Rapid Screening of Cannabinoids in Edibles Using Thermal Desorption-GC–MS

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CANNABIS SCIENCE AND TECHNOLOGY | VOL 1 • NO 4 www.CannabisScienceTech.com

CANNABIS ANALYSIS 50 Rapid Screening 12 Error, Accuracy, of Cannabinoids in and Precision 30 Edibles Using Thermal Brian C. Smith Desorption-GC–MS Find out if you know the difference Rojin Belganeh and William Pipkin between accuracy and precision 30 Leveraging Selectivity A look at how TD-GC–MS analysis and Effi ciency to Take eliminates conventional sample prep- CULTIVATION CLASSROOM the Strain Out of LC–UV aration regimes and can be used as a Method Development for good rapid screening technique. 18 Curbing the Cannabinoid Profi ling Cannabis Industry’s Justin Steimling Appetite for Energy Here the LC–UV separation of 16 can- Roger Kern nabinoids of interest was performed A look at energy consumption in the while the potential impact from minor 56 cannabis industry and suggestions on cannabinoids and terpenes on report- how cultivator’s can reduce those costs. ed potency values was monitored. 56 Major Engine of FEATURES 36 Certifi ed Reference Growth in the Cannabis Material Manufacturing Industry: State of the Art 22 Setting the Standard: Challenges Formulation Technology Considerations When Don Shelly for Product Innovation and Handling DEA-Exempt What to consider when creating or us- Expanded Applications Cannabinoid Reference ing certifi ed reference materials Andreas M. Papas Standard Preparations Used Which formulation choices will sup- for Potency Determination TUTORIALS port safety, effi cacy, stability, and Catharine E. Layton consumer acceptance and Andrew J. Aubin Reference standard qualifi cations are CANNABIS CROSSROADS discussed and techniques for qualify- 40 ing incoming reference standards are 60 Professor Jack Henion suggested. Shares Insights on Cannabis 40 Holding Data to a Research, Academia, and Higher Standard, the Expanding Role of Part II: When Every Peak Mass Spectrometry Counts—A Practical Guide Joshua Crossney to Reducing Contamination Professor Jack Henion shares his in- and Eliminating Error in sights on cannabis research, includ- the Analytical Laboratory ing challenges and opportunities in Patricia Atkins moving cannabis science forward. In this guide, we look at the most common sources of contamination DEPARTMENTS and error in an analytical process. 10 Caannnabis NNewss Foocuus

on the cover: 62 Apppplicationn Nootes Canna Obscura/Shutterstock.com 63 Suuppplier Prrofi lees

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-&"11"-1BSUT $POTVNBCMFTt"UMBOUJD"WF 3BMFJHI /$ttXXXQBMQBSUTDPN 8 Editorial Addvisoory Board

EDITORIAL ADVISORY BOARD MEMBERS

Susan Audino, S.A.Audino & Associates, LLC Sandy Mangan, SPEX SamplePrep LLC Bob Clifford, Shimadzu Scientifi c Instruments David (Dedi) Meiri, Laboratory of Cancer Ashlee Gerardi, Restek Corporation Biology and Cannabinoid Research, Technion Israel Institute of Technology Jacklyn Green, Agate Biosciences Kevin Schug, Department of Chemistry & Zac Hildenbrand, Inform Environmental, LLC Biochemistry, The University of Texas at Arlington Karan Kapoor, Avana Canada Inc. Brian Smith, Big Sur Scientifi c Autumn Karcey, Cultivo, Inc. Katherine Stenerson, MilliporeSigma Julie Kowalski, Trace Analytics

Cannabis Science and Technology’s Editorial Advisory Board is a group of distinguished individuals assembled to help the publication fulfi ll its editorial mission to educate the legal cannabis industry about the science and technology of analytical testing and quality control. With recognized expertise in a wide range of areas, board members perform various functions, such as suggesting authors and topics for coverage, reviewing manuscripts, and providing the editor with general direction and feedback. We are indebted to these individuals for their contributions to the publication and to the cannabis community as a whole.

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Cannabis Science and Technology The course will be called “Horticulture of Cannabis: Announces Editorial Advisory Board From Seed to Harvest” and will be taught by Gerald “Gerry” Berkowitz, a professor of plant science, and ex- Cannabis Science and Technology is pleased to an- perts working in the cannabis industry today such as nounce the launch of its editorial advisory board (EAB). CEOs from successful business operations, for exam- The distinguished members of the EAB will play a cru- ple, licensed cannabis growing facilities in Connecticut cial role in defi ning the editorial direction and maintain- and cannabis testing laboratories. ing a high level of editorial quality and integrity. According to UConn Today, the course will have a The offi cial charter of the advisory board for Canna- fi rm focus in horticulture, serving as a gateway to the bis Science and Technology is to support the publica- entire cannabis industry. The introductory course will tion in identifying the most signifi cant developments be open to all UConn students, regardless of their main the cannabis science fi eld, and in ensuring the rele- jor, and has no prerequisites for enrollment. In addition vance and technical accuracy of our content. In doing to Berkowitz, expert guest lecturers will present on top- this work, board members support our mission to edu- ics such as cannabis genetics, seed selection, soil and cate the legal cannabis industry about the science and tissue testing, plant hormones, and laboratory testing technology of analytical testing and quality control. We of harvests. are pleased to announce the following board members: Berkowitz currently has the ability to work with low • Susan Audino, S.A. Audino & Associates, LLC tetrahydrocannabinol (THC) hemp cannabis plants, • Bob Clifford, Shimadzu Scientifi c Instruments which will be used as examples for class demonstra- • Ashlee Gerardi, Restek Corporation tions. As reported by UConn Today, the plants will be • Jacklyn Green, Agate Biosciences grown in concert with the course to provide students • Zac Hildenbrand, Inform Environmental, LLC with hands-on examples of the range of horticultur- • Karan Kapoor, Avana Canada Inc. al methods applied to cannabis, such as propagating • Autumn Karcey, Cultivo, Inc. clones through cuttings, transplanting, training plants • Julie Kowalski, Trace Analytics to alter canopy and fl ower architecture, culling males, • Sandy Mangan, SPEX SamplePrep LLC pest management, and more. The course will be of- • David (Dedi) Meiri, Laboratory of Cancer Biology fered in the Spring 2019 semester (2). and Cannabinoid Research, Technion Israel Institute of Technology References • Kevin Schug, Department of Chemistry & 1) https://today.uconn.edu/2018/10/cannabis- Biochemistry, The University of Texas at Arlington course-responds-industry-need/#. • Brian Smith, Big Sur Scientifi c 2) http://www.plantscience.uconn.edu/. • Katherine Stenerson, MilliporeSigma

For more information about our esteemed board Cannabis Science Conference members, please see the full announcement on our Announces East Coast Show website: www.cannabissciencetech.com/news/cannabis-science-and-technology-announces-editorial-advisory-board. The organizers of the Cannabis Science Conference announced a new east coast show for 2019. Canna- University of Connecticut to bis Science Conference East will take place April 8–10, Offer Cannabis Horticulture Course 2019, at the Baltimore Convention Center in Baltimore, Maryland. The fl agship conference in Portland, Ore- gon will take place September 4–6, 2019. The University of Connecticut (UConn, Storrs, Connect- Oral and poster abstracts are now being accept- icut) announced plans to offer a class on cannabis hor- ed for Cannabis Science Conference East. Please vis- ticulture through the Department of Plant Science and it www.cannabisscienceconference.com for more infor- Landscape Architecture (1). mation on both events. TOTEMART/SHUTTERSTOCK.COM

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Error, Accuracy, and Precision In the science of analytical chemistry, we quantitate things such as weights and concentrations. All of these measurements generate a number, but to truly understand the quality of data we have to know the size of the error in the measurement. Well known measures of data quality include accuracy and precision. Do you know the difference between these two metrics? Do you know how to quantitate them? And how does all of this apply to cannabis analysis? Please read on to fi nd out more.

Brian C. Smith

ny time a quantity is meas- voice is the signal and the static in the presence of unwanted atmospher- ured, be it your weight on a connection is the noise. If the volume ic carbon dioxide inside the instru- bathroom scale, the speed of the caller’s voice is large compared ment. If you see a CO peak in an FT- A 2 of light, or potency of a cannabis to the static, the connection has a IR spectrum that you measure, it is bud, there will be error involved in good SNR and you can clearly hear an artifact. the measurement. Sources of er- what the other person is saying. Al- A measurement of a signal-to- ror include environmental changes, ternatively, if the static in the connec- noise ratio, of course, needs a signal. power fl uctuations, electronics, and tion is high, and the caller’s voice can In chromatography, spectroscopy, good old fashioned human error. Er- barely be heard above it, the SNR of and other analytical techniques the ror exists because human beings are the call is low and you will have trou- signal is the magnitude of the meas- not gods and thus cannot control all ble understanding what your caller is urement made. This is illustrated in the variables all the time for any giv- saying. Note that a high SNR phone Figure 2. en measurement (1). Two of the most call, or any high SNR data, will carry a The y-axis in Figure 2 is in absorb- important types of noise are random lot of information. Whereas a low SNR ance units (AU), which is a meas- noise and systematic noise. Let’s dis- phone call or low SNR data will carry ure of the amount of light absorbed cuss random noise fi rst. very little information. This is why SNR by a sample. The size of the peak is a measure of data quality. at 461 cm-1 in this case is the signal, Random Noise In analytical chemistry error is of- which has a magnitude of 0.0215 AU. Random error is caused by varia- ten seen as “fuzz” in the baseline of A measure of the error in a signal is bles we cannot control as mentioned chromatographic and spectroscopic called the peak-to-peak noise (PPN). above. The sign of random error is measurements. An example of this is In Figure 2, this is calculated by tak- random, that is, it is equally probable seen in Figure 1. ing a section of the baseline and tak- to be positive or negative. This is why Figure 1 shows noise measured by ing the highest noise point, in this measurements are often times ex- a Fourier transform-infrared (FT-IR) case 0.01388 AU, subtracting from pressed as X±y, where X represents spectrometer. Since the sign of ran- it the lowest noise point, which is the value of the measurement and dom noise is random, the baseline 0.001144 AU, and obtaining a PPN of y represents the amount of error in fl uctuates up and down randomly. 0.00244 AU. The signal-to-noise ratio the measurement. Error is sometimes The size of these wiggles is a measure is then (0.0215) AU/(0.00244) AU for a called noise, and the quality of data of the noise (2). Note in Figure 1 that value of ~9. This is not a particularly can be expressed as a signal-to-noise the size of the noise varies with wave- good SNR. Many laboratories meas- ratio (SNR) defi ned in equation 1. number, which is typical of any spec- ure what they call a limit of detection trum measured using light (electro- (LOD), which is typically calculated SNR = (Signal)/(Noise) [1] magnetic radiation). as 3x the noise in a measurement or The big peak at 2350 cm-1 in Fig- an SNR of 3. By calculating an LOD a The SNR concept is perhaps best il- ure 1 is an artifact, which is a peak laboratory is saying, “this is the min- lustrated using a cell phone call. In or signal in your data that is not from imum signal I can reliably measure.”

this case, the volume of the caller’s the sample. This peak is from the Many cannabis laboratory pesticide BLABLO101/SHUTTERSTOCK.COM

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the better SNR. Random error cannot be eliminated, but it can be min-

100.4 imized by controlling error sources and averaging data. 100.2

100 Systematic Error Systematic error occurs when a 99.8 measurement is consistently wrong % Transmittance

99.6 by the same amount and in the same direction. A classic example of sys- 99.4 tematic error is a clock that has been 3500 3000 2500 2000 1500 1000 500

Wavenumber (cm-1) not been set ahead for daylight savings time. This clock will always be Figure 1: An example of noise in an infrared spectrum, seen as the random wiggles or 1 h behind whenever you look at it. “fuzz” in the baseline. It is an example of systematic error because it is always off by the same amount, 1 h and in the same direc-

.022 tion, always behind.

.02 To detect systematic error, one Peak-to-peak noise @ must have a true or reference value 440–420 cm-1 .018 Peak height (0.01388 – 0.01144) = with which to compare your obser- (signal) = 0.0215 0.00244 .016 @461 cm-1 vation. In the United States the ref-

Absorbance erence time value is supplied by the .014 atomic clock at the National Institute .012 of Standards and Technology (NIST)

500 480 460 440 420 Wavenumber (cm-1) in Gaithersburg, Maryland (3). Unlike random error, systematic error can be

Figure 2: Measurement of a signal-to-noise ratio. In this case, the size of the peak is the eliminated, in this case by simply set- signal and the size of the random fl uctuations in the baseline are the noise. ting the clock forward 1 h.

Accuracy and Precision analysis reports, for example, con- above the limit of detection but be- Accuracy and precision are terms in tain an LOD that gives the minimum low the limit of quantitation. common use, and they are common- amount of a pesticide whose pres- Random noise can be reduced by ly confused. Some people seem to ence can be reliably confi rmed. observing the same quantity multiple think they are the same thing. They However, signals at or just above times and then averaging those read- are not. Precision is a measure of the the LOD are too noisy to give relia- ings. The SNR improves as the square scatter in a set of measurements. Ac- ble quantitative results. So, many lab- root of the number of observations, curacy is a measure of how far away a oratories use a limit of quantitation N, averaged as such: set of measurements is from the true (LOQ) of 10x the noise level or an SNR values. This is illustrated in Figure 3. of 10. By calculating an LOQ a labo- SNR ∝ (N)1/2 [2] Imagine you are tasked with weigh- ratory is saying, “this is the minimum ing a 1.0 g standard weight on the amount of signal with which I can give where N is the number of observa- same scale seven times. Because of a reliable quantitative measure.” The tions averaged. error you will not get the same read- LOQ in a pesticide report from a can- In essence, equation 2 works being each time, but a spread of val- nabis analysis laboratory tells you the cause random noise cancels itself as ues. This is illustrated to the left in minimum amount of pesticide that N increases. Equation 2 is why we pre- Figure 3. The bullseye represents can be reliably quantitated in a sam- fer to measure an average instead of the known value, 1.0 g, and the sev-

ple. Thus, the SNR of 9 in Figure 2 is a single observation, the average has en dots represent the seven readings BLABLO101/SHUTTERSTOCK.COM

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you obtained. These readings are a value. This is the ideal situation. Accu- let’s say you lose your 1.0 g standard random scatter and they are not par- racy is a better measure of data quali- weight, repeated measurements on ticularly close to the bullseye; this ty than precision since it incorporates some other sample can be made to data set is imprecise because the both random and systematic error. obtain a precision value. For exam- readings are widely scattered and Now, if one does not have a true ple, you might want take a pencil and not reproducible. This data is inaccu- or reference value to compare to, measure its weight seven times. The rate because of how far on average the readings are from the bullseye, the true value. If your seven readings are about the same value, let’s say 1.9 g, they would form the plot seen DON’T LET QUALITY in the middle of Figure 3. The readings form a tight cluster in the up- GO TO THE DOGS per right hand corner of the bullseye, but their center is far from the true value of 1.0 in the center. This dataset is said to be precise but inaccurate. It is precise because the scatter in the data is small and the points cluster tightly together. Precision is a measure of the amount of random error in a dataset. It is found by measur- ing the same value on the same sample multiple times. Imprecise data will give a wide scatter, like to the left in Figure 3. Precise data will be tightly clustered as seen in the middle of Figure 3. However, this data is clustered around 1.9 g, far away from the true value of 1.0 at the center of the bullseye. Thus, this data set is inaccu- FEED YOUR EQUIPMENT H2, N2 rate because the points fall far from the true value. Precision is a useful AND ZERO AIR ON-DEMAND measure of data quality since it quan- titates random error, but it ignores • Consistent Purity systematic error so it is not a com- • Consistent Pressure plete picture. The systematic error in • Proven Safe the middle of Figure 3 is the distance between the center of the data clus- • Cost Effective ter and the bullseye. • Eliminates Cylinder Storage Accuracy is a measure of the and Delivery Issues amount of random and systematic error in a data set. If the seven weights Visit us on-line or call for a consult you measure all cluster tightly around 1.0 g, the diagram at the right in Figure with one of our sales engineers today. 3 will be obtained. This data is precise because the spread of the readings is +1-203-949-8697 small, and it is accurate because the points cluster tightly around the true www.ProtonOnSite.com

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analysis. Because of the lack of this guidance, laboratories have had to fend for themselves to develop their own methods. Thus, different laboratories have developed different ways of doing things, and this leads to different results. In particular, I have observed that many laboratories prepare samples in completely different ways, lead- Imprecise and Precise and Precise and ing to variations in the results obtained. inaccurate inaccurate accurate All of this is not to say that cannabis laboratories are

