is formed as a byproduct when CBD or THC are oxidized, we see some producers using liquid nitrogen to oxidize CBD, leading to higher CBN levels. I would like to think we are in the age of understanding CBD, THC and the major terpenes,but there are a whole milieu of compounds that require our attention and THCV, CBDV and CBL are just a few that we want to devote our efforts to right away.

CIJ: What are your plans in the immediate future?

Dr. Hildenbrand: We are in the process of finalizing the documents to bring a C4 laboratory into Oregon where we can do quite a bit of research and where we’ll have access to some very unique cannabis. We will offer full compliance testing per ORELAP and OLCC regulations, but we also want to acquire samples (free of charge) from growers that want to collaborate with us to discover new molecules. We’ve been lucky enough to start working with growers like Adam Jacques and Chris West in Eugene, but we also want to be available to other growers who want to contribute to this research.

CIJ: What are your long-term goals with this project?

Dr. Hildenbrand: At a basic level, we hope to expand the current understanding of the cannabis plant. There is a lot of “bro science” and anecdotal claims out there. There is so much that we don’t know about cannabis that we cannot simply rely on anecdotal claims for each strain. We want to bring cannabis into the same light as any pharmaceutical-grade or biomedical research.

We need to be characterizing this plant with the same level of detail as other pertinent molecular therapies. In doing so there are a lot of potential discoveries to be made and we might be able to unlock the future of medicine. A drug like Marinol, for example, has been met with mixed reviews because its only one dimensional. Furthermore, we find that the terpene molecules are tremendously beneficial and this interplay between cannabinoids and terpenes is something that we want to explore further. All and all we wish to further illustrate the therapeutic capacities of cannabis within the contexts of specific ailments and medical conditions, while discovering the medicine of the future.

Second Oregon Health Alert for Tainted Cannabis with Pesticides.

By Aaron G. Biros
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Three health alerts were issued Thursday, November 4th for contaminated cannabis sold to consumers at North Bend, Salem and Eugene dispensaries. Green-Way Medicinal in Salem and Stonies in North Bend sold two strains of cannabis flower found to have high levels of piperonyl butoxide, an ingredient commonly found in pesticides that acts as a synergist to amplify the effects of certain compounds.

The two batches in question, including the strains Pleeze (batch number G6J0039-02) and Dryzl (batch number G6J0039-01), were found to contain the potentially dangerous chemical at levels of 15.39 ppm and 16.24 ppm, respectively. The Oregon Health Authority (OHA) action level for piperonyl butoxide is 2.0 ppm. To see the full health alert, click here.

The dispensary in Eugene, Flowr of Lyfe, sold one strain of cannabis that had levels of the insecticide spinosad over the 0.2-ppm action level. The very popular indica hybrid, Dutch Treat (batch number G6J0018-01), was found to contain 0.9-ppm of spinosad. Though it still tested above the 0.2-ppm action level for that insecticide, it pales in comparison to October’s health alert, where a batch of cannabis had over 200 times the acceptable level of that insecticide. Both spinosad and piperonyl butoxide are considered toxic to humans.oha_logo_lrg

According to the health alert, “All tests were performed by an OHA-accredited and Oregon Liquor Control Commission-licensed laboratory.” It is unclear exactly how or why the cannabis was able to get transported and transferred from the grower to the dispensary and then sold to consumers after failing the pesticide test. According to Jonathan Modie, spokesman for the OHA, they are currently investigating the matter and following up with the dispensaries and growers to find out what happened. “We need to find out how this got transferred in the first place and then sold,” says Modie. “They had access to the test results and should have been able to determine for themselves that these products should not have been sold or transferred.”

“We don’t know, we are still gathering information, there is a risk of civil penalty as well as losing your registration for a dispensary or grower that illegally transferred products that have tested for analytes above the action levels,” says Modie, when asked if punitive measures would be taken. While there are no particular regulations for this scenario in performing a mandatory recall, the OHA is obligated under law to issue health alerts when there is a situation that might affect public health, according to Modie.

“We deal with this with infectious disease outbreaks or during a food borne illness outbreak; if they [the public] can avoid it by hearing from us then we want to get the word out and this is a very similar situation.” For medical patients that purchase potentially contaminated cannabis such as this, it is easy to contact them to have the patient dispose or return the cannabis. Dispensaries are not required to collect information from recreational customers, and most dispensaries do not, which is a major problem when this situation happens, as it has twice in the past two weeks.

