As the market for cannabis expands, the necessity to stay on top of industry trends and regulations becomes more and more important for cannabis industry professionals. I had the opportunity to interview Dr. Cindy Orser and draw on her invaluable cannabis science expertise.
RLS: For analytical chemists branching out into the analysis of cannabis may be a new segment of analytes and hence an attractive opportunity. California already has granted 8000 licenses for various cannabis operations. These are commercial, licensed operations that are or soon will be science-based. Science will require assays. The potential sample load should be significant.
CO: Entering the cannabis testing market today would be a lot easier than 5 years ago when I did when there were no published methods and major analytical instrument manufacturers were not yet willing to even sell their instruments into the cannabis industry. A lot has happened in 5 years; now all of the major analytical instrument makers are actively in the field and most of them have developed methods specifically for working in the cannabis matrix, which can be very challenging and harsh on columns.
RLS: I met Cindy Orser, Ph. D. at an ASTM D-37 meeting about a year ago. She attended a meeting of the stability testing Task Group in Houston, Tx. Later she co-Authored a draft practice standard that is processing through ASTM’s consensus-building gauntlet.
RLS: Please give us a brief bio including your interest in cannabis:
CO: 5½ years ago a headhunter contacted me about my interest in designing and building out a cannabis testing lab for a public company in Las Vegas – at the time I was running my own small diagnostic company in Boulder CO where we were funded by Homeland Security to develop and validate rapid diagnostics for the subunits of biological toxins. So I saw the invitation as a business opportunity to provide my expertise to start up a cannabis testing lab which I had been interested in…. Prior to my Colorado diagnostic company, ASDx Biosystems, I had spent 8 years in the DC area where I had also been recruited to start a biosensor division for a boutique physics-based defense contracting company, Arete Associates, where I chose to work on misfolded proteins but specifically, prions, with significant funding from DARPA and the NIH which in turn led to the formation of my first startup, called ADLYFE, with my DARPA project manager, Alan Rudolph. Prior to that, I had worked across many fields from remediation to molecular toxicology. I was also a tenured Professor of Biochemistry & Bacteriology at the University of Idaho following my post-doc at Purdue and PhD from the University of California at Berkeley.
RLS: Commercial labs for cannabis analytes come from two groups: Grassroots start-ups or existing labs expanding the assay menu to include cannabis assays. These face different challenges.
CO: Both approaches have their unique advantages and corresponding challenges.
RLS: There is a third segment: At-grow, even at-plant, assays to help the grower make decisions about fertilization, pesticide choice, harvest, contamination from toxins, etc.
CO: Every State is a little different in what they allow testing labs to do; but, anything beyond the prescribed testing at the State level almost always requires the grower to initiate an R&D effort with a formal request to the state.
RLS: The labs face different challenges before the first samples, but what about method development and validation?
CO: One of the biggest challenges Digipath Labs faced before ever picking up an official flower sample was convincing the State of NV to allow us to “acquire” test material for the sole purpose of method development and validation. In one instance, I actually traveled to another State to validate the Romer Labs mycotoxin assay with their participation.
The next stage is developing and validating new methods for the potential market. In some locations, Hemp for production of CBD may be the purpose of the growth. In other places, the emphasis is probably THC and THCA. Recreational cannabis also needs to consider mycotoxins, water activity, and terpenes.
RLS: How long does it take to develop and validate a method?
CO: Method development followed by validation is a time-consuming process especially when creating a first method that reflects the situation we found ourselves in 2014. Today, most instrument and assay manufacturers have developed appropriate methods for the application of their instruments/assays with cannabis samples which then requires the lab to “verify” the method and not full-blown validation.
The first step is to write down the specifications that the Method is intended to meet. After that, it is the standard investigative series:
- ACCURACY – 3 matrix samples @ 3 calibration levels with a recovery of spike at 100 +/- 30%
- PRECISION – RSD < or = to 10.0%
- SPECIFICITY – eg. For MS > 2 ions with mass accuracy < 5
- LINEARITY- r2 > or = 0.995 over a range of spiked matrix
- RANGE – min of 5 calibration points
- LOD – 3X noise level
- LOQ – 10X above background
Potency: the complicating issue in developing a method for potency is for its application across all of the different matrices from dried leaf, cured flower, extracts and a limitless variation of an edible. Initial method development for potency took several months because there were such broad variables at the beginning: e.g. What solvent? What column? How many cannabinoids to resolve?
Residual Pesticides: Nevada has a very short list of pesticides and PGRs that we are required to monitor for say in comparison with CA and OR; but basically, once a method is developed the concern comes down to what is the action level and can your instrument hit that level and in particular in extracts which are especially harsh on a triple quad.