Figure 3: An illustration of the concepts of accuracy and preci- not precise, they are. They can get impressively repro- sion using bullseyes. ducible data on a given sample. But until SRMs and standard methods are available, the search for a true ac- spread of these values will give you the precision of the curacy and a solution to the inter-laboratory variation is- balance. This is not as good as an accuracy determina- sue will, I believe, be problematic. In the meantime, it tion, but since we do not have a standard weight, and we might be best for cannabis businesses to fi nd a laborato- do not know the true weight of the pencil, the precision ry they trust, submit their samples to them, and only com- is the best we can do. When reporting error size in the pare numbers from this chosen laboratory to themselves literature, always make sure to disclose whether you are going forward. reporting an accuracy or a precision, and for an accuracy result always clearly state the source of the true value References used in the calculations. 1) B.C. Smith, Quantitative Spectroscopy: Theory and Practice, (Elsevier, Boston, Massachusetts, 2002). Is True Accuracy Achievable in the 2) B.C. Smith, Fundamentals of Fourier Transform Infrared Cannabis Industry Right Now? Spectroscopy, (CRC Press, Boca Raton, Florida, 2011). As mentioned above, obtaining an accuracy calculation 3) www.time.gov. requires a sample with a known value to be measured. For many industries, where plant material is processed 4) www.nist.gov. such as tea, there exist NIST traceable standard refer- 5) M.O. Bonn-Miller, M.J.E. Lofl in, B.F. Thomas, ence materials (SRMs). For example, there exist SRMs for J.P. Marcu, T. Hyke, and R. Vandrey, JAMA, tea and tea extracts that can be used to give reference J. Am. Med. Assoc. 318, 1708 (2017). values for accuracy calculations (4). Since cannabis is not 6) B. Smith, P. Lessard, and R. Pearson, legal at the federal level in the United States, NIST, being manuscript in preparation. a Federal agency, cannot develop cannabis SRMs for any of the materials that cannabis laboratories test. This has left the industry in the diffi cult position of having to develop its own standards and analytical methods. Now, for chromatographic methods there are pure cannabinoid standards that can be used for calibration. However, since there are no SRMs available for the actu- About the Columnist al types of samples analyzed, such as cannabis buds and extracts, the effect of sample preparation on the fi nal Brian C. Smith, PhD, results is not refl ected in the accuracy calculations per- is Founder, CEO, and Chief Technical Officer of Big Sur Scientific in Capitola, California. Dr. Smith formed. Thus, I have to conclude, perhaps controversial- has more than 40 years of experience as an in- ly, that in the absence of the proper SRMs, true accura- dustrial analytical chemist having worked for such cy measurements in the cannabis analysis industry may companies as Xerox, IBM, Waters Associates, and Princeton Instruments. For 20 years he ran Spectros not be possible. Associates, an analytical chemistry training and consulting This issue is refl ected in the problem of inter-laborato- firm where he taught thousands of people around the world ry variation. This has been documented before and con- how to improve their chemical analyses. Dr. Smith has written three books on infrared spectroscopy, and earned his PhD in tinues to be an issue (5,6). In my view, part of the prob- physical chemistry from Dartmouth College.

lem is the lack of industry standard methods for cannabis BLABLO101/SHUTTERSTOCK.COM

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18 Cultivationn Claassroom

Curbing the Cannabis Industry’s Appetite for Energy

Cannabis cultivation has a problem, a big problem: the electric bill! Earlier in my cannabis career, I heard growers complain about the cost of electricity eating into their profi ts—but now the situation is much worse, while the stakes are far greater. Energy consumption is under greater scrutiny than ever by regulators because of concerns over fossil fuels hastening climate change and the cost of expanding the grid. This means the cannabis industry better fi nd solutions to cut down on its power use, or else the government will—and no one wants that.

Roger Kern

ust how bad is the energy problem in the cannabis in- Light emitting diodes (LEDs) are fi nding their way into more dustry? Energy inputs are estimated by several stud- and more grows, despite skepticism by cultivators. Most of Jies to be perhaps as much as half of total indoor can- the controversy centers around suffi cient power during the nabis cultivation costs. Based on this estimate, let’s assume fl owering phase and the nonsolar nature of the spectrum. 2000 kW/h per pound of medicine and a typical energy cost Both concerns are not issues in the greenhouse, where LEDs of $0.24 (1). For growing a single pound of cannabis, that’s serve to augment the solar spectrum during off-peak times of $480 worth of energy! And just what is this energy used for? solar illumination such as early morning, late afternoon, winter It’s not just lighting; also sucking up power are air conditioners months, and cloudy days. However, the energy benefi ts are real and chillers, dehumidifi ers, carbon and high-effi ciency partic- and substantial: Most manufacturers claim that their 600-W ulate air (HEPA) fi lters, fans, and system controllers. In a 2016 fi xture can replace a 1000-W high-intensity fi xture, resulting study done for investor-owned San Diego Gas & Electric, Ev- in a 40% reduction in lighting costs. This effi ciency is further ergreen Economics broke down the energy consumption for compounded by energy savings associated with cooling be- indoor cannabis production and found that lighting accounts cause of the heat produced from lighting; therefore, these for 38% of the energy consumed, venting 30%, and air condi- practices are being adopted by CEA vegetable growers on a tioning 21% (2). Furthermore, when one considers the current global scale. The reason is simple: LEDs save money by saving means of energy production, the cultivation of 1 kg of can- energy in the CEA setting while increasing yield and quality. nabis results in the production of 4600 kg of carbon dioxide. As the acceptance of cannabis spreads across the nation Clearly, there is plenty of room for improvement and, just and world, it’s logical to assume more will be grown, which, as fortunately, there are many ways to improve. For now, let’s of course, means an increasing demand for electricity. The in- just talk about two: the cultivation structure and how it is il- dustry would be wise to demonstrate its responsibility by act- luminated. According to BOTEC Analysis, indoor cultivation ing now to cut back on power consumption. Because if cul- has the highest energy consumption levels with 4400 kWh, tivators don’t, strangers may be more than willing to act for 6100 KWh/kg. Greenhouses have much lower energy de- them—and when they do, they may not have the industry’s mands with 6 kWh, 580 kWh/kg (3). (BOTEC’s study consid- wellbeing in mind. ers the energy consumption of outdoor cultivation to be minimal.) This is why I recommend to cultivators that they start An Expert’s Point of View their site and structure selection process by giving serious When looking for solutions to the cannabis industry’s high en- consideration to a greenhouse. ergy consumption, it’s necessary to have experts who are fa- In California, my home state, the Department of Agricul- miliar with both cultivation and how utilities can help grow- ture has projected the following trends: There will be a shift in ers. Dan Duran has that knowledge, having worked for power production from the northern to the southern part of the state companies and, as a professor, having researched cultivators’ while the use of controlled-environment agriculture (CEA) is use of power. Dan spoke with Cannabis Science and Technol- forecast to jump dramatically over 2–3 years from 16% to more ogy magazine to share how the two sides can work together. than 50% (4). I expect there will always be indoor, greenhouse, and outdoor production, and there are advantages to all these What has been your background and experience with methods. Still, I would like to see the percentage of greenhouse the cannabis sector? facilities increase for the reduction of energy use while still ena- Dan Duran: I am a veteran of 25 years in the corporate world, bling a controlled environment for maximum yield and quality. where I led operations and marketing teams in the IT and

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energy sectors, including stints with American Telephone spoken to several of the utilities and the large consulting fi rms and Telegraph, Nippon Electric Corp., Southern California that help them with strategic planning and program evalu- Edison, and Sempra Energy. I decided, after the tragic events ation, and while there are a few fi eld people who recognize on September 11, to move into academia and have been a the need to learn and support growers, the senior manage- faculty member at a California liberal arts college since then. ment teams have almost consistently chosen to take limited At Whittier College, I study domestic and international sus- initiatives. This is not a good situation, as the growing num- tainable development with a focus on energy and renewable ber and size of CEA and greenhouse growers need to better resource management. I founded Energized Solutions, LLC, understand and take advantage of “Time of Use rates,” en- in 2002 with the goal to provide real-world data and research ergy-effi ciency programs and incentives, service planning in- pertaining to sustainable development and renewable re- frastructure support, technical training on current and inno- source management to be used in and out of the classroom. vative technologies, and a broad spectrum of other services Dr. Cinzia Fissore, who joined Energized Solutions a few years and support provided to the traditional medium to big users. ago, and I have planned and executed several fi eld-based Utilities increasingly want to be perceived as being environ- research projects pertaining to agriculture for the Califor- mentally responsible with strong corporate social responsibil- nia Public Utilities Commission (CPUC) and California-based ity agendas, but in this case, they have largely missed the op- electric, natural gas, and water utilities. We have conduct- portunity to acknowledge the emergence of this new base of ed more than 200 fi eld visits with a broad spectrum of grow- customers that are producing a high-value crop with signifi - ers, from row crops to dairy, nuts, and vineyards. Our goal is cant social and economic benefi ts. to better understand current practices, help the utilities and other stakeholders learn more about their customers, and What should current and hopeful growers expect from develop and deliver programs to improve energy–water effi - the utilities and what is the most effective approach to ciency and other sustainability-focused programs and tech- use in securing technical and fi nancial support from them? nical services. Over the last few years, we have tightened our Duran: Utilities are typically organized by function, such as focus on the urban agriculture sector and more specifi cally distribution, energy–water effi ciency, service planning, cus- on CEA and greenhouse operations, especially the growth in tomer education, regulatory, and public relations. My fi rst Southern California. piece of advice is to do a bit of internet leg work and determine where the agriculture support team is situated and to You mentioned your energy–water work with utilities contact the local account executive or service representative. and the agriculture sector. What are the utilities While most of the utility agriculture teams are located in the thinking and doing with respect to the legalized and traditional rural locales, they are the most likely to respond fast-growing cannabis business in California, and to a request for information on services. Growers should also specifi cally with the increasing focus on CEA and identify the local or regional public affairs or equivalent rep- greenhouse production by growers? resentative because they are the most attuned to listening to Duran: The good news is that a few utilities have recognized users and their professional association advocates. The pub- the need and opportunity to work with CEA and greenhouse lic affairs folks are charged to be the face of the utility to the growers, including those in urban areas. The case is that the local community and the local power structure, and they are majority of the utilities have opted to take a “wait and see” responsible for articulating problems and opportunities to position. As a veteran manager of several utilities, the reality their utility management organization. While most of the mid- is that utilities are far more risk-adverse than other fi rms in the to larger utilities run education and training programs on ef- private or public arena, and while most of them highlight their fi ciency strategies and new product offerings, they have yet commitment to serving customer needs, their primary goal to develop and offer programs customized to the needs of is to protect investors by heeding to the dictates and regu- the CEA or greenhouse cannabis grower. With the excep- lations of the CPUC or equivalent oversight agency. For ex- tion of the recent 2017 Grower Report prepared by the Can- ample, while the major utilities in California have dedicated nabis Conservancy for the Colorado Energy Offi ce (5), there agriculture account executives or service representatives that is a lack of solid and current data on the energy spends and work with the large growers and post-harvest processors that effi ciency levels of different energy-intensive (for example, have signifi cant energy and water needs, the unfortunate case lighting, heating, ventilation, and air conditioning [HVAC], and is that they rarely, if ever, interact, and know very little about CO2 generation) and water-operating modes used for out- the needs of CEA and greenhouse growers. Another factor is door and indoor operations. Still, it is commonly stated and that many of those growers have operated in a “gray” area to believed that the energy spend constitutes between 30–45% elude the detection by utilities and most often take the “fast” of CEA-greenhouse grower operations. There is a real lack of in–out operations way rather than a systems approach. I have understanding regarding which specifi c energy-use areas can

VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 www.CannabisScienceTech.com | CANNABIS SCIENCE AND TECHNOLOGY 20 Cultivationn Claassroom

be best addressed via more energy-effi cient technologies cultivators have a strong commitment to the environment, and what their impact is on the bottom line. while still making a profi t. This approach will generate a positive relationship with the utilities, create goodwill in the pub- What should utilities put on their agenda to prepare and lic eye, and demonstrate that cannabis cultivation is on the support the CEA-greenhouse growers? cutting edge of sustainability. By being innovative in cultiva- Duran: The cannabis growers market has several key stake- tion practices and becoming leaders in lowering energy us- holders, and the local energy, water, and waste utilities all age, cannabis cultivators will show the world that they are ex- have important roles to play in helping this industry mature cellent stewards of our natural resources and good partners with respect to energy effi ciency, water management, and with public utilities. waste practices. At the moment, I am in conversation with several Southern California utilities about how best to prepare References and support the CEA and greenhouse growers. The most im- 1) J.P. Caulkins, “Estimated Cost of Production for Legalized portant step is for the utilities to prepare and execute a data- Cannabis” (RAND Drug Policy Research Center, 2012). gathering plan that identifi es and records the current ener- 2) Evergreen Economics, “SDG&E Cannabis gy, water, and waste practices—what we call a baseline study. Agriculture Energy Demand Study” (Prepared for Utilities need to fi gure out where the near-future CEA-green- San Diego Gas & Electric Company, 2016). house growth will be with respect to their distribution sys- 3) M. O’Hare, D. Sanchez, and P. Alstone, “Environmental tems, including both the electric and water distribution grids. Risks and Opportunities in Cannabis Cultivation” (BOTEC Electric utilities tend to be “grid-focused,” and it is critical that Analysis Corporation internal publication, June 2013). they understand where the electric load will grow and how to 4) J. Remillard and N. Collins, “Trends and Observations of best support it. The corollary need is for utilities to identify and Energy Use in the Cannabis Industry” (ACEEE Summer work with the growers to provide a spectrum of services and Study on Energy Effi ciency in Industry, 2017). support, from infrastructure service planning to information 5) Colorado Energy Offi ce, “Energy Use in the on how demand charges are calculated, the available time of Colorado Cannabis Industry” (Fall 2018). use and other rate options, and the incentives and rebates for current and innovative technologies, including LED and virtu- al management systems. I believe that utilities will be motivat- ABOUT THE INTERVIEWEE ed to respond to the growers who can provide data on plant- Dr. Dan Duran cycle growing needs, energy consumption by equipment and is an Associate Professor at Whittier College where he teaches sustainable development. He is also the system, and operations practices. My hope is that the sustain- founder and principal of Energized Solutions, LLC, a ably and profi t-focused CEA sector and greenhouse growers CPUC-certified minority-owned business (MBE). His will work with the utilities, equipment and tech providers, and career includes senior planning and operations work with energy utilities and water agencies. His ongoing research consultants to plan and implement a triple bottom line strate- and consulting focus is in the energy–water nexus, especially with gy that generates value and simultaneously benefi ts people, the California agriculture community. He has met with hundreds the planet, and profi ts. It is my impression that utilities are very of agriculture producers to identify their sustainability and best practices, and he works with utilities to optimize the potential for motivated to work with other sectors to promote leadership CEA cannabis production. in effi ciency and design (LEED) buildings and facilities that are both energy effi cient and operationally advanced. My “call” ABOUT THE GUEST COLUMNIST to growers is to work with their local utilities (gas, electric, water, and waste), equipment and technology providers, and ex- Dr. Roger Kern perienced consultants to lay the foundation for LEED growing is a scientist and technologist who cares deeply about the cultivation and health of plants in the operations that produce quality products using environmen- cannabis industry. With his PhD in microbiology tally-responsible growing operations that increase bottom- from the University of California, Davis, Plant line profi ts and generate value for both consumers and the Growth Laboratory, he solves the most challenging problems in hydroponics, from studying the root microbi- cities and counties in which they operate. ome to developing nutrients and lighting systems to ensure plant health and a disease-free lifecycle. He spent 22 years at Conclusion NASA’s Jet Propulsion Laboratory as a scientist, technologist, and research leader before becoming the President of Agate We are entering a new era of cooperation and coordination Biosciences, a consulting firm for project management, sys- between the cannabis industry and the public utilities. Re- tems engineering, and science in CEA for the past eight years. ducing energy consumption and adopting more sustainable He leads developments to optimize sustainability, consistency, quality, and yield without compromising plant health. practices should become a high priority for cultivators. This Direct correspondence to: [email protected] will show the public and the utility companies that cannabis

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Setting the Standard: Considerations When Handling DEA-Exempt Cannabinoid Reference Standard Preparations Used for Potency Determination

With the growth of the cannabis market, signifi cant investments have been made into the setup, operation, and maintenance of cannabinoid potency testing laboratories. Results generated in these laboratories are not only dependent upon the accuracy of the testing methods and sample preparation, but on the quality of the reference standards from which batches of cannabis samples are directly quantifi ed. The quality, consistency, and reliability of reference standard solutions are not only a priority for manufacturers, but it is in the best interest of testing laboratories to protect and maintain the integrity of these formulations during use and storage. In this manuscript, reference standard qualifi cations are discussed and techniques for qualifying incoming reference standards are suggested. Additionally, in-laboratory handling of reference solutions is explored to promote and preserve accuracy of the formulations during use.

Catharine E. Layton and Andrew J. Aubin

urrently in the United States, A reference standard is broad- Certifi ed cannabis testing labo- cannabinoid isolates are clas- ly described by the U.S. Food and ratories operate under Internation- Csifi ed as Schedule I controlled Drug Administration (FDA) and U.S. al Organization for Standardization substances under the Controlled Pharmacopeia (USP) as a well char- (ISO) and International Electrotechni- Substance Act (CSA) Title 21 Code of acterized, highly purified com- cal Commission (IEC) 17025 accredi- Federal Regulations (CFR) Parts 1300 pound that serves as a specimen or tations issued by the American Asso- to 1308. The distribution of these iso- performance calibrator from which ciation for Laboratory Accreditation lates is controlled and monitored by drug substances, excipients, im- (A2LA) to provide confi dence that re- the Drug Enforcement Administra- purities, degradation products, or sults generated in these testing lab- tion (DEA) division of the U.S. De- compendial reagents are identified oratories are high quality and con- partment of Justice (1). Purchasing and quantified for purity and po- sistent. Reference standards used and use of controlled substances tency. Any material can be called a by these laboratories are held to the must adhere to the requirements of “reference standard,” but the de- same ISO quality standard, in addi- the Controlled Substance Act and gree to which that material is char- tion to ISO 17034:2016, which focuses any state enforcement agency, as acterized defines if the reference specifi cally on an evaluation of pro- well as the terms and conditions of standard is an acceptable calibra- cess uncertainty. This value is associ- any Institutional Research or Analyt- tor for the intended application. ated with manufacturing as a whole, ical Licenses or Registrations issued Simply because DEA sanctions and is reported along with orthog- in accordance with proposed labora- have been provided to a particu- onal testing results, in the reference tory activities. Sanctioned DEA-ex- lar reference standard manufactur- standard traceable documentation emptions are granted for cannabi- er, it does not automatically ensure (certifi cate of analysis [CoA]) provid- noid preparations after approval by that the reference standards pro- ed as per ISO 9001:2015. These ISO the DEA and Local State Codes to duced by that manufacturer meet accreditations offer a foundation of registered manufacturers that sup- the quality and reliability specifica- integrity when selecting a manufac- ply these solutions for laboratory or tions for laboratories testing canna- tured cannabinoid reference stand- scientifi c research purposes (2). bis samples for potency. ard material supplier.