“We can never do too much communication,” says Modie. “We will let the public know in any way possible that they should return this product or dispose of it responsibly.”

The Practical Chemist

Building the Foundation of Medical Cannabis Testing – Understanding the Use of Standards and Reference Materials – Part 2

By Joe Konschnik
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In the last article I referred to the analogy of the analytical reference material being a keystone of the laboratory foundation, the stone upon which all data relies. I then described the types of reference materials and their use in analytical testing in general terms. This article will describe the steps required to properly manufacture and deliver a certified reference material (CRM) along with the necessary documentation.

A CRM is an exclusive reference material that meets strict criteria defined by ISO Guide 34 and ISO/IEC 17025.  ISO is the International Organization for Standardization and IEC is the International Electrotechnical Commission. These organizations work together to set globally recognized standards. In order for a reference material to be labeled as a CRM it must 1) be made with raw or starting materials which are characterized using qualified methods and instruments, 2) be produced in an ISO-accredited lab under documented procedures, and 3) fall under the manufacturer’s scopes of accreditation. Verifying a CRM supplier has these credentials is easily done by viewing their certificates which should include their scopes of accreditation. Restek_accredit

There are many steps required to produce a CRM that meets the above three criteria.  The first step requires a review of the customer’s, or end-user’s requirements to carefully define what is to be tested, at what levels and which analytical workflow will be used.  Such information enables the producer to identify the proper compounds and solvents required to properly formulate the requested CRM.

The next step requires sourcing and acquiring the raw, or starting materials, then verifying their compatibility and stability using stability and shipping studies in accordance with ISO requirements. Next the chemical identify and purity of the raw materials must be characterized using one or more analytical techniques such as: GC-FID, HPLC, GC-ECD, GC-MS, LC-MS, refractive index and melting point. In some cases, the percent purity is changed by the producer when their testing verifies it’s different from the supplier label. All steps are of course documented.

restek_CRMThe producer’s analytical balances must be verified using NIST traceable weights and calibrated annually by an accredited third party provider to guarantee accurate measurement. CRMs must be prepared using Class A volumetric glassware, and all ampules and vials used in preparation and final packaging must be chemically treated to prevent compound degradation during storage. Next, CRMs are packaged in an appropriate container, labeled then properly stored to maintain the quality and stability until it’s ready to be shipped. All labels must include critical storage, safety and shelf life information to meet federal requirements. The label information must be properly linked to documentation commonly referred to as a certificate of analysis (COA) which describes all of the above steps and verifies the traceability and uncertainty of all measurements for each compound contained in the CRM. Restek_CRM2

My company, RESTEK, offers a variety of documentation choices to accompany each CRM. Depending on the intended use and data quality objectives specified by the end-user, which were defined way back at the first step, three options are typically offered: They include gravimetric only, qualitative which includes gravimetric, and fully quantitative which includes all three levels of documentation. The graphic to the right summarizes the three options and what they include.

It’s important to understand which level you’re purchasing especially when ordering a custom CRM from a supplier. Most stock CRMs include all three levels of documentation, but it’s important to be sure.

Understanding what must be done to produce and deliver a CRM sets it apart from other reference material types, however it’s important to understand there are some instances where CRMs are either not available, nor required and in those situations other types of reference materials are perfectly acceptable.

If you have any questions or would like more details about reference materials please contact me, Joe Konschnik at (800) 356-1688 ext. 2002 by phone, or email me at joe.konschnik@restek.com.

Oregon Issues Health Alert for Contaminated Cannabis

By Aaron G. Biros
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According to Jonathan Modie, spokesman for the Oregon Health Authority (OHA), on Friday, October 21st, the OHA issued a ‘health alert’ regarding cannabis products sold from a McMinnville dispensary that were possibly tainted with extremely high levels of Spinosad, an insecticide commonly used to combat mites and other pests. “My understanding is that two medical patients purchased the cannabis products whom we had contact info for, but most of the purchasers were recreational customers,” says Modie. “Because it is not required to get contact info for recreational customers, we issued the health alert to get the word out as quickly as possible because we didn’t know who bought the product.” The OHA is urging consumers who purchased cannabis from New Leaf CannaCenter in McMinnville to check the labels and see if they purchased potentially dangerous cannabis, and to either return the cannabis to the dispensary or dispose of it appropriately.

oha_logo_lrgThe action level, the measured amount of pesticides in a product that the OHA deems potentially dangerous, for Spinosad is 0.2 parts-per-million (PPM). The two batches in question are the strains Dr. Jack (batch number G6J0051-02) and Marion Berry (batch number G6J0051-01), which were tested to contain approximately 42 PPM and 22 PPM respectively, much higher than the OHA’s action level.