Terpenes: Terpenes turn out to be difficult due to their varying BPs as they come off the GC column with CV acceptance levels from 50 to 150% in terpene PTs.
Microbial Contamination: For Digipath Labs, microbial options for screening are limited by the State of NV who is not up-to-date with the broad acceptance of molecular screening based on DNA and not actual plating. Under ISO 17025, we must re-validate or verify once a year, which is very time-consuming.
RLS: What about stability and use-by date?
CO: Even though stability and shelf-life testing are in the NV regulations; it not enforced. At Digipath Labs we have plans to do so. I think it should be required.
RLS: Labs generate waste. So, what do you do?
CO: YES!
Chemical – we do generate a lot of solvent waste that we collect in 55 gal drums that we contract with an environmental company to pick up once a month.
Excess Sample – referred to as RETENTION. In NV the testing labs actually go out to the grower/producers and do the sampling per State regs; which is 12 g of cured flower from a 10 lb batch; 2 g from extract derived from a 25 lb flower/trim batch.
Any material left over after testing must be kept for 30 days in a locked freezer. We are required to account for every gram of flower. After 30 days; there is an elaborate disposal process that is filmed and entails two technicians dumping the material into a 55 gal drum and updating the chain of custody inventory forms. When the drum is half full, the same environmental disposal company comes and again under camera, fills the drum with concrete and takes it to the dump.
Potential Toxins – This issue arises from the Micro Dept and has created some issues. NV requires us to use 3M PetriFilms for the enumeration of any microbial category; such as total yeast and mold and total Enterobacteriaceae which creates waste in the form of the enriched 1 g sample in a twirly bag as well as all of the PetriFilms. We routinely autoclave this waste in red autoclave bags that have the biohazard logo on them which is then thrown in the trash. The trash collectors have now refused to pick up any of the autoclaved microbial waste and have asked us to open the bags and take the 1 g enriched cannabis flower sample out of them and place it into the 55 gal drums where the retention samples end up.
RLS: What about laboratory and data integrity?
CO: At Digipath we have a Quality Policy document. It includes sections on QP-015 data integrity, ethics, and the role of technical managers.
Data integrity is the result of the processes that together assure valid data of known and documented quality. Data integrity and ethics procedures in the laboratory include training, signed and dated integrity documentation for all laboratory employees, periodic monitoring of data integrity, and documented data integrity procedures.
Data integrity training is provided for all employees initially upon hire and annually thereafter. Attendance at an initial data integrity training (part of new employee orientation) and the annual refresher training is recorded with a signature attendance sheet or other forms of documentation that demonstrates all staff have participated and understand their obligations related to data integrity.
Training records regarding data integrity and ethics are signed by each staff member annually and verified by the Lab Director or Quality Director. Topics covered are provided in writing and provided to all trainees.
RLS: What is the role of proficiency testing?
CO: PTs are an excellent exercise to participate in to check your methods by analyte category. Twice a year, Emerald Scientific orchestrates blinded proficiency testing (PT) for cannabis testing laboratories around the country. Not all states require that labs participate in PTs but it is an easy first step toward determining both where a lab stacks up in relation to their competition both in state and out of state and as preparation to undertake the process of gaining ISO 17025 accreditation.
Under Nevada regulations for medical marijuana testing labs, PTs are required to be completed twice a year for all analytes routinely tested for by the lab. PTs do not have to be those conducted through the Emerald tests across labs in the US. An independent cannabis testing lab can choose to execute an individual PT at any time with Absolute Standards, who supplies the standards used in the Emerald ring tests, where blinded samples are mailed out, the protocol is run, results submitted and then Absolute Standards will let you know how you did once the results have been reported.
The Emerald ring tests are a more accepted standardized inter-laboratory comparison PT (ILC/PT) way of providing a point of reference on how well the testing labs are performing individually and as a group. Upon a successful Emerald test, the lab is awarded a recognizable gold badge to be displayed on the lab’s website to signify a higher standard of excellence in cannabis testing. It is valid for one year for that particular Emerald Test.
RLS: Please describe DigiPath labs.
CO: Digipath Labs is in Las Vegas NV and has about 20 employees but is a public company which adds a layer of complexity in everything we do and importantly passes along all of the corporate overhead. I have always tried to be on the front edge of cannabis testing by introducing new approaches, for example developing a molecular test to distinguish hemp from marijuana versus using an arbitrary analytical value of 0.3% THC or clustering cannabis strains by their terpene content.