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Only two out of the three ∆9-THC DEA-exempt ampouled formulations were within the concentration

0.018 0.018 range reported on the respective 9 0.016 CBD 0.016 Δ THC CoA when compared to the linear re- 0.014 0.014 gression of the ∆9-THC isolate. The 0.012 0.012

0.010 0.010 UV-PDA spectral purity and MS re-

0.008 0.008 9

Absorbance (AU) sponse for ∆ -THC in the formulation 0.006 0.006 and the isolate were comparable. The 0.004 0.004 reference standard formulation that 0.002 0.002

0.000 0.000 lies outside the CoA concentration 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 range does not immediately imply Time (min) Time (min) that the manufacturer provided an inaccurate reference standard concentration assessment. It is a reminder 9 Figure 1: Overlay of DEA-nonexempt CBD and ∆ -THC isolate starting materials (black) that qualifi cation of an incoming ref- with DEA-exempt ampouled reference standards from three manufacturers (green, red, blue) when separated using a validated chromatographic testing method for assay. erence standard should not be made based upon a single testing result. In this study, results were generated for Evaluation of Incoming standards using the same chroma- only one reference standard vial. It is Reference Standards tographic platform (Figure 1) (3). recommended that replicate vials are Accurate determination of chro- The CBD, ∆9-THC isolate start- analyzed from the same lot, and that matographic purity depends upon ing materials, and the ampouled ref- conclusions are made based on an the ability of an analytical method erence standards were confi rmed average concentration determination to resolve the analyte from close- to be spectrally pure by ultravio- and standard deviation value for the ly eluted impurities. Since canna- let-photodiode array (UV-PDA) de- incoming lot. bis testing methods have yet to tection, and the mass was con- Alternatively, an evaluation of ac- be standardized within the indus- fi rmed by mass spectrometry (MS) ceptability can also be made by try, the ability of chromatograph- using the in-house validated chro- comparing the average response ic methods to separate the refer- matographic method. From the lin- factor (RF) (equation 1) of the incom- ence standard peak of interest from ear response of a CBD isolate ref- ing reference standard to the RF ob- the impurities may not be consist- erence standard, the concentration served in the historical data for previ- ent between laboratories. Because of CBD in the DEA-exempt am- ous reference standards of the same of this lack of standardization, chro- pouled preparations was calculated analyte. This approach requires that matographic assay results deter- and compared to the CoA. Concen- cannabis testing laboratories com- mined for the reference standard trations of CBD were 1.013 mg/mL, pile average RF and concentration solution in a sample testing labora- 1.001 mg/mL, and 1.001 mg/mL. All information over time for incoming tory may be different than the chro- values were within the concentra- lots, and defi ne acceptable incoming matographic assay value (mg/mL) tion range specifi ed by the manufac- RF tolerance limits. With this tech- reported in the CoA. turer CoA at 1.002 ± 0.011 mg/mL, nique, it is important to use the same To demonstrate, three certified 1.002 ± 0.006 mg/mL, and 1.000 ± chromatographic separation meth- DEA-exempt ampouled canna- 0.005 mg/mL, respectively. From od, confi rmed to be consistent and bidiol (CBD) and ∆9-THC reference the concentration values, it was con- robust over time by fulfi lling method standards formulated in methanol cluded that although the references system suitability criteria (that is, re- (1.0 mg/mL) were purchased from standard solutions contained differ- covery and resolution). different certified manufacturers ent impurities, the in-house validat- Equation 1 shows the average re- (sources withheld). The chromato- ed method provided a comparable sponse factor (RF): graphic profiles of the CBD and ∆9- chromatographic separation of CBD THC DEA-nonexempt isolate start- from the impurities when compared Peak area (μV * s) RF = ing materials were compared to to the chromatographic assay meth- mg Concentration ( ) DEA-exempt ampouled reference od used to prepare the CoA. mL [1]

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regulate the volume measurement. Upward movement of the piston pro- (a) (b) duces partial vacuum in the tip, which 3.5E+06 4.0E+06 causes a metered amount of the sam- y = 3e6x + 5.44e5 y = 3e6x + 1.46e4 3.5E+06 3.0E+06 R2 = 0.8763 R2 = 0.9999 ple to be aspirated into the tip. The

3.0E+06 3.5E+06 air-cushion acts like an elastic spring

2.5E+06 2.0E+06 from which the volume of liquid in 2.0E+06 1.5E+06 the tip is suspended (Figure 3a). The 1.5E+06

1.0E+06 volume of the air-cushion, which var- 1.0E+06 ies depending on the density, va- 5.0E+05 5.0E+05 por pressure, viscosity, and temper- 0.0E+00 0.0E+00 0 0.2 0.4 0.6 0.8 1 1.2 0 0.2 0.4 0.6 0.8 1 1.2 ature of the liquid being pipetted (4,5), is most often calibrated with distilled water as the test medium. Sol- vents with vapor pressure different Figure 2: DEA-exempt CBD reference material standard curves generated using from that of water, such as those uti- (a) air-displacement and (b) glass volumetric or positive-displacement pipettes. Replicate curves were prepared by multiple analysts to confi rm linear regression trends. lized for commonly used cannabinoid solvents, push against the calibrated air-cushion forcing it to expand. Preparation of Reference Figure 2a exhibits the standard The aspirated solution is then dis- Standards for Sample Testing curve generated for DEA-exempt placed from the pipette tip via drip- In the testing laboratory, reference CBD reference standard using the ping. This phenomenon compromis- standards and sample solutions often air interface pipette. Although sever- es the accuracy and precision of the require dilution of the initial stock so- al of the data points in the serial di- dispensed volume (5). Additionally, lution to prepare a working standard. lution sequence appear to follow a commonly used solvents for cannab- When working with solvent-based so- linear trend, the curve was not linear inoids such as methanol, ethanol, or lutions such as cannabinoid formu- by statistical regression analysis (R2 = hexane have a lower density than that lations, it is important to select a pi- 0.8763). There is no justifi cation to ex- of water. Thus, the volume of liquid pette that will maintain accuracy. In clude the data points that do not sta- pipetted with an air-displacement pi- this study, cannabinoid DEA-exempt tistically conform to the linear regres- pette is systematically high when us- and isolate reference standards were sion line (y = mx + b). The results were ing these solvents neat, or as a part prepared via serial dilution using vol- replicated by two analysts with two of a mixture, if the air-cushion pipette umetric pipetting devices common different air-displacement pipettes to was calibrated with water. in testing laboratories. Calibration confi rm the nonlinear trend was con- The physical infl uences that im- curves for CBD and ∆9-THC were pre- sistent for this type of pipette. pact an air-cushion pipette are not pared using a glass volumetric pi- In contrast, for calibration curves applicable to positive-displacement pette, an air interface pipette, and prepared with a positive-displace- or glass pipettes. These pipettes re- a positive-displacement syringe pi- ment syringe pipette, or a glass vol- peatedly generated serial dilutions pette using a 1.0-mg/mL cannabi- umetric pipette, the data points that were statistically linear. Positive- noid reference standard formulation. were repeatedly statistically linear displacement pipettes operate using Practical precautions were used to (R2 ≥ 0.999), as demonstrated for CBD an integrated piston, or internal sy- reduce error, such as using calibrat- in Figure 2b. ringe. The pipette tip is immersed in ed pipettes, prewetting the pipette Differences in linearity between the liquid and the piston rod moves tip with sample solution, working at the pipetting methods can be under- down to displace the volume of air a constant temperature, pipetting in stood by considering the mechanism corresponding to the desired vol- a fully upright position, and avoiding of operation and how it is impacted ume of the pipetted liquid. As the transfer of the solution on the outside by the physical properties (that is, piston is released, the liquid is drawn of the pipette tip to the collection density and vapor pressure) of the so- into the tip. The piston comes into vessel. The standard curves for linear lution. For example, air-displacement direct contact with the liquid, leav- regression were prepared to show the pipettes rely upon the movement of ing no air-space (Figure 3b). This pi- accuracy of each pipetting method. a piston and calibrated air-cushion to petting method is suitable for both

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solvent-based cannabinoid reference standards and highly viscous liquids, Internal such as extracted cannabis oils. (a) piston (b) When using glass volumetric pipettes, although the pipetted solution comes in contact with the air- space, the meniscus is visually titrated Plunger against a volumetric calibration mark. Delivery time and waiting time de- Sample termine pipetting accuracy. The de- Sample livery time is the period of time after the suction is released for the meniscus to fall from the upper volumetric Disposable Air-space piston mark to the pipette tip. Waiting time begins after the liquid reaches the tip and comes to rest (6). Both must Figure 3: Comparison of pipetting mechanisms for (a) air-displacement and (b) positive- displacement pipettes. be adequately used to ensure accurate pipetting, no matter the physical properties of the liquid. 1.85 Self-sealing septum Storage and 1.75 In-Use Considerations 1.65 Instrumentation and consumables are 1.55 the largest expense when operating a 1.45 testing laboratory. Consumables are Punctured 1.35 septum Self-sealing after puncture

defi ned as items that are used for an in- Concentration 1.25 tended purpose and must be replaced Degraded septum after because they wear out or are used up 1.15 puncture (7). They include both reference stand- 1.05 ards and sample testing vials (8). Can- 0.95 0510 15 20 25 30 nabinoid reference standards are pack- 35 Time after injection (h) aged in glass ampoules sealed under inert gas to promote stability by pre- Figure 4: Plot of solution concentration for CBD reference standard stored in a vial with venting evaporation and degradation a degraded septum compared with a PTFE–silicon self-sealing septum. from air and light during transport (9). After the ampoule has been opened in vials may be more economical, sur- the manufacturing process. For exam- the laboratory, a key concern is the po- face-treated inert glass vials can be ple, cracks in the cap may result after tential for degradation during gener- certifi ed as chemically clean by the stretching over an opening that is too al use. A decrease in purity because of manufacturer. This certifi cation min- large, or for a smaller opening, the cap chemical degradation is always a con- imizes the potential for chromato- may not provide an adequate air seal. cern, but for purposes of this manu- graphic anomalies caused by manu- Both result in the escape of the solvent script, the solutions were stored before facturing process contamination, and from the reference standard solution, use at the temperature recommended is especially important for samples in- resulting in an increase in the cannab- in the package insert. tended for low-level impurity analysis inoid concentration. Although it may It is common practice to transfer by mass spectrometry. take a few extra seconds to secure a reference standard solutions from The most common closures for cap with screw threads, the threads the shipping ampoules to liquid chro- these LC vials are caps that snap or provide adequate mechanical force to matography (LC) vials for use and screw onto the vial. Snap caps provide hold the septum in place and provide storage. LC vials are typically man- an advantage because they are easy a consistent air seal. ufactured from polypropylene plas- and fast to assemble. Glass LC vials There is an overwhelming varie- tic or glass. Although polypropylene may show imperfections because of ty of cap septum offerings including

VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 www.CannabisScienceTech.com | CANNABIS SCIENCE AND TECHNOLOGY 28 Feature

rubber, polyethylene, polypropylene, in the sample manager at 20 °C. References silicone, and polytetrafl uoroethylene The DEA-exempt reference stand- 1) United States Department of Justice, (PTFE). The challenge is determining ard CBD concentration inside the vial Drug Enforcement Administration, which is appropriate for solvent-based that used the self-sealing PTFE–sili- Diversion Control Division. cannabinoid solutions. Rubber and sil- cone septum remained stable after Controlled Substance Schedule: Marijuana. https://www.deadiversion. icone alone are not recommended be- 1.5 days of replicate injections (Fig- usdoj.gov/schedules/#defi ne. cause they may express extractables ure 4). A hole was observed in the less (accessed Oct 17, 2017). and leachables when in contact with expensive, solvent-incompatible sep- 2) United States Department of Justice, solvents. Chemically resistant polyeth- tum after collecting data at two injec- Drug Enforcement Administration ylene and polypropylene septa are in- tion time points. The coring resulted in 21 CFR Part 1301 [Docket No. tended for single injection, aqueous- evaporation of the reference standard DEA–447] “Applications to Become Registered Under the Controlled based sample mixtures. Although solvent from the cannabinoid over 1.5 Substances Act To Manufacture polypropylene septa offer improved days, resulting in concentration of the Marijuana To Supply Researchers solvent compatibility compared to solution. It is anticipated that a sample in the United States,” Federal polyethylene, the piercing force is manager without temperature control Register, 81(156), August 12, 2016. slightly higher, resulting in an increase capabilities would likely result in an 3) C. Layton and A.J. Aubin, in the probability of coring. Pure PTFE even greater increase in solvent evap- J. Liq. Chromatogr. Relat. septa are not resealable and should oration through the open septum. Technol. 41, 114–121 (2018). not be used with highly volatile sol- It is not suggested that reference 4) K. Ewald, “Infl uence of Physical vents, short cycle times, or multiple standard solutions are stored for long Parameters on the Dispensed Volume of Air-Cushioned injection methods, all of which are periods in LC vials with septum. They Pipette,” Eppendorf User common when testing cannabinoids. can be stored for longer periods in Guide No. 21. June 2015. Laminated hybrid PTFE–silicone septa an LC vial with a chemically compati- 5) K. Ewald, “Fundamentals of provide the ideal solution. A thin fi lm ble septumless storage cap to reduce Dispensing,” Eppendorf User of PTFE laminated to the side of the solvent loss (11). Even with this precau- Guide No. 19. June 2015. septum that faces the sample limits tion, one must consider the number of 6) “Volumetric Measurement in the exposure of the silicone elastomer to times the vial is opened and closed to Laboratory,” www.Brand.com. the cannabinoid solvent, while the sil- minimize solvent evaporation. As with (accessed January 16, 2018). icone on the outside surface provides incoming reference standards, the RF 7) “Consumables,” www.Investopedia. resealing capability (10). Additional- value can be monitored to determine com. (accessed June 6, 2018). ly, these hybrid septa are offered in a if the standard is fi t for use after stor- 8) J. Witonsky, “Laboratory Spending preslit format to eliminate vacuum for- age based on its concentration. Trends,” Lab Manager (2011). mation or coring as it is pierced by the 9) M. Rettinger, “Potency Claims – injection needle. Conclusions Only as Accurate as Your Reference To demonstrate how quickly the ref- Cannabis testing laboratories rely di- Material,” Cerilliant Corporation, erence standard solution can be af- rectly on solvent based DEA-exempt Emerald Conference, 2016. fected by an inadequately sealed vial, a cannabinoid reference standards to 10) Chromatography Columns DEA-exempt CBD reference standard accurately quantify cannabis samples. and Consumables, “Septum was removed from the sealed ampoule After selecting a reputable supplier, Selection Guide,” Thermo Scientifi c, 2014–2015. and aliquoted into two, certifi ed, Ac- laboratories should consider adopt- quity 12 x 32 mm sample manager LC ing a qualifi cation process for the in- 11) “Waters Quality Parts, vials (Waters Corporation). Both vials coming reference standards to ensure Chromatography Columns and Supplies Catalog,” Waters were sealed with screw-cap closures. incoming formulations meet in-house Corporation, 2015–2016. One screw cap contained an inex- assay specifi cations. Because cannab- pensive solvent-incompatible septum inoid reference standard solutions are susceptible to coring (source with- solvent based, laboratories must be held), and the other contained a pres- diligent when handling these formula- Catharine E. Layton and Andrew lit PFTE–silicone self-sealing septum tions (that is, pipetting, vial storage) to J. Aubin are with Waters Corpo- (Waters). Replicate injections were per- promote integrity of the concentration ration in Milford, Massachusetts. Direct correspondence to: Catharine_ formed from each vial over the course from which potency determinations [email protected] of 1.5 days while the vials were stored are established.