While this is the first health alert issued in Oregon in connection with potentially contaminated cannabis, Modie says he expects there will be more health alerts in the future. “Unfortunately the product was inappropriately transferred from the grower to the dispensary and from the dispensary to customers, so we are working to get the word out to dispensaries, growers and processors about the testing rules to prevent this from happening in the future,” says Modie. “We want to make it clear that any grower, processor or dispensary that does not follow the testing requirements or fail to label, store or retain batches that fail a test will be subject to enforcement actions such as fines, penalties, suspension or revocation of their license.” The OHA has a list of pesticide analytes and their action levels on their website.

“We are advising recreational and medical users alike to read the product labels closely; the labels must have the license or registrant number, the packaging or distributor license number, the name of the strain and the universal symbol,” says Modie. “We are also suggesting consumers request a copy of pesticide test results from the dispensary.” It is unclear at this time if all of the cannabis products in question have been properly disposed of, but OHA was informed that New Leaf has pulled all products in question off of the shelf.

Steep Hill Labs Expands to Pennsylvania, Washington, D.C.

By Aaron G. Biros
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Last week, Steep Hill Labs, Inc. announced plans to expand on the East Coast, including licensing for laboratories in Washington, D.C. and Pennsylvania. The cannabis testing company now is operating or developing in seven states, the District of Columbia along with an official arrangement with a research university in Jamaica, according to Cathie Bennett Warner, director of public relations at Steep Hill.

The same team of physicians that oversees the Steep Hill laboratory in Maryland will operate the Pennsylvania and D.C. labs. Heading that team is chief executive officer Dr. Andrew Rosenstein, chief of the division of Gastroenterology at University of Maryland Saint Joseph Medical Center and assistant clinical professor of Gastroenterology and Hepatology at the University of Maryland Medical Center. Dr. Rosenstein has been recognized by Baltimore Magazine as a top doctor in the Baltimore area, according to a press release.

Dr. Andrew Rosenstein, CEO of Steep Hill Maryland, PA and D.C.
Dr. Andrew Rosenstein, CEO of Steep Hill Maryland, PA and D.C.

According to Dr. Rosenstein, they want to provide accurate clinical results for trials with patients using cannabis. “All clinical trials will require a competent, credible and reliable lab partner and that is what we are bringing to the field- and that is why we are working with Steep Hill,” says Dr. Rosenstein. With team members having backgrounds in pathology, molecular diagnostics, clinical chemistry, microbiology and genetics, it should come as no surprise that they plan to participate in clinical research.

Dr. Rosenstein’s vested interest in cannabis safety stems from prior experience with his patients using cannabis. “Over the past five years, we have seen an increased number of patients using cannabis, particularly for managing the side effects of Crohn’s disease and cancer treatment,” says Dr. Rosenstein. “They would bring it up to us and at the time I didn’t know much about it, but anecdotally it’s really clear that a lot of patients have great responses to it.” Not knowing much about the preparation or safety of cannabis at the time led Dr. Rosenstein to advise patients to be very careful if they are immunocompromised.

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

“When a patient is immunocompromised, a bacterial or fungal infection can be lethal, so because we had patients using cannabis, we wanted to make sure it was safe,” says Dr. Rosenstein. So when Maryland legalized medical cannabis, Dr. Rosenstein and his team saw the need to protect patient safety and Steep Hill was a perfect fit. “We really didn’t want to reinvent the wheel so we looked for someone to partner with,” says Dr. Rosenstein. “Steep Hill has the best technology and the best credibility and we didn’t want to compromise on quality and safety issues. They felt the same way so we partnered with them and culturally it has been a great fit.”