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30 Feature

Leveraging Selectivity and Effi ciency to Take the Strain Out of LC–UV Method Development for Cannabinoid Profi ling

More than 100 cannabinoids have been isolated from cannabis in addition to the fi ve most commonly tested: ∆9-tetrahydrocannabinol (∆9-THC), ∆9-tetrahydrocannabinolic acid (THCA), cannabidiol (CBD), cannabidiolic acid (CBDA), and cannabinol (CBN). Although many methods have been published that show the separation of these major cannabinoids, most do not take into account the possibility of interference from minor cannabinoids. This interference is most problematic in concentrates where minor cannabinoids can be enriched to detectable levels. Additionally, some terpenes absorb ultraviolet (UV) light at the same wavelength as cannabinoids, which can result in an additional source of interference. In this study, the liquid chromatography (LC)–UV separation of 16 cannabinoids of interest was performed while the potential impact from minor cannabinoids and terpenes on reported potency values was monitored. The method was applied to commercially available CBD oils that have recently become suspect because of inaccurate label claims. Justin Steimling

ust as consumers rely on nutrition facts labels on (∆9-THC) have been found to be very effective for the treat- food for making informed choices about their diets, ment of various medical disorders; however, accurate dos- Jlabels on cannabis products are intended to provide ing is required for positive patient outcomes. As exam- critical information for accurate dosing. Consumer confi - ples, CBD was determined to be an effective treatment dence in labeling claims can be shattered by reports of infor those suffering from treatment-resistant epilepsy while accuracies. A recent study published by the Journal of the ∆9-THC has been found to be an effective treatment for American Medical Association showed that of 84 canna- pain when coadministered with reduced doses of opioids bidiol (CBD) products purchased online, only 31% were ac- (2,3). Whether labeling inaccuracies stem from inadequate curately labeled (1). This labeling problem is alarming since analytical methods, instability of formulations, or surrepti- 9 cannabinoids such as CBD and ∆ -tetrahydrocannabinol tious laboratory practices, all result in the same potentially TALI RUSS/SHUTTERSTOCK.COM

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tetrahydrocannabivarinic acid (THC- 1 VA), cannabinolic acid (CBNA), ∆8-tetrahydrocannabinol (∆8-THC), cannabi- cyclol (CBL), cannabichromene (CBC), 3 and cannabichromenic acid (CBCA). 4 A 150 mm x 4.6 mm C18-type col-

2 9 umn packed with 2.7-μm superfi cial-

8 ly porous particles (SPPs) was initially 5 6 14 scouted using popular mobile phas- 7 es for the analysis of cannabinoids 10 15 11 12 13 (water and acetonitrile modifi ed with 16 0.1% formic acid) with UV detection at 228 nm. C18-type columns are ide- 0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 Time (min) al for the separation of cannabinoids because of their hydrophobic and Figure 1: The separation of 16 cannabinoids by HPLC–UV. Column: 150 mm x 4.6 mm, shape-selective characteristics. The 2.7-μm Raptor ARC-18; mobile-phase A: water, 0.1% formic acid (v/v), 5 mM ammonium combination of the appropriate sta- formate; mobile-phase B: acetonitrile, 0.1% formic acid (v/v); elution: isocratic at 75% B tionary phase with SPPs increases the over 9 min; fl ow rate = 1.5 mL/min; injection volume: 5 μL; oven temperature: 30 °C; detection: UV absorbance at 228 nm. Peaks: 1 = CBDVA, 2 = CBDV, 3 = CBDA, 4 = CBGA, speed and improves resolution com- 5 = CBG, 6 = CBD, 7 = THCV, 8 = THCVA, 9 = CBN, 10 = CBNA, 11 = ∆9-THC, 12 = pared to traditional fully porous par- ∆8-THC, 13 = CBL, 14 = CBC, 15 = ∆9-THCA, 16 = CBCA. ticles (FPPs) of the same particle size. The use of a 150 mm x 4.6 mm ana- negative impact to the health and including terpenes, fatty acids, sug- lytical column allows for the most ef- safety of consumers. ars, fl avonoids, and pigments (4). The fi cient separations under the con- Although the state of analytical introduction of cannabis concentrates straints of legacy instrumentation. testing in the cannabis industry has into additional matrices to create can- Using isocratic conditions it was improved since the inception of le- nabis-infused products further com- found that 14 of the 16 cannabinoids galized medical cannabis, there are plicates analysis. If product labels are could be baseline resolved; however, still many opportunities to update ex- to accurately refl ect cannabinoid con- CBNA was coeluted with ∆9-THC re- isting methods in this dynamic mar- tent, the complete sample composi- gardless of fl ow rate, column temper- ket. Historically, interest in cannabi- tion must be considered in develop- ature, or mobile-phase composition. noids typically focused on THC, CBD, ing a robust and accurate analytical In an attempt to separate CBNA and their carboxylated forms, but the method. Herein, the process of de- from ∆9-THC, the pH of the mo- analysis of more-abundant minor can- veloping a complete workfl ow solu- bile phases was adjusted by using nabinoids is starting to gather mo- tion for the analysis of cannabinoids 0.1% acetic modifi ed mobile phases mentum. Testing laboratories most is discussed with emphasis on LC–UV in place of formic acid. This strategy frequently use liquid chromatography conditions, sample preparation strat- was attempted in an effort to take ad-

(LC)-based techniques paired with ul- egies, method applications, and high- vantage of the pKa of CBNA because traviolet (UV) detection for cannab- throughput enabling technologies. it would be less retained in its ion- inoid profi ling because of its low ini- ized form, but THC would not be im- tial cost, ease of use, and robustness. Development of LC pacted as a neutral compound. Ion- Because UV detection is limited in its Method Conditions ized compounds are more polar and spectral deconvolution abilities, chro- Method development began with not as strongly retained by reversed- matography plays a critical role in not the goal of achieving baseline resolu- phase mechanisms. The use of acetic only cannabinoid identifi cation, but tion for the following 16 cannabinoids acid maintained the same selectivity also accurate quantitation. The de- by LC–UV: ∆9-THC, ∆9-tetrahydro- and resolution of the original separa- mands on the analytical column are cannabinolic acid (THCA), CBD, can- tion, but still resulted in the coelution only made more diffi cult considering nabidiolic acid (CBDA), cannabinol of CBNA and ∆9-THC. The pH of the the complexity of the matrix. Canna- (CBN), cannabidivarinic acid (CBDVA), mobile-phase system was further in- bis is a complex matrix that contains cannabidivarin (CBDV), cannabigerol- creased by the use of water modifi ed more than 100 cannabinoids as well as ic acid (CBGA), cannabigerol (CBG), with 5 mM ammonium formate and un- hundreds of additional components tetrahydrocannabivarin (THCV), modifi ed acetonitrile. This approach

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Table I: Inter-run accuracy and precision (n = 6)

QC LLOQ QC Low QC Mid QC High QC Dilution (20-fold)

5.00 μg/mL 30.0 μg/mL 150 μg/mL 400 μg/mL 1000 μg/mL

Analyte Avg. Avg. Avg. Avg. Avg. Avg. Avg. Avg. Avg. Avg. Conc. Accu- % Conc. Accu- % Conc. Accu- % Conc. Accu- % Conc. Accu- % (μg/ racy RSD (μg/ racy RSD (μg/ racy RSD (μg/ racy RSD (μg/ racy RSD mL) (%) mL) (%) mL) (%) mL) (%) mL) (%) CBD 4.96 99.2 3.14 30.8 103 1.84 156 104 0.455 396 99.0 0.455 1,036 104 0.341 resulted in a loss of retention for all of CBNA from ∆9-THC while still main- Additional Sources acidic cannabinoids because of the taining the separation of the remain- of Interference ionization of carboxylic acid func- ing 14 cannabinoids. To this point, a During LC method development, it tional groups. To reach an interme- column oven temperature of 40 °C was determined that minor cannab- diate pH, the aqueous mobile phase had been used for method develop- inoids can affect the quantitation of was modifi ed with both 0.1% formic ment. To enhance the shape selective the major cannabinoids CBD and ∆9- acid and 5 mM ammonium formate, characteristics of the stationary phase, THC. Another class of compounds whereas the organic mobile phase was the column oven temperature was re- found in cannabis, terpenes, can also modifi ed with only 0.1% formic acid. duced to 30 °C. This adjustment re- be present in high enough concentra- These mobile phases combined with sulted in the baseline separation of all tions to interfere with sample analy- a 1.5 mL/min fl ow rate using 75% or- 16 cannabinoids with a complete cycle sis. A mix of 21 common terpenes ganic allowed for the partial resolution time of 9 min (Figure 1). found in cannabis was prepared and

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Table II: Determined concentrations of CBD in commercially available hemp-derived CBD oils

CBD, Label ±10% Range CBD, Actual Product Name Product Type Claim (mg/mL) (mg/mL) Concentration (mg/mL) Product 1 Vape oil 3.33 3.00–3.66 2.05 Product 2 Vape oil 6.67 6.00–7.34 5.59 Product 3 Vape oil 500 450–550 0.242 Product 4 Sublingual oil 11.0 (hemp extract) NA 6.24 Product 5 Sublingual oil 6.11 5.50–6.72 5.21 Product 6 Raw hemp oil 30.0 27.0–33.0 18.4

extracted using 950 μL of metha- 1 nol followed by vortexing for 30 s at 3000 rpm. After that, 750 μL of the sample was then mixed with 250 μL of 3 4 water followed by vortexing for 10 s at 3000 rpm. Finally, 400 μL of the sample 9 2 was fi ltered using a Thomson Single 8 Step standard fi lter vial with a 0.2-μm 14 56 polyvinylidene fl uoride (PVDF) mem- 7 brane before LC–UV analysis. 10 15 11 12 13 16 Results and Discussion Using linear 1/x weighted regression, the method showed good line- 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.003.20 3.40 3.60 3.80 4.00 2 Time (min) arity for CBD with an r value of 0.999. The method accuracy was demon- Figure 2: The separation of 16 cannabinoids by UHPLC–UV. Column: 100 mm x 3.0 mm, 1.8-μm strated to be within 4% of the nominal Raptor ARC-18; mobile-phase A: water, 0.1% formic acid (v/v), 5 mM ammonium formate; mobile- concentration for all QC levels. The phase B: acetonitrile, 0.1% formic acid (v/v); elution: isocratic at 75% B over 4 min; fl ow rate = 1.0 mL/min; injection volume: 1 μL; oven temperature: 30 °C; detection: UV absorbance at 228 nm. percent relative standard deviation Peaks: 1 = CBDVA, 2 = CBDV, 3 = CBDA, 4 = CBGA, 5 = CBG, 6 = CBD, 7 = THCV, 8 = THCVA, (%RSD) was within 3.14% for all QC 9 = CBN, 10 = CBNA, 11 = ∆9-THC, 12 = ∆8-THC, 13 = CBL, 14 = CBC, 15 = THCA, 16 = CBCA. levels indicating good method precision (Table I). According to California injected using the same conditions Sample Preparation regulations, a cannabis product must as previously developed for cannab- The utility of the chromatographic not differ from the labeled concen- inoids. Fortunately, the absorbance method was evaluated by perform- tration of CBD by ±10% (5). Applying profi le for most terpenes differs from ing quantitative analysis of CBD in this labeling guideline to the samples cannabinoids such that they do not commercially available hemp-derived evaluated in this study results in the result in over reporting for cannabi- CBD oils. Calibration standards and possibility of all six products not be- noids. The terpenes that absorb UV quality control (QC) samples were ing in compliance with these regula- light best at 228 nm, ocimene and prepared in 25:75 water–methanol tions (Table II). One product failure is its isomers, do not interfere with the across a linear range of 5–500 μg/mL. attributed strictly to labeling nomen- quantitation of any monitored can- A QC dilution sample was prepared clature as the CBD content is not di- nabinoid. Minor terpene interfer- at 1000 μg/mL and diluted 20-fold rectly stated. Vape oils did not con- ences were found that could impact to demonstrate the validity of dilut- tain a signifi cant amount of CBDA to the quantitation of CBGA and THC- ing samples into the linear range. account for the discrepancy in the la- VA if present in substantial concen- Six hemp-derived CBD oils were ob- beling. The large discrepancy in the trations, but the quantitative analysis tained for analysis. These products result for Product 3 is most likely be- of these cannabinoids is not current- consisted of three vape oils, two cause of photodegradation from im- ly required in states such as California sublingual oils, and one raw hemp proper packaging. In addition to and Colorado (5,6). oil. Next, 50 μL of CBD product was CBD, other cannabinoids were found

CANNABIS SCIENCE AND TECHNOLOGY | www.CannabisScienceTech.com VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 Featture 35

in most samples. Additional cannabi- column packed with 1.8-μm SPPs of CBD oils. As interest in expanded can- noids that were found included: CB- the same stationary phase, the sepa- nabinoid profi les continues to grow, DVA, CBDV, CBDA, CBG, ∆9-THC, ration can be achieved in 4 min (Fig- selectivity and effi ciency can continue ∆8-THC, CBN, CBNA, and CBC. The ure 2). This approach results in a cy- to be leveraged to remove the strain concentrations of these cannabinoids cle time that is more than twice as fast of LC–UV method development. were not quantitatively evaluated be- with a 70% reduction in consumed cause of their relative low abundance, mobile phase per sample. Columns References 9 but the concentration of ∆ -THC in all must be effectively paired to an ap- 1) M.O. Bonn-Miller, M.J.E. Lofl in, B.F. samples appeared to be well below propriate LC system that is not only Thomas, J.P. Marcu, T. Hyke, and 0.3% w/w based upon the peak height robust at high system back pressures, R. Vandrey, JAMA, J. Am. Med. of a 50-μg/mL standard allowing the but also optimized to reduce extra Assoc. 318, 1708–1709 (2017). use of THC-free labels. column volumes. Any dispersion that 2) O. Devinsky, E. Marsh, D. Friedman, To ensure that sample results were occurs in the valves, connecting tub- E. Thiele, L. Laux, J. Sullivan, I. Miller, R. Flamini, A. Wilfong, F. not skewed because of poor extrac- ing, and fl ow cell will reduce the ef- Filloux, M. Wong, N. Tilton, P. tion effi ciencies, a recovery exper- fi ciency and the resolution of sepa- Bruno, J. Bluvstein, J. Hedlund, R. iment was performed by spiking rations. It is particularly important to Kamens, J. Maclean, S. Nangia, each product type with an addition- use low-volume fl ow cells (approxi- N. Shah Singhal, C.A. Wilson, A. Patel, and M. Roberta Cilio, Lancet al 2.00 mg/mL of CBD. Recovery re- mately 1 μL) because of the relative- Neurol. 15, 270–278 (2016). sults for vape oil, sublingual oil, and ly large volumes present in standard raw hemp oil were 102%, 98.5%, and fl ow cells (8). 3) S. Nielsen, P. Sabioni, J.M. Trigo, M.A. Ware, B.D. Betz-Stablein, B. 105%, respectively. Murnion, N. Lintzeris, K. Eng Khor, With regard to sample preparation, Conclusions M. Farrell, A. Smith, and B. Le it should be emphasized that the ad- Considering the structural similarities Foll, Neuropyschopharmacology dition of water following extraction of cannabinoids, it can be diffi cult to 42, 1752–1765 (2017). with methanol is critical to precipitate develop a robust analytical method 4) M. ElSohly, W. Gul, in Handbook of lipids that are present in raw hemp oils that is capable of producing accurate Cannabis,1st Ed., R.J. Pertwee, Ed. (Oxford University Press, Oxford, and common carrier oils for sublin- results. Increasing interest in the anal- United Kingdom, 2014), pp. 3–22. gual products (that is, coconut medi- ysis of minor cannabinoids can further um-chain triglycerides [MCT] oil, sun- complicate the separation. Fortunate- 5) California Code of Regulations, Title 16, Division 42, “Chapter fl ower oil, sesame oil, and so forth). ly, demanding separations can still be 6, Section 5724 – Cannabinoid Failure to perform this step will cause achieved through deliberate method Testing,” (Adopted May 5, 2018; sample precipitation to occur once in- development by leveraging selectivity Effective June 6, 2018), p. 105. troduced into the mobile phase dur- and effi ciency. The power of selectiv- 6) Code of Colorado Regulations, ing high performance liquid chroma- ity is realized through changes to the 1 CCR 212-1, “Section M 1503 – tography (HPLC) analysis. The inlet frit mobile phase that can alter the pH Medical Marijuana Testing Program – Potency Testing,” (Adopted of the analytical column will quickly enough for a critical separation to oc- February 21, 2018; Effective clog resulting in elevated back pres- cur or a change in oven temperature February 21, 2018), pp. 218–221. sures and decreased column lifetime. that can change the characteristics 7) E.M. Mudge, S.J. Murch, and It is important to not combine the ad- of a stationary phase. The increased P.N. Brown, Anal. Bioanal. Chem. dition of water into the extraction step effi ciencies of SPPs compared to ful- 409, 3153–3163 (2017). with methanol because this has been ly porous particles of the same par- 8) J. De Vos, K. Broeckhoven, shown to result in reduced recoveries ticle size enables faster separations. and S. Eeltink, Anal. Chem. for cannabinoids in cannabis oils (7). Pairing sub-2-μm SPPs with appropri- 88, 262–278 (2016). ate instrumentation is critical for those Ultrahigh-Pressure Liquid laboratories interested in pushing the Chromatography Analysis limits of sample throughput. It was To improve laboratory throughput, demonstrated that 16 cannabinoids Justin Steimling the effi ciency of sub-2 μm SPPs can be of interest can be separated in 9 min is an Applications Manager, LC Solu- used to increase the speed of analy- on a traditional HPLC system. The de- tions at Restek Corporation in Belle- sis while maintaining the same reso- veloped method was compatible for fonte, Pennsylvania. Direct correspondence to: lution. By pairing an appropriate sys- the identifi cation of cannabinoids and [email protected] tem to a 100 mm x 3.0 mm analytical quantitation of CBD in hemp-derived

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Certifi ed Reference Material Manufacturing Challenges

Some analysts will look at a list of pesticides or solvents and visualize adding small amounts of pure material of each analyte into a volumetric fl ask and then bringing the fl ask up to volume with their solvent of choice. In rare instances this technique will work, but not often. Prior to making a mixture, factors such as solubility and reactivity must be considered. Are all of the analytes soluble in the solvent that I want to use? Do I need to use an intermediate solvent? Will the analytes react with each other or will they react with the solvents? Do I need to add a preservative? Which preservative should I use and will any of the analytes react negatively to the preservative? The answers to these questions come from experience and a working knowledge of the chemistry of reference material (RM) production. Manufacturing the standard mixture is only the fi rst step in creating a certifi ed reference material (CRM). The CRM must be characterized using a metrologically valid procedure. A certifi cate of analysis is produced listing certifi ed values for specifi ed properties and all analytes, including a calculation of total uncertainty, homogeneity, stability under specifi ed conditions, and metrological traceability. The certifi cate of analysis is delivered with the standard.