Steep Hill Express in Berkeley, CA- MD,PA and D.C. will have a similar offering of instant potency analysis
Steep Hill Express in Berkeley, CA- MD,PA and D.C. will have a similar offering of instant potency analysis

The new laboratories plan to offer a similar range of services that are offered at other Steep Hill labs, such as rapid potency testing for THC-A, ∆-9-THC, CBD, CBD-A and moisture. But Dr. Rosenstein sees clinical opportunities in the East Coast medical hubs. “We want to provide the testing component for studies, providing clinical reproducibility and consistency, and those are the things as a top-notch lab that we are interested in doing.”

A petri dish of mold growth from tested cannabis- Photo credit: Steep Hill-
A petri dish of mold growth from tested cannabis- Photo credit: Steep Hill Labs, Inc.

With a physician-led group that has experience in molecular diagnostics, partnering with Steep Hill is about being medically focused, according to Dr. Rosenstein. “First and foremost, this is about patient safety.” Because of that, he emphasizes the need for required microbiological contaminant testing, particularly because of his experience with patients. “If you’re a cancer patient and you get a toxic dose of salmonella or E. coli, that can kill you, so testing for microbiologic  contamination is of the highest priority.”

According to Warner, bridging the medical cannabis science gap with Steep Hill’s professionalism and experienced doctors practicing medicine is a big deal. “We are working very closely with their medical team to make sure these standards are medically superior,” says Warner. “To have these doctors with such a high level of knowledge in medicine working with us in cannabis analytics is a breakthrough.”

The Nerd Perspective

Detecting the Undetectable

By Amanda Rigdon
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In my last column, I took a refreshing step out of the weeds of the specifics behind cannabis analyses and took a broader, less technical look at the cannabis industry. I had envisioned The Nerd Perspective being filled with profound insights that I have had in the cannabis industry, but I have realized that if I restricted this column to insights most would consider profound…well…there would not be many articles. So in this article, I want to share an insight with you, but not one that is earth shattering. Instead, I want to talk about a simple concept in a way that might help you think a little differently about the results your lab generates, the results you have to pay for or even the results printed on a cannabis product you might purchase.

This article is all about the simple concept of concentration – the expression of how much of something there is in relation to something else. We use expressions of concentration all the time – calories per serving, percent alcohol in beer, even poll results in the presidential election circus. Cannabis is not excluded from our flippant use of concentration terms – percent cannabinoid content, parts-per-million (ppm) residual solvents, and parts-per-billion (ppb) pesticides. Most of us know the definition of percent, ppm, and ppb, and we use these terms all the time when discussing cannabis analytical methods. During my career in analytical chemistry, it has occurred to me that parts per billion is a really infinitesimal amount…I know that intellectually, but I have never tried to actually visualize it. So being the nerd that I am, I went about comparing these often-used concentration terms visually in my kitchen.

I started by preparing a 1% solution of food coloring paste in water. This was accomplished by weighing out 5g of the food coloring and dissolving it into 500mL of water (about one teaspoon into a pint). The resulting solution was so dark it was almost black:

rsz_percent2

The picture above expresses the low end of what we care about in terms of cannabinoid concentration and a pretty normal value for a high-concentration terpene in cannabis.

I then took one teaspoon of that mixture and dissolved it into 1.32 gallons of water (5mL into 5000mL), resulting in a 10ppm solution of green food coloring in water:

rsz_ppm

I did not expect the resulting solution to be so light colored given the almost-black starting solution, but I did dilute the solution one thousand times. To put this into perspective, 10ppm is well above many state regulatory levels for benzene in a cannabis concentrate.

I then took one teaspoon of the almost-colorless 10ppm solution and dissolved that into another 1.32 gallons of water, resulting in a very boring-looking 10ppb solution of green food coloring in water:

rsz_1ppb

Obviously, since I diluted the almost-colorless 10ppm solution a thousand times, the green food coloring cannot be seen in the picture above. As a reference, 10ppb is on the low end of some regulations for pesticides in food matrices, including – possibly – cannabis. I know the above picture is not really very compelling, so let’s think in terms of mass. The picture above shows eleven pounds of water. That eleven pounds of water contains 50 micrograms of food coloring…the weight of a single grain of sand.

To expand on the mass idea, let’s take a look at the total mass of cannabis sold legally in Colorado in 2015 – all 251,469 pounds of it. To express just how staggeringly small the figure of 10ppb is, if we assume that all of that cannabis was contaminated with 10ppb of abamectin, the total mass of abamectin in that huge amount of cannabis would be just 1.143g – less than half the mass of a penny.