Don Shelly

f you work in an analytical chemistry laboratory you CRM Challenges may have heard the terms certifi ed reference materi- All CRM manufacturers strive for the lowest possible com- Ial (CRM) and reference material (RM). An RM typically bined uncertainty during the manufacturing process. There comes with a certifi cate of composition (COC) that states are some variables that cannot be precisely controlled dur- the identity of the product, the purity of the product, and ing the manufacturing process such as the limitations of the details of its characterization. In addition to the infor- Class A glassware (typically, the uncertainty at a specifi c mation found in the RM COC, the CRM certifi cate of anal- temperature is written on the glassware) and syringes, pre- ysis (COA) declares that the material characteristics were cise room temperature, and the uncertainty inherent in all determined by a metrologically valid procedure for the analytical balances despite calibration with National Insti- specifi ed properties and contains a statement of metro- tute of Standards and Technology (NIST) certifi ed weights. logical traceability, homogeneity data, long term stabil- The most challenging uncertainty to calculate is the uncer- ity results, and all factors that determine the CRMs total tainty obtained from possible analyte to analyte and analyte combined uncertainty, to name a few. This uncertainty in- to solvent interactions. cludes the measured uncertainty inherently found in all measurement tools used in the manufacturing process What Skills Are Needed to Create CRMs? as well as the effects of storage, transportation condi- The fi rst skill required when making a stock standard from tions, and possible chemical interactions affecting long a neat material is knowing which solvent is miscible or solu- term stability. ble with the analyte and if the analyte and solvent will quickly interact under the given storage conditions. If your work- The Benefi ts of CRMs ing standard will be in a solvent different from your stock CRMs, as opposed to RMs, give labs a competitive advan- solvent, then the stock’s solvent is sometimes known as an tage and peace-of-mind. ISO 17025 recommends that ac- intermediate. For example, if you want to spike used motor credited labs use CRMs whenever available. As a laborato- oil into water, you know that oil and water aren’t miscible. If ry, you can trust that everything has been done to provide you fi rst dilute your oil with isopropanol at a reasonable con- you with a high quality standard that helps to ensure that centration, the oil–isopropanol solution can be successfully your data can be generated correctly the fi rst time, and spiked into the water matrix and you will have the used mo- every time. After all, the quality of your data can direct- tor oil in solution with your water. The isopropanol is your in- ly affect consumer safety. Because the use of CRMs helps termediate solvent. to ensure data quality, your services are of greater value to You have created all of your individual stock standards and your clients and prospective clients. now you want to combine portions of each stock to make

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your working standard mixture. Again, the number and concentration of your (rate of degradation) increases as envi- you need to ask, are the solvents mis- analytes in a mixture. Many laboratories ronmental conditions become harsher cible and will my analytes stay in so- request CRMs with all possible analytes compared to the stated storage condi- lution? Will that solution be homoge- at a high concentration in one ampule tions. This ensures that the certifi ed val- nous? Will the analytes quickly interact for cost savings. When your CRM ex- ues for the analytes are within the stat- with each other or with the solvents? pires in a couple of months, are you sav- ed uncertainties for the specifi ed shelf To answer these questions you need ing money? More organic compounds life. The method is typically performed experience. If you have done this be- at higher concentrations mean there by heat stressing the standard. Several fore you will know what is going to hap- are greater chances for fast chemical in- ampules of the standard are made and pen. If this is your fi rst time making this teractions to occur. Use a stock product one is placed in a storage unit at the cor- mixture, you won’t know for sure un- or, if a custom one is required, keep the rect temperature. The other ampules til you have fi nished the mixture and concentrations reasonable and allow are placed in heated environments at some time has passed. I will give you the manufacturer the option of separat- three or more temperatures between a hint: All organic chemicals interact ing analytes into multiple mixtures. The the stated storage temperature and as do many metals. To minimize your analytes’ shelf life will be longer and 100 °C for a set time. The ampulized interaction and increase the shelf life, your headaches fewer. standards are analyzed sequentially us- you need to know the correct storage ing a metrologically validated method. conditions (colder is not always bet- How to Ensure Stability If recovery of any of the analytes dem- ter) and which preservative, and how Precision and accuracy are not the only onstrates a recovery of 95% or less, the much of it, to add to your mixture. Stor- concerns you might have about your standard is considered failed. This data ing a standard at too low of a temper- standards; stability is also a major factor. is entered into an Arrhenius plot to pro- ature can negatively affect homogene- The accidental use of a standard that vide a conservative estimate of shelf life. ity and possibly cause some analytes has degraded will result in costly reanal- As the product ages, retained samples to precipitate out of solution. It’s usu- ysis and increased turn-around time, se- can be pulled from storage and test- ally a good idea to sonicate or shake verely hurting your bottom line and your ed using the classical method to deter- mixtures at room temperature before laboratory’s credibility. CRM manufac- mine if the shelf life can be lengthened using, especially if they are cloudy or turers follow a set of ISO 17034 protocols or if it needs to be shortened. if you see solids fl oating in the solvent. to estimate a products shelf life. Many of you are probably now ask- There are three types of data that Summary ing yourselves, what is a preservative? can be used to determine a products As you can see, a great deal of effort is Analyte–solvent interaction can impact shelf life: historical data, classical meth- put into determining the values print- the pH of your solution and redox po- od data, and the accelerated method ed on your COAs. Even so, there can tential over time. Acetonitrile can break data. Historical data rely on the man- be unknowable aspects in the science down into acetamide as the solvent ufacturer’s knowledge and data of the of chemistry and the unforeseen can ages, which makes your solvent basic. exact reference material being made occur, but you can be assured that the Urea based pesticides will degrade un- to know how long that material can be best available technology was used der these conditions. A small amount expected to remain accurate under to certify your reference materials. To of dilute formic acid can be added to the assigned storage conditions. Using reduce the possibility of the unfore- your mixture to counter these reactions. the classical method, the manufacturer seen, I recommend resisting the urge Over time, halogenated hydrocarbons makes the product and tests retained to amend your custom or catalog mix- will interact with methanol to form ac- samples of the standard over time until tures unless truly necessary. Also, ids. Low pH will degrade linear ketones the product fails. As you can imagine, when in doubt, let the professionals do very quickly. A dilute ammonium hy- this method is very time consuming it. You will save yourself time, money, droxide solution will slow these reac- and I doubt that you want to wait years and headaches. tions. Now you know why you should to fi nd out when your standard will fail. keep your acid sensitive and base sen- Since most laboratories prefer custom sitive analytes in separate mixtures. standards, the only practical way to de- Don Shelly is the Food and Environ- These are just a couple of examples of termine shelf life is by using the accel- mental Product Manager, North Amer- analyte–solvent interactions. There are erated method. ica for LGC Standards in Manchester, New Hampshire. Direct correspon- hundreds of known interactions and The accelerated method makes the dence to: [email protected] many interactions are dependent on assumption that the product failure

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LabVantage Cannabis boosts laboratory testing productivity while reducing total cost and risk.

abVantage Cannabis is a purpose-built Laboratory Information Management System L(LIMS) that offers unique benefits to the cannabis testing industry. Product features are Marty Pittman configured to meet cannabis testing requirements, taking into account the varied regula- Senior Product Manager tory landscape. LabVantage Cannabis streamlines laboratory operations by offloading manual LabVantage Cannabis data entry and providing an electronic repository that can be easily queried for fast access to results. Cannabis Science and Technology (CST) recently sat down with Marty Pittman, senior product manager, to discuss how LabVantage Cannabis benefits testing laboratories and options for its deployment.

CST: Why do cannabis testing laboratories need a Laboratory Information Management System (LIMS)? Pittman: Initially, most laboratories find that the amount of data associated with their customers’ testing is minimal enough to be managed in spreadsheets and other paper formats. However, as their customers’ businesses grow, the amount of data that testing laboratories have to work with increases. A LIMS can resolve much of the extra workload, helping laboratories to focus more on testing and less on managing data.

CST: What is unique about the LabVantage Cannabis product, which is LabVantage Solutions’ LIMS for this industry? Pittman: LabVantage Cannabis, which is derived from our core LabVantage product, is configured for the cannabis industry. For example, American Herbal Pharmacopoeia methods that are specific to cannabis testing are included. This saves customers a lot of time in setting up and configuring the LIMS before they can make a product. LabVantage also provides a portal that cannabis growers can use to request testing directly. This feature facilitates sample collection, testing, and return of results. In addition, LabVantage provides an interface with government systems, keeping regulatory monitoring of the cannabis product and testing in one place.

CST: How do speed and cost of implementation of LabVantage Cannabis compare with that of a traditional LIMS deployment? Pittman: When implementing a LIMS out of the box, a laboratory must make many determinations. These may include deciding upon what master data must be collected to comply with regulatory requirements, process flows in the laboratory, what data must be gathered from which instruments, and the data approval process. LabVantage Solutions worked with its customers in the cannabis industry to identify these items upfront. As a result, LabVantage

SPONSORED BY CHOOSING A LABORATORY INFORMATION MANAGEMENT SYSTEM (LIMS) FOR THE CANNABIS INDUSTRY

Cannabis incorporates much of the information that would oth- customer-owned cloud environments. The customer needs erwise have to be discovered, documented, and configured an annual subscription for maintenance, but owns the product. from scratch. This allows the LabVantage Cannabis customer As such, they are responsible for providing all support and to jump right in and begin using the LIMS. testing of the operating system, application server, and database server, as well as for working with LabVantage to support CST: What features make LabVantage Cannabis unique the application itself. to cannabis testing? The SaaS model is a “pay-as-you-go” solution in which the Pittman: In the cannabis industry, there are some unique customer pays an annual subscription fee per user. In this case, requirements when requesting samples. The request does LabVantage manages the operating system, application server, not come from an internal laboratory source; external growers and database server; the customer has to deal only with the or other cannabis product manufacturers must be able to LIMS itself. This allows the customer to focus on its laboratory request the test. LabVantage Cannabis provides a portal that processes (i.e., collecting and managing data) without having allows the grower to submit the request for testing, which to worry about the behind-the-scenes hardware and operating initiates the sample collection process. LabVantage also systems needed to support the environment. includes a chain of custody that is in place when the sample Some LabVantage customers have reported that the is received so that it is processed in a known workflow. Data lower yearly cost of the SaaS model is an advantage. Other are peer reviewed to ensure that there are no transcription customers prefer the enterprise model because they can pur- errors. Finally, a Certificate of Analysis is generated. chase it upfront and then just pay for the annual subscription During the process (from sample collection to completion for maintenance. Offering these two models gives customers of the Certificate of Analysis), data must be provided to state a lot of flexibility in terms of deciding their payment schedule. governments at certain points to track the cannabis material and testing results. As I mentioned, LabVantage provides the CST: How does all of this benefit users? interface to do that. Much of the functionality that is needed for Pittman: The biggest benefit is that users can focus more on American Herbal Pharmacopoeia testing is provided upfront. their specialty—namely, the testing of laboratory materials. By In some cases, this is extended to allow for automated instru- having the LIMS in place, users worry less about transcription ment integration. This enables an instrument to produce an errors in Excel spreadsheets and where papers are located output file, which can be parsed and imported into the LIMS. in the laboratory. LabVantage gives laboratories the ability to This spares an analyst from having to manually key data from store all of their data in a central location where it can be easily the instruments into the LIMS for tracking and generation of accessed when customers have questions about testing the Certificate of Analysis. results or the status of samples. LabVantage offloads many of the day-to-day physical transactions of entering data or CST: How does LabVantage Cannabis address the moving papers from desk to desk. Users can concentrate on varied regulatory landscape? the analysis of materials and tracking of results. Pittman: The highly flexible configuration options of our core LabVantage LIMS are retained in the LabVantage Cannabis CST: Lastly, when a customer invests in LabVantage product. Customers can modify LabVantage Cannabis as Cannabis, how do they know if they are getting their needed to meet their own requirements. For example, it could money’s worth? be configured on the basis of where the customer is located, Pittman: LabVantage Cannabis confers advantages in multiple what regulatory bodies they must comply with, or what labora- areas. Customers quickly find that they can process more tory methods they have in place. samples with the same number of resources. It allows chemists to spend more time on bench-top analysis and less on CST: Please describe the two options offered for the data entry. In addition, electronic data can be mined, queried, deployment of LabVantage Cannabis. and analyzed. LabVantage Cannabis customers can drill Pittman: LabVantage has a software as a service (SaaS)- down to look for trends in their data. The ability to perform based option and a traditional enterprise option in which more data analysis could also aid the cannabis grower, by the customer purchases a license. The latter is typically identifying plant strains that are “good” versus those that may used for implementations in on-premise hardware or in have undergone mutations. 40 Tutorial

Holding Data to a Higher Standard, Part II: When Every Peak Counts—A Practical Guide to Reducing Contamination and Eliminating Error in the Analytical Laboratory

Operator, environmental, and method errors often include sources of contamination. In an increasing, more exacting analytical landscape in pursuit of parts-per-billion (ppb)-level analytes, it is very important not only to understand the sources of error and contamination but how to reduce them. During the dawn of analytical instruments, the laboratories tested for a select number of compounds or elements at parts-per-thousand levels. Modern instrumentation now has increased the number of compounds and elements to be quantitated and lowered the analytical threshold to sub-part-per-billion levels where 1 ppb is equivalent to 1 s in 32 years! In this type of testing environment even low parts-per- billion levels of contamination can cause large errors in quantitation. In this guide, we look at all of the most common sources of contamination and error in an analytical process from the water used in the laboratory to the inherent mistakes and error caused by laboratory equipment and operators.

Patricia Atkins

hemical reference standards have been an im- on how to best use CRMs. Sometimes a customer is not portant component in accurate analysis for dec- even aware that the issue they are questioning is really an Cades. Over the years, the challenges facing chem- problem of contamination or error. A common scenario ical laboratories have changed. As a manufacturer of is that a scientist will express a concern that their stand- certifi ed reference materials (CRMs), many questions ard is too high in particular elements. Usually during the

have arisen during our daily interaction with customers conversation, our scientists discover that the customer BIDAALA_STUDIO/SHUTTERSTOCK.COM

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is inadvertently allowing contamina- of contamination can dramatically solution and as a transfer solution for tion into their analysis. change laboratory values. volumetric or gravimetric calibrations Contamination and error can occur Most questions about contamina- or dilutions. In both of these uses, the at almost any point of the process and tion come in the form of an inquiry water must be clean to reduce con- then can be magnifi ed as the method about a particularly high result for tamination and introduce error into and analysis runs its course. Modern some common contaminant. In most the process. Poor quality water can instrumentation has raised the bar or cases, the root of that contamination cause a host of problems from creat- in this case lowered the limits of de- can be traced to a common source. ing deposits in labware or inadvert- tection to the parts-per-billion (ppb) The most common sources of stand- ently increasing a target element or or even parts-per-trillion (ppt) lev- ard and sample contamination are analyte concentration in solution. els. The concept of such small meas- found in the laboratory: reagents, The confusion starts when labora- urements would have been almost in- labware, laboratory environment, tories are unsure about which type of conceivable during the emergence of storage, and personnel. water they get from their water fi ltra- modern laboratory analysis. tion system. ASTM has guidelines that To put this into a different con- Water Quality designate different grades of water. text, 1 ppb expressed as a unit of time Water is one of the most basic yet Table I shows parameters for the four would be 1 s in 32 years and 1 ppt most essential laboratory compo- ASTM types of water (1). would be 1 s in 320 centuries! In the nents. Most scientists are aware that The actual type of water produced past, issues of laboratory contamina- the common perception that all water by a commercial laboratory water fi l- tion were problematic but now con- is the same is untrue. There are many tration system can vary in pH, sol- taminants, even in trace amounts, types, grades, and intended uses for utes, and soluble silica. Critical analyt- can severely alter results. It is hard water. Water is most often used in two ical processes should always require to imagine that such small amounts ways in the laboratory: as a cleaning a minimum quality of ASTM Type I

ADVANTAGE See What It Can Do for You and Your Lab

Pure Chromatography

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Table I: ASTM designations for reagent laboratory water (1)

ASTM Type Requirement I II III IV General lab grade Critical laboratory Cleaning glassware, used for pH, buffers, Use applications and feeding water polishing Not good for lab use feeding other water processes systems, water baths polishing systems Specific Resistance 18 1 4 0.2 (megohm/m) (max) pH N/A N/A N/A 5.0–8.0 Sodium (max) 1 μg/L 5 μg/L 10 μg/L 50 μg/L Total Silica (max) 3 μg/L 3 μg/L 500 μg/L high Total Organic 50 μg/L 50 μg/L 200 μg/L N/A Carbon (max)

their own water sources on an annu- Table II: Elemental leaching from laboratory tubing (μg/L) (2) al basis, but that does not mean it is Tubing Type Silicon Neoprene Silicon Neoprene applicable for use in laboratory applications. Municipal water can become Leaching Solution DI H2O DI H2O 5% HNO3 5% HNO3 Fe 0 0 27 5 contaminated from its distribution point, especially when left static sit- K2222 ting in pipes, tubing, and hoses. Wa- Mg 7 0.5 8 2 ter left stationary in a laboratory water Na 0 0.8 4 4 system can be exposed to leaching of Ni 0.2 0.2 4 0.1 elements and compounds from the Pb 0.1 0.1 3 2 piping and hoses. In one experiment Si 0 0 500 0 conducted at SPEX CertiPrep, leach- Sn 0.01 0.01 1 1.9 ing solutions of either ASTM Grade I Zn 1 55 4 50 water or 5% nitric acid in ASTM Grade I water were run through lengths of silicon and neoprene laboratory tubing. 91.0 The solutions were collected and test- 100 ed by inductively coupled plasma– 80 mass spectrometry (ICP-MS). Many common elements were found to have 60 leached into the solutions from expo- 28.7 40 sure to the laboratory tubing adding 9.1 potential contamination to critical an- 4.5 20 1.2 3.2 alytical processes, which can be seen in Table II. If the tubing was used to 0 HPLC LC–MS DI source DI source DI bottle Municipal transfer acidifi ed solutions between flushed stationary tap vessels the acid further increased the contamination, elements such as lead Figure 1: Phthalate concentrations in laboratory water sources (pbb) (3). increased from 0.1 μg/L in the ASTM I deionized (DI) water to 3 μg/L in the water. All trace analysis standards, di- standards, and samples with poor silicon tubing with 5% nitric acid and lutions, dissolutions, extractions, and quality water. 2 μg/L in the neoprene tubing with 5% digestions should be conducted with High-purity water is often achieved nitric acid. the highest purity of water. Analysts in several stages in multiple processes Another potential source of con- who use CRMs and perform quantita- that remove physically and chemically tamination of laboratory water is tive analysis need to use quality water potential contaminating substances. phthalates. Phthalates are ubiqui- so as not to contaminate their CRMs, Municipal water supplies often test tous in the environment and in the

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Table III: Grades of laboratory solvents and their uses

Grade Application or Use Analytical ACS General procedures Meets or exceeds ACS specifi cations Anhydrous Water-sensitive reactions and synthesis Low water levels (10–30 ppm) Biotechnical Biotechnical applications Low water, low residue, low UV Environmental analysis, Environmental Trace organics HPLC, trace organic Meets specifi cations of Food Food/FCC grade Food and drug applications Chemicals Codex (FCC) GC GC applications ppb levels of contamination Sub-micrometer fi ltration, some HPLC HPLC applications low UV absorbance LC–MS LC–MS applications Low ionic impurities, <0.1 ppm Pesticide, environmental analyses, trace Meets or exceeds ACS Pesticide residue analyses, and GC with various detectors pesticide specifi cations Reagent General laboratory use >95% purity Spectrophotometric UV applications UV, vis, IR Technical General laboratory use Noncritical tasks USP Food and drug applications Meets or exceeds USP specifi cations laboratory. Plastics are found every- Bottled-water sources can leach or- by individual manufacturers based where from bottles, tubing, caps, and ganic and inorganic contaminants on intended use. Some are general containers. In a laboratory water study from the bottle, cap, and liners. Bot- laboratory grades with intended use performed at SPEX CertiPrep, we ex- tled water can also have an expira- for noncritical applications while oth- amined bisphenol A and phthalate tion date and a shelf life that should er grades usually high in purity and content present in a variety of labora- be checked before use. Water that low in contaminants are designat- tory water sources including bottled has been decanted into other ves- ed for more-critical analyses. Table high performance liquid chromatog- sels can be exposed to many types III shows some general descriptions raphy (HPLC)-grade, liquid chroma- of contaminants including dust, mi- of types and grades of solvents and tography–mass spectrometry (LC– crobial growth, and oxidation effects. their intended uses. MS)-grade water, municipal tap water, Water left in liquid systems such as Many analytical laboratories use a and water obtained from our in-house HPLC, LC–MS, or ion chromatogra- variety of chromatography and spec- DI (ASTM I) water source (3). The in- phy (IC) systems should be changed trometry instruments such as gas house DI water was tested from a car- frequently to prevent contamination chromatography (GC), GC–MS, HPLC, boy fi lled within the laboratory, and and microbial growth. and LC–MS in their analyses. Solvents directly from the faucet outlet. The play a large role in these analytical faucet samples were collected after Reagents techniques as either mobile phases sitting overnight and a second sam- An analytical laboratory often uses or matrices for analysis. During sample was collected after the system had a large amount of various reagents, ple preparation, many laboratories been fl ushed with multiple gallons of solvents, and acids of varying quali- expose samples to a variety of sol- fresh water. Phthalates were found in ty and contamination levels. These vents ahead of introduction to an an- all the water sources with the high- chemical components can be a large alytical instrument. In some cases, the est total phthalates found in the HPLC economic investment for a laborato- kind of solvent is chosen to best fi t the bottled water with about 91 ppb of ry, but also a large source of poten- technique. LC, GC, and MS systems phthalates as seen in Figure 1. tial contamination. Just as in the case each have different modes of analysis To reduce contamination from lab- of water, there are different types or that benefi t from specifi c chemical or oratory water sources, the fi rst line of grades of chemicals, reagents, acids, physical properties found in solvents. attack is to choose the correct wa- and solvents. Some designations are For example, in GC and GC–MS, ter source for the given application. set forth by standards set by the U.S. the most widely used solvents have It is also important to realize that wa- Pharmacopeia (USP) or the American low boiling points, are eluted quick- ter can change quality over time de- Chemical Society (ACS). Other types ly, and don’t interfere with the target pending on the storage conditions. or grades of material are designated analytes. Solvents with boiling points

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acid, and nitric acid. Many of these ac- Table IV: Properties of common chromatography and spectroscopy solvents ids are commercially available in sev- Solvent MW UV Cutoff BP (°C) Polarity Index eral grades from general laboratory Water 18 NA 100 10.2 or reagent grade to high-purity trace Methanol 32 205 65 5.1 metal–grade acids. Acid grades often refl ect the number of sub-boiling dis- Acetonitrile 41.1 190 82 5.8 tillations the acid undergoes for puri- Acetone 58.1 330 56 5.1 fi cation before bottling. The more an 2-Propanol 60.1 205 82 3.9 acid is distilled, the higher the puri- Tetrahydrofuran 72.1 210 66 4 ty. These high-purity acids have the Ethyl ether 74.1 218 35 2.8 lowest amount of elemental contamination but can become very costly at DMSO 78.1 262 189 7.2 up to 10 times the cost of the reagent Cyclohexane 84.2 202 81 0.2 grade of acids. Methylene chloride 84.9 233 40 3.1 Often the question is asked if high Hexane 86.2 195 69 0.1 purity acids are necessary in sample Ethyl acetate 88.1 255 77 4.4 preparation if the laboratory is using Toluene 92.1 285 111 2.4 a high-quality ICP-MS-grade CRM. Clean acids used in sample prepara- Isooctane 114 205 99 0.1 tion, digestion, and preservation can be very costly. But, the difference be- within the analytical range of the tar- The solvents can also be contami- tween the amounts of contamination get analytes have the potential to be nated by gases, oxidation, or com- in a low-purity acid and a high-puri- coeluted with the targets and hinder pounds like the phthalates found in ty acid can be dramatic. High-quali- quantitation. (See Table IV.) seals and bottle closures. Some sol- ty standards for use in parts-per-bil- LC and MS techniques depend on vents have added preservatives that lion and parts-per-trillion analyses the type of analysis being performed. could add contaminants to analysis use the highest purity acids available Normal-phase LC uses polar columns or leach additional elements from to reduce all possible contamination and nonpolar solvents such as hexane the storage containers. from the acid source. An example of and cyclohexane. Reversed-phase LC There are many persistent sol- potential contamination is an aliquot uses nonpolar columns such as octa- vents that can be found in the labo- of 5 mL of acid containing 100 ppb decyl (C18) columns with polar sol- ratory which can cross-contaminate of Ni as contaminant, used for dilut- vents including methanol, water, samples by their presence. Some ing a sample to 100 mL, can introduce and acetonitrile. If ultraviolet–visible persistent solvents include dichlo- 5 ppb of Ni into the sample. light diode-array detection (UV-vis romethane which can cause chlo- To reduce contamination it is rec- DAD) or equivalent is used then the rine contamination as well as dime- ommended that high-purity acids be wavelength of the solvent also be- thyl sulfoxide (DMSO) and carbon used to dilute and prepare standards comes important. The wavelength of disulfi de, which can add sulfur resi- and samples when possible. In addi- the mobile phase and solvents used dues. There are solvents that react tion to using pure acid, it is important should be outside of the wavelengths with air to form peroxides, which can that the chemist check the acid’s cer- of the target analytes in the analysis. cause contamination and safety is- tifi cate of analysis to identify the ele- The most common range for a typi- sues in the laboratory. mental contamination levels present cal reverse phase is above 190 nm Acids are another laboratory re- in the acid. Some laboratories pre- UV cutoff for acetonitrile and below agent that can become both a po- fer to use blank subtractions to ne- 300 nm, which is the UV cutoff for ac- tential danger and a potential con- gate the background contamination, etone, both popular solvents for LC taminant. Acids by their nature are but blank subtraction for acids can applications seen in Table IV. oxidizers and many of the strongest only work in a range well over the in- Solvents are both a material that acids are used in the processing of strumental level of detection. If blank can become contaminated and samples for inorganic analysis. Com- subtraction causes an analytical result a source of contamination. Sol- mon acids in digestion and dissolu- to fall below the instrument’s level of vents can be contaminated by par- tion include perchloric acid, hydro- detection, it should not be used. ticles such as dust, rust, and mold. fl uoric acid, sulfuric acid, hydrochloric

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Table V: Dispensing volume error associated with various size syringes (4) Table VI: Compound carryover found in syringe washes (ppm of carryover) (4) Volume % of Syringe Syringe Size % Error Dispensed (μL) Volume Syringe Wash 1000 μL 10 μL Number 10 2 20% 23.15% 10 5 50% 8.16% 124.4360.51 10 10 100% 2.72% 20.6946.96 25 2 8% 8.82% 30.068.12 25 5 20% 5.47% 40.034.1 25 10 40% 2.37% 5 0.04 2.02 25 20 80% 1.05% 70.011.37 25 25 100% 1.25% 10 0.03 1.37 100 10 10% 6.09% 15 0.02 1.13 100 25 25% 1.67% 20 0.01 0.43 100 50 50% 0.64% 100 100 100% 0.61% laboratory from sample preparation to 1000 250 25% 1.05% standards calculation depend on ac- 1000 500 50% 1.14% curate and contamination-free volu- 1000 1000 100% 0.47% metric measurements. Unfortunately, laboratory volumetric labware, syringes, and pipettes are among the most Volumetrics and Labware repeated daily activity in most analyti- common sources of contamination, Volumetric measurement is a common cal laboratories. Many processes in the carryover, and error in the laboratory.

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liquid it will be marked by either the Table VII: Major elemental impurities found in laboratory container materials (2) letters TC or IN. Labware that is desig- Material Number of Elements Total ppm Major Impurities nated to deliver liquid will be marked Polystyrene-PS 8 4 Na, Ti, Al by either the letters TD or EX. Some- Tetrafl uoroethylene (TFE) 24 19 Ca, Pb, Fe, Cu times there are additional designa- Low density PE-LDPE 18 23 Ca, Cl, K, Ti, Zn tions such as wait time or delivery time Polycarbonate-PC 10 85 Cl, Br, Al inscribed on the labware. The deliv- Polymethyl pentene-PMP 14 178 Ca, Mg, Zn ery time refers to a period of time re- Fluorinated ethylene quired for the meniscus to fl ow from 25 241 K, Ca, Mg propylene (FEP) the upper volume mark to reach the Borosilicate glass 14 497 Si, B, Na lower volume mark. The wait time re- Polypropylene-PP 21 519 Cl, Mg, Ca fers to the time needed for the menis- High density PE-HDPE 22 654 Ca, Zn, Si cus to come to rest after the residual liquid has fi nished fl owing down from the wall of the pipette or vessel. Table VIII: Elemental impurities found in nitric acid distilled in clean laboratories versus regular laboratories (2) A second type of improper use and incorrect choice can be seen Element Regular Laboratory Clean Laboratory in the selection of pipettes and sy- Al 60 15 ringes for analytical measurements. As 0.17 <0.02 Many syringe manufacturers recom- Ca 150 100 mend a minimum dispensing vol- Cd 0.3 0.003 ume of approximately 10% of the to- Cr 2.5 0.4 tal volume of the syringe or pipette. Cu 1.7 0.23 A study by SPEX CertiPrep showed Fe 50 9 that dispensing such a small per- Mg 10 4 centage of the syringe’s total vol- Mn 1.1 0.1 ume created a large amount of er- Mo 0.8 0.03 ror. In this study, four syringes, 10 μL, Pb 0.5 0.4 25 μL, 100 μL, and 1000 μL were used Sb 0.04 0.013 to dispense between 8–100% of the Zn 5.5 0.7 syringe’s total volume of water. Each volume was weighed and replicated 10 times by several analysts and the The root of these errors is based on is designated as either Class A or results were averaged together to the four “I” errors of volumetrics: Class B labware. Class A glassware calculate average error. • Improper use is a higher quality analytical class The largest rates of error were seen • Incorrect choice of glassware whereas Class B glass- in the smaller syringes of 10 and 25 μL. • Inadequate cleaning ware is a lower quality glassware with Dispensing 20% of the 10 μL syringe • Infrequent calibration. a larger uncertainty and tolerance. created 23% error. Error only dropped These four I’s can lead to error and If a critical measurement process is down to below 5% as the volume dis- contamination, which negate all intent needed, then only Class A glassware pensed approached 100%. In the larg- of careful measurement processes. should be used for measurement. er syringes, measurements over 25% The fi rst two I’s stand for improp- Other information that can be were able to see error in and around er use, meaning that the volumet- found on labware is the name of the 1%. The larger syringes were able to ric is not used correctly or the incor- manufacturer, country of origin, tol- get closer to the 10% manufacturer’s rect choice is made. Many errors can erance or uncertainty of the meas- dispensing minimum without a large be avoided by understanding the urement of the labware, and a series amount of error, but the error did drop markings displayed on the volumet- of descriptors that indicate how the as the dispensed volume approached rics and choosing the proper tool glassware should be used. Labware 100%, which is seen in Table V. for the job. There is a lot of informa- can be marked with letters that des- The third “I” of volumetric error tion displayed on volumetric labware. ignate the purpose of the container. is inadequate cleaning. Many volu- Most labware, especially glassware, If a volumetric is designed to contain metrics can be subject to memory

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effects and carryover. In critical laboratory experiments, labware sometimes needs to be separated by purpose and use. Labware subject to Glass, Hair, Al, Cotton, Rubber, Ni, PS, tape, PVC, high levels of organic compounds or Air and skin wool, lead paper steel, Cu, S, PE, PP silicon, wood polyester PTFE persistent inorganic compounds can develop chemical interactions and Figure 2: Triboelectric charge potential of common materials and particles in the laboratory. memory effects. It is also sometimes diffi cult to eliminate carryover from labware and syringes even when using a manufacturer’s stated instruc- at high temperatures, used for corro- washer, and then the same aliquot of tions. For example, many syringes sive chemicals, or autoclaved should 5% nitric acid was drawn through the are cleaned by several repeated sol- be recalibrated more frequently. 5-mL pipettes. The aliquot was an- vent rinses before use. A study of sy- It is also suggested that under nor- alyzed by ICP-MS. The automated ringe carryover by SPEX CertiPrep mal conditions that soda-lime glass washer reduced the contamination showed that some syringes are sub- be checked or recalibrated every fi ve signifi cantly over manual cleaning of ject to high levels of chemical carryo- years and borosilicate glass after it the pipettes. The reduction of con- ver despite repeated rinses. has been in use for 10 years. The error tamination by moving from manual In this study, several syringes rang- associated with the use of volumet- cleaning to an automated cleaning ing in volume from 10 μL to 1000 μL rics can be greatly reduced by choos- process was clear. High levels of con- were used to dispense a 2000 μg/mL ing the correct volumetric for the task, tamination of sodium and calcium (al- internal standard mix of deuterated using the tool properly, and by mak- most 20 ppb) dropped to <0.01 ppb. compounds. The subsequent washes ing sure the volumetrics are proper- Other common contaminants includ- were collected and tested by GC–MS ly cleaned and calibrated before use. ing lead and iron dropped from 5.4 to determine the amount of carryover Inorganic analysts know that glass- and 1.6 ppb, respectively, to less in each wash (see Table VI). ware is a source of contamination. than 0.01 ppb. The larger syringes needed less Even clean glassware can contaminate The reduction of contamination in rinses to reduce carryover than the samples with elements such as boron, labware can depend on the material smaller 10-μL syringes. The smaller sy- silicon, and sodium. If glassware, such of the labware and its use. Different ringes had more than 1 ppm carryo- as pipettes and beakers, is reused, the materials contain many types of ele- ver through over 15 rinses. The typi- potential for contamination escalates. mental and organic potential contam- cal number of rinses usually employed A study was made of residual contamination as seen in Table VII (5). Trace in- to rinse syringes is between three and ination at SPEX CertiPrep of our pi- organic analyses are best performed fi ve, which in the case of the smaller sy- pettes after they were manually and in polymer or high purity quartz ves- ringe would not be adequate to clear automatically cleaned using a pipette sels, such as fl uorinated ethylene pro- all the carryover from the syringe. washer (2). pylene (FEP), and minimize contact The fi nal source of error is infre- An aliquot of 5% nitric acid was with borosilicate glass. Metals such quent calibration. Many laborato- drawn through a 5-mL pipette after as Pb and Cr are highly absorbed by ries have schedules of maintenance the pipette was manually cleaned ac- glass but not by plastics. On the oth- for equipment such as balanc- cording to standard procedures. The er hand, samples containing low levels es and automatic pipettes, but of- aliquots were analyzed by ICP-MS. of Hg (parts-per-billion levels) must be ten overlook calibration of reusa- The results showed that signifi cant re- stored in glass or fl uoropolymer be- ble burettes, pipettes, syringes, and sidual contamination still persisted in cause Hg vapors diffuse through pol- labware. Under most normal use, lab- the pipettes despite a thorough man- yethylene bottles. ware often does not need frequent ual cleaning procedure. calibration but there are some in- The experiment was repeated us- Laboratory stances where a schedule of recali- ing a pipette washer especially made Environment and Personnel bration should be used. Any glass- for use in parts-per-trillion analy- All laboratories believe they observe ware or labware in continuous use sis. The pipette washer repeatedly a level of laboratory cleanliness. Most for years should be checked for cal- forced deionized water through the chemists recognize that there are in- ibration. Glass manufactures suggest pipettes for a set time period. The herent levels of contamination pre- that any glassware used or cleaned pipettes were cleaned in the pipette sent in all laboratories. A common