To me, that is an extremely compelling picture. The fact is there are instruments available that can measure such infinitesimal concentrations. What’s more, these tiny concentrations can be measured in the presence of relatively massive amounts of other compounds – cannabinoids, terpenes, sugars, fats – that are always present in any given cannabis sample. The point I’d like to make is that the accurate measurement of trace amounts of cannabis contaminants including pesticides and residual solvents is an astounding feat that borders on magical. This feat is not magic though. It requires extremely delicate instrumentation, ultra-pure reagents, expert analysts, and labor-intensive sample preparation. It is far from trivial, and unlike magic, requires a large investment on the part of the laboratories performing this feat of science. Other industries have embraced this reality, and the cannabis industry is well on its way toward that end…hopefully this article will help put the job of the cannabis analytical lab into perspective.

Automated Solutions for Cannabis Laboratories: Part I

By Danielle Mackowsky
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rsz_96_well_plate-1-1
Using well plates for dSPE sorbents can help expedite sample clean up.

Sample volume remains to be the primary influence on whether an automated solution is a logical investment for a cannabis testing facility. Due to both the complexity of the material being tested and the extraction approach at hand, it may be difficult to find an automated platform that can fully accommodate your laboratory’s needs. Hamilton Robotics in collaboration with United Chemical Technologies (UCT) has developed a solution that allows for automation of specific sample clean up steps commonly utilized in cannabis pesticide testing schemes. The MPE2 Positive Pressure Extraction/Evaporation Module is a standalone manifold that can also be incorporated into a number of automated liquid handling decks. Used in tandem with dispersive solid phase extraction (dSPE) salts/sorbents packed into a 96 well plate, this combination provides laboratories with high throughput extraction convenience with comparable results to traditional dSPE for the analysis of over forty pesticides.

As states continue to expand testing requirements for pesticides, it is vital that your laboratory is equipped with a method that allows versatility for the addition of new compounds without burdening your extraction team. There are a variety of dSPE salt and sorbent blends readily available that have been optimized for cannabis extractions. This allows for the use of a reliable extraction technique that can be adapted for the automation age. Hamilton is widely recognized throughout both clinical and forensic laboratory settings and the MPE2 platform is an excellent first system for laboratories beginning to automate/semi-automate their processes.

MPE2 Positive Pressure Extraction/Evaporation Module
MPE2 Positive Pressure Extraction/Evaporation Module

Following an initial QuEChERS extraction, additional cleanup is typically recommended for extracts that are being analyzed for pesticide content due to the low detection limits often required. dSPE provides the necessary sample clean up to obtain those thresholds, but often burdens a laboratory staff with additional time consuming preparation steps. Traditionally, dSPE salts are packed into 2 mL centrifugation tubes that require a cumbersome supernatant pipetting step followed by additional vortex, spin and transfer steps. By packing the dSPE sorbents into a well plate format, the user is able to completely automate this above described clean up ultimately saving time and adding convenience without jeopardizing any recovery data.

For most compounds, the recovery was greater than 65% for both methods of dSPE. The mean recoveries for traditional dSPE were 98.0%, 99.2% and 97.9% at pesticide concentrations of 50 ng/mL, 100 ng/mL and 200 ng/mL, respectively. For comparison, the mean recoveries at the same concentrations for well plate dSPE were 85.0%, 88.9% and 89.1%. Therefore, there was typically about a 10-11% absolute difference in recovery between the two methods, which can be corrected for by implementing the use of internal standards. When comparing the recovery differences between the two methods, there are six compounds with noticeably larger discrepancies across all three concentrations, namely: chlorpyrifos, cyprodinil, diazinon, spinetoram, spiromesifen 278 and trifloxystrobin. If these data sets are excluded, then the average absolute differences in recovery between the two methods decrease to 8.8%, 6.4% and 5.8% for concentrations of 50 ng/mL, 100 ng/mL and 200 ng/mL, respectively.rsz_1shutterstock_226135945-1

Overall, laboratories can estimate on saving 40-60 minutes per 96 samples processed using the Hamilton MPE2 in conjunction with a UCT dSPE plate. When a liquid handling robot is also available, this time saving estimation is potentially doubled. Time spent per sample, including the training of laboratory scientists, is an important factor to consider when setting up your laboratory. Automation is in an investment that can greatly reduce a laboratory’s overall labor costs in the long run.