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cosmetics. Even sweat and hair can cause elevated levels of sodium, calcium, potassium, lead, magnesium, and many ions. If a laboratory is seeing an

Cosmetics and Hair Medicated Soaps and perfumes products lotion cleaners usually high level of cadmium in the samples it could be from cigarettes, Metals, Phthalates, ZnO, Fragrances, dyes, colorants, pharmaceuticals, antibacterials, pigments, or batteries. If the levels of solvents, preservatives, waxes, alcohols metals fragrances, solvents lead are out of range, contamination VOA, SVOA can be from paint, cosmetics, and hair dyes. Figure 3 shows potential sources of common elemental contamination from outside products. Jewelry, Cigarette Lab Dust and all types smoke coats dirt Laboratory environment and per-

Rxns, plastics, VOA, SVOA, Everything in Pb, phthalates, sonnel contamination can be re- phthalates, paints, metals the lab! pesticides, VOA, dyes SVOA, heavy duced by limiting use of personal metals care products, jewelry, and cosmet- Figure 3: Common sources of contamination found in the laboratory. ics that could contain contamination and interfere with critical analyses. Laboratory coats can collect all types belief is that the small amounts of en- phthalates. The dust and rust particles of contamination and should only be vironmental and laboratory contam- can contaminate open containers in worn in the laboratory to avoid cross ination cannot truly change the ana- the laboratory or enter containers by contamination from other laborato- lytical results. To test the background charge transfer from friction by the tri- ries and the outside world. The labo- level of contamination in a typical lab- boelectric effect. The triboelectric ef- ratory surfaces should be kept clean. oratory, samples of nitric acid were fect or triboelectric charging is when Deionized water can be used to wipe distilled in both a regular laboratory materials become charged after com- down work surfaces. Laboratory hu- and in a clean-room laboratory with ing into contact with a second materi- midity can be kept above 50% to re- special air handling systems (HEPA al creating friction. The most common duce static charge. An ethanol- or fi lters). The nitric acid distilled in the example of this effect is seen when methanol-soaked laboratory wipe regular laboratory had high amounts hair sticks to a plastic comb after a can be used to reduce static electric- of aluminum, calcium, iron, sodium, static charge is created. ity as it evaporates. and magnesium contamination. Ta- The polarity and the strength of the Even with clean laboratory practic- ble VIII shows that the acid distilled in electrical charge is dependent upon es in place, erroneous results can of- the clean room had signifi cantly low- the type of material and other phys- ten fi nd their way into sample anal- er amounts of most contaminants (2). ical characteristics. Many materials in ysis. To eliminate some of these Laboratory air also can contrib- the laboratory have strong positive spurious results, replication of blanks ute to contamination of samples and or negative triboelectric charges as and sample dilutions can be used. standards. Common sources of air shown in Figure 2. In the laboratory, The blank results should be averaged and particulate matter contamina- materials like dust, air, skin, and lead and the sample run values can either tion are from surfaces and building have extreme positive charges and be minimally selected or averaged. materials such as ceiling tiles, paints, can be attracted to the strongly neg- The difference between the two val- cements, and dry wall. Surface con- ative charge of PTFE or other plastic ues can then be plotted against a taminants can be found in dust and bottles when the bottle is opened and curve established against two more rust on shelves, equipment, and fur- friction is created, inducing a charge. standards. A minimum of two stand- niture. Dust contains many different Laboratory personnel can add their ard points can be used if the chance Earth elements such as sodium, cal- own contamination from laborato- of contamination is minimal, such as cium, magnesium, manganese, sil- ry coats, makeup, perfume, and jew- in the case of rare or uncommon el- icon, aluminum, and titanium. Dust elry. Aluminum contamination can ements. Additional standard points can also contain elements of human come from laboratory glassware, cos- should be considered if the potential activities (Ni, Pb, Zn, Cu, As) and or- metics, and jewelry. Many other com- for contamination is high with com- ganic compounds like pesticides, per- mon elements can be brought in as mon elements such as aluminum, sistent organic pollutants (POP), and contamination from lotions, dyes, and sodium, and magnesium. Multiple

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2) SPEX CertiPrep Webinar, “Clean aliquots of blanks and dilutions can CRM manufacturers design stand- Laboratory Techniques,” https:// also be used to further minimize ana- ards for particular instruments to ob- www.spexcertiprep.com/webinar/ lytical uncertainty. tain the highest level of accuracy and clean-laboratory-techniques. performance for that technique. They 3) SPEX CertiPrep Application Note, General Principles and Practices also use calibrated balances, glass- “Analysis of Laboratory Water Laboratories should follow a gener- ware, and instruments to ensure the Sources for BPA and Phthalates,” https://www.spexcertiprep. al regime of three runs each of wash– most accurate standards are delivered com/knowledge-base/fi les/ rinse runs, blank runs, and sample to customers. Certifi cations such as AppNote_BPALabWater.pdf. runs, as well as single runs of sample ISO 9000, ISO 17025, and 17034 assure 4) SPEX CertiPrep Application Note, plus spike, and standard or spike runs customers that procedures are be- “Understanding Measurement: without sample to use as a control so- ing followed to ensure quality and ac- A Guide to Error, Contamination lution to evaluate recovery. curacy in those standards. After those and Carryover in Volumetric Analysts must realize that the clean- CRMs are in chemists’ hands, it is then Labware, Syringes and Pipettes,” available via the SPEX CertiPrep liness and accuracy of their proce- their responsibility to use all possible website as a downloadable PDF. dures, equipment, and dilutions af- practices to keep their analysis process 5) J.R. Moody and R. Lindstrom, fect the quality of the standards and free from contamination and error. Anal. Chem. 49, 2264 (1977). samples. Many laboratories will dilute CRMs to use across an array of References procedures and techniques. This in- 1) ASTM D1193-06(2018), Standard Patricia Atkins house dilution of CRMs can be a sav- Specifi cation for Reagent Water, is a Senior Applications Scientist with ings to the laboratory but in the fi nal (ASTM International, West SPEX CertiPrep in Metuchen, New Conshohocken, Pennsylvania, Jersey. Direct correspondence to: analysis can be a source of error and [email protected] 2018) www.astm.org contamination.

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Rapid Screening of Cannabinoids in Edibles Using Thermal Desorption-GC–MS

Most of the traditional methodologies for the determination of cannabinoids are based on solvent extraction, fi ltration, and concentration. These techniques are cumbersome, time-consuming, and suffer from analyst-to-analyst variability while producing data of limited value. Many laboratories routinely “screen” each sample to quickly determine the potential for matrix interference and instrument contamination while providing an estimate of the target compound’s concentration. A good “screening” method is simple (that is, minimal or no sample preparation), fast (analysis in less than 5 min), and semiquantitative. This article describes how thermal desorption (TD)-gas chromatography–mass spectrometry (GC–MS) analysis eliminates conventional sample preparation regimes and can be used as a good rapid screening technique.

Rojin Belganeh and William Pipkin

ost of the traditional methodologies for the de- compound’s concentration. A good “screening” method termination of cannabinoids are based on sol- is simple (that is, minimal or no sample preparation), fast Mvent extraction, fi ltration, and concentration. (analysis in less than 5 min), and semiquantitative. These techniques are cumbersome, time-consuming, One of the most widely used analytical techniques for and suffer from analyst-to-analyst variability while pro- “screening” is thermal desorption-gas chromatography– ducing data of limited value. mass spectrometry (TD-GC–MS) (1,2). This technique As the demand for the analysis of cannabinoids increas- does not require any solvent extraction or sample pre- es, it is imperative that the day-to-day analytical protocols treatment. Milligram quantities are put in an inert sam- be reproducible, accurate, and effi cient. Many labora- ple cup which is then “ready to analyze.” The multimode tories routinely “screen” each sample to quickly deter- pyrolyzer with a vertical micro-furnace design allows pro- mine the potential for matrix interference and instrument grammable and multiple thermal desorption analysis on

contamination while providing an estimate of the target a single sample. This process, which refers to evolved gas FUDIO/STOCK.ADOBE.COM

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analysis (EGA), starts with the acqui- sition of a thermal profi le (that is, de- 185 °C tector response as a function of sam- OH 4,000,000 H TIC ple temperature) of each sample 3,000,000 H O type. To perform EGA, a short, de- 2,000,000 activated tube (2.5 m, 0.15 mm i.d.) 1,000,000 is used to connect the injection port 200 300 400 500 600 700 to the MS detector. The sample is Thermal desorption range (100–300 °C) dropped into the furnace at a relatively low temperature (40–100 °C). m/z = 299 EIC The furnace is then programmed to m/z = 314 m/z = 231 a much higher temperature (600– 300,000 800 °C). Compounds “evolve” from m/z = 271 the sample as the temperature in- 200,000 creases. A plot of detector re- 100,000 sponse versus furnace temperature is obtained. Extracted ion chroma- 200 300 400 500 600 700 tograms (EIC) are used to identify the thermal zone over which specif- Figure 1: EGA thermogram of THC standard and extracted ion chromatograms (EICs). ic compounds of interest evolve from the sample. Now, these optimum TD temperatures can be used in subse- 8,000,000 Run #1 quent TD-GC–MS experiments to in- THC 0.26 mg troduce the key components of inter- 5,000,000 Area: 6083448 est while minimizing introduction of

the matrix. Only this portion of the 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 sample is actually transferred (that is, Run #2 8,000,000 injected on) to the analytical column. 0.26 mg Area: 6664705 Injecting only a small portion of the 5,000,000 sample provides immediate benefi ts to the laboratory, such as: 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 Run #3 • The high boiling fraction of the 8,000,000 0.25 mg sample remains in the sample 5,000,000 Area: 6352708 cup. This eliminates the need for a high-temperature bake

out. Thus, column lifetime 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 increases, there is little to no system contamination, and run- RSD 4.6% to-run cycle time decreases. • More sample can be put in the Figure 2: TD-GC–MS chromatograms of the brownie bite in triplicate. sample cup, which has the effect of lowering detection limits— instances, it is the reformed com- using the TD-GC–MS method for without affecting instrument pounds that are identifi ed and mon- quantifi cation of cannabinoids. The performance or cycle time. itored. Decarboxylation is forced experiments were performed by With respect to the analysis of can- to completion which could give the a Frontier Multi-Mode Pyrolyzer nabinoids, it is important to keep in “screening determinations” higher (EGA/PY 3030D) directly interfaced mind that TD-GC–MS is based on the values: the concentration range in- to a benchtop GC–MS system. To volatilization of target compounds creases and dilution factors are more automate the sequence and reduce from the matrix. Those compounds accurately determined. the workload by increasing reliabili- that are thermally labile or easi- ty of analysis, the Auto-Shot Sampler ly converted to an alternative com- Experimental (AS-1020E) was combined with the pound need to be identifi ed. In these Three edible samples were analyzed multimode pyrolyzer. The vent-free

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present in that brownie bite. The same brownie bite with the record- 1e+07 ed weight was used to perform the TD-GC–MS analysis to confirm the 5,000,000 theoretical THC value according to the label. To calculate and confirm the 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 amount of THC, EGA was first per-

16,000,000 formed on a THC standard (Abso- Add 300 ng THC y = 24,024.4071x + 6,222,377.7143 R2 = 0.9978 Add 100 ng 12,000,000 lute Standard). The pyrolyzer fur- CBN 8,000,000 Add 0 ng -259 nace was programmed from 100 °C CBD 4,000,000 to 800 °C (20 °C/min), and the EGA

0 -300 -200 -100 0 100 200 300 thermogram as shown in Figure 1 *STD solution is the mixture of CBD, THC and CBN Brownie Standard addition calibration curve was obtained. X intercept : -259.0 (ng) label THC From the EGA thermogram, the Sample weight 0.26 mg 10 mg/10 g= 1 mg/g THC concentration 996.2 ng/mg = 0.996 mg/g optimal thermal desorption zone of THC was identified as 100 °C to 300 °C. In fact, using the MS inter- Figure 3: Edible (brownie) sample standard addition calibration curve. pretation library, the peak with the apex of 185 °C (100 °C to 300 °C temperature zone) was identified as THC Run #1 THC. TD-GC–MS analysis was then 3,000,000 0.099 mg Area: 4460437 2,000,000 performed on the brownie bite in

1,000,000 triplicate as shown in Figure 2. To

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 perform TD-GC–MS analysis, the Run #2 pyrolyzer furnace was programmed 0.097 mg 3,000,000 Area: 4884453 from 100 °C to 300 °C (100 °C/min) 2,000,000 after the EGA tube was replaced by 1,000,000 a separation column (easily facili- 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 tated by using the vent-free GC– Run #3 0.105 mg MS adaptor). The amount of sample 3,000,000 Area: 4638695 used to obtain the TD chromato- 2,000,000

1,000,000 grams was 0.25–0.26 mg.

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 The peak shown in Figure 2 (not- ed with a *) between the retention RSD 4.6% time of 12 to 14 min was identified Figure 4: TD-GC–MS chromatograms of the chocolate pieces in triplicate. as THC using the MS interpretation library. The percent relative standard deviation (RSD%) of 4.6% was GC–MS adaptor that allows switch- cannabis-infused chocolate brown- calculated based on the area counts ing of a GC separation column with- ie containing 10 brownie bites with of the THC peak. The most intense out breaking vacuum on the MS de- the total of 100 mg tetrahydro- peak around 6 min was identified as tector was also used. The adaptor cannabinol (THC) was used. Ac- 5-hydroxymethylfurfural (dehydra- was utilized for switching the EGA cording to the product label, each tion of sugar). tube to the separation column and brownie bite contains 10 mg of To confirm the THC concentra- vice versa without venting the MS THC. In this experiment, one of tion in the brownie bite, a stand- detector, which saves time and in- the brownie bites from the packard addition calibration curve was creases productivity. age was placed on an analytical bal- created. The standard solution was ance, and the weight was recorded a mixture of cannabidiol (CBD), Sample 1 as 10.17 g. So, based on the label, THC, and cannabinol (CBN). Figure A commercial package of there is 10 mg/10 g = 1 mg/g of THC 3 shows the calibration curve and

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calculated amount of THC in the of each piece were recorded using the THC peak. brownie bite. an analytical balance as 0.099 mg, To confirm the THC concentration Using the calibration curve, the 0.097 mg, and 0.105 mg. in the chocolate pieces, a standard THC concentration is calculated as The same methodology was used addition calibration curve was cre- 0.996 mg/g while the product label for analyzing sample 2. First, the ated. The standard solution was indicates 1 mg/g of THC. EGA was performed as the fast rap- a mixture of CBD, THC, and CBN. id screening technique. Then the Figure 5 shows the calibration curve Sample 2 optimal thermal desorption zone and calculated amount of THC in A commercial cannabis-infused of THC was identified. The pyrolyz- the pieces. dark chocolate bar with the to- er furnace was programmed from Based on the calibration curve, tal of 100 mg THC and net weight 100 °C to 300 °C (100 °C/min) to ob- the THC concentration is calculated of 50 g (1.7 oz) was used for sam- tain the thermal desorption chro- as 2.1 mg/g while the product label ple 2. According to the product la- matograms for all three pieces of indicates 2 mg/g of THC. bel, there are 20 pieces of choco- the chocolate. Figure 4 shows the lates and each piece of chocolate TD chromatograms in triplicate. Sample 3 contains 5 mg of THC, so there is The peak shown in Figure 4 (not- A commercial cannabis-infused 100 mg/50 g = 2 mg/g of THC pre- ed with a *) between the retention gummy package with the net weight sent in each piece. To demonstrate time of 12 to 14 min was identified of 45 g (1.59 oz) was used for sample the accuracy and precision of the as THC using the MS interpretation 3. The package contains 10 gummies methodology, the analysis was per- library. The RSD% of 4.6% was cal- with THC and CBD (2-[(1R,6R)-6-iso- formed in triplicate. The weights culated based on the area counts of propenyl-3-methylcyclohex-2-en-

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To perform the TD-GC–MS analy-

THC 9,000,000 sis, one of the gummies was weighed 8,000,000 on an analytical balance The weight 7,000,000 6,000,000 was recorded as 0.109 mg. The sam- 5,000,000 4,000,000 ple was then placed in MeOH and 3,000,000 2,000,000 sonicated for 1 h. Next, 1 μL of 1,000,000 MeOH solution was spiked into the 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 sample cup.