Oregon Cannabis Lab Accreditation Program Gets Help, Problems Addressed

By Aaron G. Biros
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Last week, news of problems facing Oregon’s cannabis laboratory accreditation program surfaced, leading some to speculate about possible delays for the recreational cannabis market. According to The Register-Guard, ORELAP administrator Gary Ward believed the program was “on the precipice of collapse.”

oha_logo_lrgAccording to Jonathan Modie, spokesman for the Oregon Health Authority (OHA), the Oregon Environmental Laboratory Accreditation Program (ORELAP) was anticipating over 30 cannabis laboratories applying for accreditation and they doubled their staff from two to four to prepare for the uptick in applications.

In June, the agency had zero labs applying for accreditation but within two months, 37 labs applied. However, the Oregon Department of Environmental Quality (DEQ) just provided three additional staff members on Monday to help with the application process, says Modie.

Some believe the issues could mean the state may not have enough accredited labs by October 1st, when the recreational cannabis market is expected to go into full swing. “It is difficult to say exactly how many labs we can accredit by October 1,” says Modie. “We have seven labs today which would bring it to nine labs waiting for assessment, but our goal is to get as many labs assessed and hopefully accredited as soon as possible.”

With the additional staff members, Modie is hopeful this will jumpstart the program. “We really appreciate our collaboration with the DEQ and look forward to boosting our capacity a bit to help us get through this busy time,” says Modie.

Part of the reason some laboratories might have trouble meeting prerequisites is simply because the requirements are very strict. “The process involves submitting a quality manual, standard operating procedures, method validation, submitting proficiency testing data and finally undergoing an ORELAP assessment by our staff, so it is a very rigorous process,” says Modie. “This speaks to our concern for making sure they have the right systems in place so public health is protected.” Modie said there were at least three labs that did not pass the assessment.

Roger Voelker
Roger Voelker, lab director at OG Analytical

Bethany Sherman, chief executive officer of OG Analytical, believes the hardest part of the process involves getting accredited for testing pesticides. OG Analytical, based in Eugene, Oregon, has already received their accreditation, one of the first to do so. “The pesticide testing requires our most expensive instrumentation and the sample preparation for testing pesticides is the most time consuming,” says Sherman. “Not only does it require very specific instrumentation, it also requires a real know-how and expertise to ensure we are cleaning samples appropriately, minimizing background noise and looking at the pesticides in trace quantities.” According to Sherman, laboratories are also left to their own devices to develop methodologies specifically for the cannabis matrix, adding to the difficulties.

Rodger Voelker, Ph.D., lab director at OG Analytical, seems confident that the state will be able to handle it. “It is a relief they were able to get some resources from the DEQ and I think the state will not allow a program with this kind of importance to fall apart,” says Voelker. He believes after this initial phase of putting the program in place, the workload will go down. “It is easier to maintain a program than it is to implement,“ adds Voelker. In his eyes, it is crucial for the program to require rigorous science. “People are forced to reconcile that there is a tremendous amount of controls to be considered to produce legally defensible data and I think it is great that the requirements are so strict.”

The OHA’s job is to essentially safeguard public health and they do not want to leave any stone unturned when it comes to potential contamination, says Modie. “This is not just about getting as many labs accredited as possible, this is about protecting public health.”

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The Nerd Perspective

‘Instant’ Cannabis Potency Testing: Different Approaches from Different Manufacturers

By Amanda Rigdon
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This is the first piece of a regular column that CIJ has been so kind to allow me to write for their publication. Some readers might recognize my name from The Practical Chemist column in this publication. Since the inception of that column, I’ve finally taken the plunge into the cannabis industry as chief technical officer of Emerald Scientific. Unlike The Practical Chemist, I will not spend the entire first article introducing the column. The concept is simple: while I find the textbook-esque content of The Practical Chemist scintillating, I have a feeling that the content is a little too heavy to spring on someone who is looking for engaging articles over their precious coffee break. Instead, The Nerd Perspective will consist of less-technical writing focusing on my experience and insights for the cannabis industry as a whole. But don’t worry – I’m sure I will not be able to refrain from technical jargon altogether.

To kick off the column, I want to talk about instrumentation for ‘instant’ cannabis potency testing. At this point, it’s common knowledge in the cannabis analytics industry that the most accurate way to test cannabis potency is through extraction then analysis by HPLC-UV. I agree wholeheartedly with that sentiment, but HPLC analyses have one drawback: they can be either inexpensive or fast – not both. There are some instruments entering the market now that– while not as directly quantitative as HPLC-UV – promise to solve the inexpensive/fast conundrum. During my most recent trip to California, I was able to spend some quality time with two well-known instrument manufacturers: SRI Instruments and PerkinElmer, both of whom manufacture instruments that perform fast, inexpensive cannabis potency analyses. From my previous home at the heights of The Ivory Tower of Chromatography: Home of the Application Chemists, SRI and PE couldn’t be more different. But as seen through the eyes of a company who deals with a wide range of customers and analytical needs, it turns out that SRI and PE are much the same – not only in their open and honest support of the cannabis industry, but also in terms of their love of all things technical.

My first stop was SRI Instruments. They are a relatively small company located in an unassuming building in Torrance, CA. Only a few people work in that location, and I spent my time with Hugh Goldsmith (chief executive officer) and Greg Benedict (tech service guru). I have worked with these guys for a few years now, and since the beginning, I have lovingly referred to them as the MacGyvers of chromatography. Anyone familiar with SRI GCs knows that what they lack in aesthetics, they make up for in practicality – these instruments truly reflect Hugh and Greg’s character (that’s meant as a compliment).

SRI specializes in relatively inexpensive portable and semi-portable instruments that are easy to set up, easy to operate, and most importantly – engineered for a purpose. It’s actually really hard to manufacture an instrument that meets all three of these criteria, and the folks at SRI accomplish this with their passionate and unique approach to problem solving. What I love about these guys is that for them, nothing is impossible. Here’s an example: the price of the portable GC-FID instruments SRI builds is inflated because the instruments require separate – and pricey – hydrogen generators. That’s a big problem – hydrogen generators are all pretty much the same, and none of them are cheap. This didn’t faze SRI: they just decided to design their own super small on-board hydrogen generator capable of supplying hydrogen to a simple GC macgyversystem for six hours with just 20mL of distilled water from the grocery store! I’m not kidding – I saw it in action on their new Model 420 GC (more on that in some future pieces). Was the final product pretty? Not in the least. Did it work? Absolutely. This kind of MacGyver-esque problem solving can only be done successfully with a deep understanding of the core principles behind the problem. What’s more, in order to engineer instruments like these, SRI has to have mastery over the core principles of not only chromatographic separation, but also of software development, electrical engineering, and mechanical engineering – just to name a few. These quirky, unassuming guys are smart. SRI is a company that’s been unapologetically true to themselves for decades; they’ll never be a contender for beauty queen, but they get the job done.

On the surface, PerkinElmer (PE) contrasts with SRI in almost every way possible. With revenue measured in billions of dollars and employees numbering in the thousands, PE is a behemoth that plays not only in the analytical chemistry industry but also in clinical diagnostics and other large industries. Where SRI instruments have a characteristic look of familiar homeliness, PE instruments are sleek and sexy. However, PerkinElmer and SRI are more alike than it would seem; just like the no-frills SRI, the hyper-technical PE instruments are engineered for a purpose by teams of very smart, passionate people.

DoogieWith its modest price tag and manual sample introduction, the SRI Model 420 is engineered for lower throughput users to be a fast, simple, and inexpensive approach to semi-quantitative process control. The purpose of the instruments manufactured by PE is to produce the highest-quality quantitative results as quickly as possible for high-throughput labs. PE instruments are built using the best technology available in order to eke out every last ounce of quantitative accuracy and throughput possible. Fancy technology is rarely inexpensive, and neither is rigorous product development that can last years in some cases. In a way, PE is Doogie Howser to SRI’s MacGyver. Like MacGyver, Doogie is super smart, and his setting is a sterile hospital rather than a warzone.

I had a wonderful conversation with Tim Ruppel, PE’s headspace-GC specialist, on the sample introduction technology incorporated into the TurboMatrix Headspace Sampler, where I also learned that the basic technology for all PerkinElmer headspace-GC instruments was designed by the men who wrote The Book on headspace gas chromatography: Bruno Kolb and Leslie Ettre**. Later, I was able to get a much-needed lesson on FT-IR and the Spectrum Two IR Spectrometer from Brian Smith, PE’s spectroscopy expert, who actually wrote the book on quantitative spectroscopy***. Tim and Brian’s excitement over their technology mirrored that of Hugh and Greg. It turns out that SRI and PerkinElmer are more alike than I thought.

These two instrument manufacturers have addressed the fast/inexpensive conundrum of cannabis potency testing in two different ways: SRI’s instrument is extremely inexpensive, easy to operate, and will provide semi-quantitative values for THC, CBD, and CBN in just a few minutes; PE’s instrument is more expensive up front, but provides quantitative (though not directly quantitative) values for all of the major cannabinoids almost instantly, and requires almost no maintenance or consumables. These two instruments were designed for specific uses: one for inexpensive, easy use, and the other for more comprehensive results with a higher initial investment. The question consumers have to ask themselves is “Who do I need to solve my problem?” For some, the answer will be MacGyver, and for others, Doogie Howser will provide the solution – after all, both are heroes.


** B. Kolb, L. Ettre, Static Headspace-Gas Chromatography: Theory and Practice, John Wiley & Sons, Hoboken, NJ, 2006.

*** Brian C. Smith, Quantitative Spectroscopy: Theory and Practice, Elsevier, Boston, MA, 2002.

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Quality From Canada

The Devil is in the Detail – Changes to Canada’s Cannabis Regs to Encourage Patient Independence and Business Competition

By Tegan Adams, Elfi Daniel-Ivad MSc
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Canada’s new ACMPR was launched late last month on August 24th. The key change that most notice is that Canadians may now again grow their own cannabis at home for medical purposes. In addition, more strict guidelines for product testing and labeling requirements for Licensed Producers (LPs) were released.health-canada-logo

Short term pain for long term gain. While the combination of allowing patients to grow at home and more strict regulations for LPs may at first seem like a business disadvantage; overtime LPs will be thankful for the combination switch. Health Canada’s new requirements encourage a leveling of the playing field globally between LPs and large scale product manufacturers of pharmaceuticals, therapeutics and natural health products. The steps Health Canada is taking to regulate our producers, is exactly what they need to get ready for mass production that will be necessary for recreational markets, scheduled for release in Spring 2017.

Picture rows of Tylenol bottles on the shelf at your favorite pharmacy. Now picture rows of cannabis bottles on the shelf beside them. This is what medical cannabis will look like in Canada perhaps as early as 2018, if not sooner. With just under forty LPs on the map and a projected sales volume of modest billions, Canada’s LPs’ eyes are widening with dollar signs as they lube up their oil production and more to see what shelves in Canada will hold.

Curious to know more? Our regulatory department manager Elfi Daniel-Ivad is an expert in regulatory change. She has worked on close to 150 submissions for cannabis licensees in Canada and beyond. Here are a few key changes from her department’s overview to better understand.

MMPR ACMPR (Updated)
No personal production or designated production available to patients (aside from that grandfathered in by MMAR). Personal production and designated production available. Patients may grow 5 indoor plants OR 2 outdoor plants at any given time per gram prescribed to them.
Licensed Producers were not required to label THC or CBD amounts in dried cannabis, though most producers did for sales and educational purposes. Oils had to be labeled with THC and CBD amounts. Licensed Producers must label their percent THC and CBD for dried and fresh cannabis products.
For the labelling of oils, the total quantity of THC, CBD and oil in a container had to be shown. Restrictions on THC allowed no more than 10mg/mL THC per capsule and no more than 30mg/mL THC per mL oil to be distributed. In addition, oil labels must now include information on “carrier” oil and allergen information. Containers must be labelled with number of capsules, the net weight and volume of each capsule. .
No reference to validation of analytical testing methods. Analytical testing must be completed using validated testing methods; confirming reliability and consistency in results for   contaminants, disintegration, residues and THC, THC-A, CBD and CBD-A
Accredited labs can only test products as received from Licensed Producers. In addition to Licensed Producers, patients growing their own or having a designated grower growing for them may also test their products at an accredited lab.

In addition to these changes, it is important to note that if an individual or company has an MMPR proposal already submitted they can now revise it to include oil production (previously, it was first dried bud only). If a company submits a new ACMPR proposal, they can include oil production on their application right away. Interested in submitting your own application? Or need help with one in the USA? Our regulatory department would be happy to answer any questions you might have about the process.