Add 300 ng THC 14,000,000 It is important to mention that 12,000,000 y = 21,529.07x + 5,335,526.00 R2 = 0.994 Add 100 ng 10,000,000 the gummy sample could be ana- 8,000,000 Add 0 ng lyzed as is without dissolving in any -216.8 6,000,000 4,000,000 solvent. The experiment on sample 2,000,000

0 3 was performed based on exter- -300 -200 -100 0 100 200 300 *STD solution is the mixture of CBD, THC and CBN Brownie Standard addition calibration curve (TD300) nal standard calibration to demon- label THC X intercept : -216.8 (ng) strate the reliability and flexibility 100 mg/50 g= 2 mg/g Sample weight 0.105 mg THC concentration 2065 ng/mg = 2.1 mg/g of this technique. Figure 6 shows the TD-GC–MS Figure 5: Edible (chocolate pieces) sample standard addition calibration curve. chromatograms of the gummy samples in triplicate. The EGA thermogram of the gummy confirmed the thermal zone of 100 °C to 300 °C for CBD THC 1e+07 Area (m/z = 231):9530122 Area (m/z = 299):2823342 Run #1 both THC and CBD. Note the de- 6,000,000 0.25 mg tection peaks of both CBD and THC 2,000,000 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 in the chromatograms. The RSD% CBD THC Run #2 of 2.5% for CBD and 2.3% for THC 0.26 mg 1e+07 are calculated based on the area 6,000,000 9078163 2708138 2,000,000 counts of the THC peak. 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 CBD THC Run #3 To confirm the THC and CBD con- 0.26 mg 1e+07 centrations in the gummy sample,

6,000,000 9402197 2721218 an external standard calibration 2,000,000

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 curve was created. Figure 7 shows

RSD CBD 2.5% the calibration curves and the cal- RSD THC 2.3% culated concentrations, which are in excellent agreement with the Figure 6: TD-GC–MS chromatograms of the gummy in triplicate. package label. CBD concentration is calculated as 8.5 mg/gummy compared to the label value of Table I: Summary of the theoretical and calculated cannabinoids in the edibles 10 mg/gummy. The calculated THC Brownie Chocolate Bar Gummy is 6.1 mg/gummy while the label in- Target compound THC THC CBD THC dicates the value of 5 mg/gummy. Product label 1 mg/g 2 mg/g 10 mg 5 mg Calculated result 0.996 mg/g 2.1 mg/g 8.5 mg 6.1 mg Conclusion Table I shows the calculated and la- Repeatability 4.6 4.6 2.5 2.3 (N = 3) RSD (%) bel values of the cannabinoid concentrations in all three commercial Calibration 0.998 0.994 0.999 0.998 curve (R2) edible samples. Using TD-GC–MS the cannabinoids were thermally ex- 1-yl]-5-pentylbenzene-1,3-diol). Ac- tetrahydrocannabinolic acid (THCA), tracted from the mixture and the cal- cording to the product label, each cannabidiolic acid (CBDA), and CBN culated concentrations were in ex- gummy has 10 mg CBD and 5 mg (other required cannabinoids) are cellent agreement with the package THC. The label also indicates that 0.0 mg/package. labels. Standard addition calibration

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curves were used for the brownie bite and chocolate piece while an ex- THC 50 mg/package CBD 100 mg/ ternal standard calibration curve was created to calculate THC and CBD in (CBN) 1000 ng 1.5e+07 the gummy sample. THC THC 50 mg/package CBD 100 mg/package 1e+07 CBD In conclusion, thermal desorp- (THC 50 mg/gummy, CBD 10 mg/gummy) 5,000,000 300 ng tion analysis eliminates convention- 100 ng 11.00 11.50 12.00 12.50 13.00 13.50 14.00 14.50 15.00 al sample preparation regimes; the CBD THC sample is heated to the point that 40,000,00 20,000,00 y = 38389x - 2E+06 y = 17202x - 680172 the cannabinoids desorb from the 30,000,00 R2 = 0.99869 15,000,00 R2 = 0.99825 edible matrix (thermal extraction). 20,000,00 10,000,00 10,000,00 It is fast, uses minimal or no solvent, 5,000,00 0 and eliminates the need for expen- 0 200 400 600 800 1000 1200 0 0 200 400 600 800 1000 1200 sive glassware. TD-GC–MS is a “vol- CBN atiles only” analysis; high boiling 100,000,000 80,000,000 y = 86574x - 3E+06 CBD THC R2 = 0.99907 sample constituents remain in the 60,000,000 Label 10 mg/gummy 5 mg/gummy sample cup which eliminates sys- 40,000,000 20,000,000 Calculated 8.5 mg/gummy 6.1 mg/gummy tem contamination, increases sys- 0 (ESTD) 0 200 400 600 800 1000 tem stability, and reduces run-to-run 1200 analysis time. A multimode pyrolyzer provides Figure 7: Edible (gummy) sample external standard addition calibration curve. users with a clear picture of the sample’s composition by identifying the thermal zones and the com- complex polymers, copolymers, vol- References pounds in each zone. Using the atiles, and additives as well as failure, 1) ASTM International, ASTM D7823- EGA thermogram, one can simply contamination, deformulation, and 16: Determination of Low Level, determine the suitable temperature degradation analyses. As opposed Regulated Phthalates in Poly (Vinyl program and program the pyrolyz- to GC–MS alone, pyrolysis GC–MS Chloride) Plastics by Thermal Desorption—Gas Chromatography/ er’s furnace appropriately. allows multiple and customized anal- Mass Spectrometry, https://www. Using pyrolysis GC–MS, solid sam- ysis on the same sample while the MS astm.org/Standards/D7823.htm. ples can be analyzed. There is no libraries offer a broad range of refer- 2) International Electrotechnical solvent required when using pyrol- ences: from low boiling point com- Commission, IEC 62321-8: ysis-thermal desorption GC–MS as pounds to pyrolyzates and heavier Determination of certain substances opposed to traditional GC–MS tech- polymeric fragmentations. In terms in electrotechnical products – Part 8: Phthalates in polymers by gas niques. In other words, the solid and of the ability to run samples at high- chromatography-mass spectrometry liquid samples can be injected (user temperature, pyrolysis GC–MS (GC-MS), gas chromatography-mass ing an inert sample cup) into the py- can provide the user with an expand- spectrometry using a pyrolyzer/ rolyzer without any solvent and sam- ed temperature range from ambient thermal desorption accessory (PY/ TD-GC-MS), https://webstore. ple pretreatment. This advantage, as +10 to 1050 °C (± 0.1 °C precision) as iec.ch/publication/32719. well as rapid screening capability, is well as a high interface temperature one of the primary reasons many lab- (maximum 450 °C). oratories integrate multimode py- rolyzers into both their day-to-day Acknowledgement quality control and analytical re- The authors thank Robert Free- search protocols. man, Itsuko Iwai (Research & Devel- Rojin Belganeh is the technical Pyrolysis GC–MS techniques also opment, Frontier Laboratories), and and marketing director for Frontier Laboratories Ltd. North America in enable material characterization of Terry Ramus (application scientist, Antioch, California. William Pipkin virtually any organic material by pro- Diablo Analytical) for contributing to is the president of ATRq in Orem, viding detailed information about the article and providing the analyt- Utah. Direct correspondence to: [email protected] and william. the composition of the samples. This ical work, without which this article [email protected] technique is used for identifi cation of would not have been possible.

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Major Engine of Growth in the Cannabis Industry: State of the Art Formulation Technology for Product Innovation and Expanded Applications

Formulation technologies provide a powerful tool in product innovation and new applications especially for purifi ed and highly concentrated extracts including cannabidiol (CBD) and tetrahydrocannabinol (THC). Fortunately, the cannabis industry can choose from formulation technologies and ingredients that have a record of use in pharmaceuticals, food and beverages, dietary supplements, personal care, and veterinary and pet food products. This article aims to assist in making choices that support safety, effi cacy, stability, and consumer acceptance.

Andreas M. Papas

ew, innovative products and applications have (GW Pharmaceuticals), which is an oral solution of canna- been a major driver of growth of the cannabis in- bidiol (CBD) for the treatment of seizures associated with Ndustry. The range and number of new products two rare and severe forms of epilepsy. Epidiolex is the fi rst is breathtaking and spans pharmaceuticals, food and di- FDA-approved drug that contains a purifi ed drug sub- etary supplements, topical and personal care, and veter- stance derived from cannabis—in a strong pipeline of de- inary and pet food. It includes product formats ranging velopment and clinical testing. from prescription pharmaceutical formulations to capsules, Purifi ed and concentrated cannabis extracts are power- chewable tablets, nutrition bars, gummies, drops, bottled ful tools in product innovation. For medical applications, water, beverages and sports drinks, creams, lotions, sprays, they allow the identifi cation of the active ingredients, clin- inhalation and vapor products, and pet snacks and treats. ical testing for FDA approval and claims, and products In a signifi cant milestone, in June 2018 the U.S. Food and that have defi ned potency and can be tested with estab-

Drug Administration (FDA) approved the drug Epidiolex lished scientifi c methods for meeting specifi cations with WITTHAYA/STOCK.ADOBE.COM

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consistency and stability. The same • Taste, odor and other health conditions of the user. Clinical- reasons increasingly apply to dietary characteristics ly proven formulation technologies in- supplements, beverages and sports • Regulatory crease absorption and bioavailability drinks, and other applications. The Let’s take a closer look at each reason especially in the large population seg- use of purifi ed and concentrated ex- below. ments with suboptimal absorption (1,2). tracts also allows for the use of smaller amounts, which has signifi cant ben- Solubility in Water Improved Stability efi ts for taste, odor, color, or capsule Upon extraction and purifi cation, im- The stability and shelf-life of can- and tablet size for oral supplements. portant cannabinoids such as CBD and nabinoids is affected by exposure The use of purifi ed and concentrate tetrahydrocannabinol (THC) come out to sunlight, air, heat, and other fac- extracts also facilitates accurate test- as oils that are practically insoluble in tors. When cannabinoids are adding using smaller sample size. water. For many products and applica- ed to water, foods, creams, and oth- tions such as beverages, medical for- er products, their stability can be Why Formulation: mulations, sprays, topical formulations, affected by exposure to oxidizing The Top Reasons and others, solubility in water is a must. agents, acidic or alkali conditions, Formulation is key to new innovative and reactive compounds. Custom- products and applications for the fol- Increased Absorption ized formulations provide powerful lowing reasons: and Bioavailability tools for optimizing stability. • Solubility in water Absorption of oils requires optimal • Increased absorption function of the liver, pancreas, and the Taste, Odor, and and bioavailability complete digestive tract. Absorption Other Characteristics • Improved stability is also affected by the diet, age, and Taste is of paramount importance

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parameters of water solubility, stability, and increased absorption and bioavail- Intestine ability. While effi cacy claims are abun- Micelles dant, few are supported by evidence. The gold standard is research and clin- Lipids Blood ical evidence from leading universities and researchers, preferably support- Pancreatic Bile ed by the National Institutes of Health enzymes (NIH) or similar quality funding organizations, and the results published in ref- ereed scientifi c and medical journals. Pancreas Liver Positive or Neutral Effect on Important Properties Figure 1: Absorption of lipids in humans. Bile and pancreatic enzymes are essential for This includes taste, odor, and color. the formation of micelles and absorption of lipids. Suboptimal function and diseases of For oral products, especially foods the liver, pancreas, and the digestive tract cause malabsorption. and beverages, taste and odor are of paramount importance.

Support of Label Lipophilic interior and Customer Preferences Hydrophilic exterior Although there is no offi cial defi nition of clean label, consumers increasingly show preference for minimally processed products with safe ingredients, preferably derived from natural Figure 2: Micelle-like particles enable solubility and enhance absorption. Micelles are sources. Additional consumer prefer- composed of hydrophobic (lipid) interior and hydrophilic exterior. In the human body, ences include no sugar or salt added, formation of micelles is essential for emulsifi cation and absorption of lipids. free of gluten, dairy, and animal products, nuts and other sources of aller- in food, beverages, and other oral Choosing Formulation gens, and nongenetically modifi ed products. Odor is also important. Agents: Top Criteria organisms (non-GMO). Terpenes, which are abundant in can- The top criteria to consider when nabis plants, can have strong aro- choosing the right formation agents Key Commercial mas that may be carried over into include safety, effi cacy, positive or Formulation Applications extracts, concentrates, and even pu- neutral effect on important proper- The major cannabinoids, CBD and rifi ed cannabinoids. Formulation pro- ties, and support of label and cus- THC, are key components of innova- vides the opportunity for address- tomer preferences. Let’s explore each tive products. Increasingly, these can- ing the taste, odor, texture, and other of those criteria in more detail. nabinoids are used in concentrated or characteristics for the specifi c prod- purifi ed form, especially for medical, ucts and applications. Safety dietary supplement and food and bev- The FDA’s GRAS (generally recog- erage applications, sprays, and topical Regulatory nized as safe) designation is a top se- products. Key objectives of formulation For medical applications, formulation lection criterion. A long record of safe were discussed above and include: is required for accurate dosing of the use on a daily basis, with signifi cant desired product. With the expand- safety margins across wide dose rang- 1. Solubility in water especially ing approval of cannabis products in es supported by clinical evidence, for the emerging market the United States, Canada, and oth- provides additional confi dence. segments of infused water, er countries, there is increasing focus beverages, sports drinks, and on quality, testing and product speci- Effi cacy other water-based products. fi cations, potency, stability, and other This criterion requires major home- 2. Increased absorption and product characteristics. work, especially for the critical bioavailability, which is key

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of allergens, sugar, salt, gluten, for medical applications with glycols, and glycerin support the for- dairy, and animal products. oral dosing especially because mulation of liquids. disease conditions often cause The increasing consumer aware- References poor absorption. Enhanced ness and strong preference for simple 1) S.H. Wu and W.K. Hopkins, Pharm. absorption is important for formulations using ingredients with Technol. 23, 52–68 (1999). effi cacy of dietary supplement proven records of safety and effi cacy, 2) C. Högenauer and H.F. Hammer, applications in wellness and and the major role of taste and odor, in Sleisenger and Fordtran’s health with strong and growing especially for oral products, have lim- Gastrointestinal and Liver Disease, consumer awareness ited the practical choices dramatical- 10th Ed., M. Feldman, L.S. Friedman, and L.J. Brandt, Eds. 3. Stability: Formulations of ly. For example, polysorbates (such as (Elsevier Saunders, Philadelphia, cannabinoids in water based Tween-80) which are common, strong Pennsylvania, 2016). Ch. 104. products, especially when solubilizers in the pharmaceutical, 3) L.K.Sørensen and J.B. Hasselstrøm, combined with other ingredients food and beverage, and cosmetic in- Drug Test Anal. 10, 301–309 (2018). in the fi nal product such dustries, can impact the taste. More 4) http://www.ewg.org/skindeep/ as vitamins, minerals, and importantly, an increasing consumer ingredient.php?ingred06=705142& nutraceutical, increase the segment prefer products that do not refurl=%2Fproduct.php%3Fprod_ challenges to their stability (3). contain polysorbates (4). id%3D62813%26#.W6T0dvZRfyQ. Vitamin E TPGS, a derivative of nat- 5) European Food Safety Authority A good number of solubilizing agents urally sourced alpha-tocopherol is (EFSA), “D-alpha-tocopheryl are available and include: becoming the formulation ingredient polyethylene glycol 1000 succinate • Water-soluble organic solvents of choice for cannabis products, es- (TPGS) in use for food for particular such as ethanol, polyethylene nutritional purpose,” The EFSA pecially purifi ed or concentrated CBD Journal 490, 1–20 (2007). glycols, glycerin, N-methyl-2- and THC and other extracts. The rea- pyrrolidone, dimethylacetamide, 6) K.A. Papas, M.K. Sontag, C. sons include: and dimethylsulfoxide (DMSO). Pardee, R.J. Sokol, S.D. Sagel, • A proven record of safety (1,5) F.J. Accurso, and J.S. Wagener, • Non-ionic surfactants such as based on decades of commercial J. Cyst. Fibros. 7, 60–67 (2008). vitamin E TPGS (d-α-tocopheryl use in pharmaceuticals and 7) P.P. Constantinides, J. Han, polyethylene glycol 1000 dietary supplements, including and S.S. Davis, Pharm. succinate), polyethoxylated vitamin formulations for children. Res. 23, 243–55 (2006). castor oil (Cremophors), polysorbates (Tweens), sorbitan • Effi cacy based on conclusive monooleate, poloxamers, clinical evidence (6), not only macrogolglycerides linoleates, and as a solubilizer for water- Andreas M. Papas, PhD, is CEO and member of the Board of Directors of mono- and di-fatty acid esters. soluble formulations, but also in its ability to form micelle- Antares Health Products, Inc., and an • Phospholipids such like particles that enhance Adjunct Professor of Health Sciences of East Tennessee State University. A as hydrogenated soy absorption and bioavailability. phosphatidylcholine, Fulbright Scholar, Dr. Papas is a grad- Its effi cacy is illustrated by its uate of the University of Illinois and distearoylphosphatidylglycerol, use in clinically-tested (7), FDA- α author of The Vitamin E Factor paper- l- -dimyristoylphosphatidylcholine, approved products formulated back and editor of the scientific book α l- -dimyristoylphosp for enhanced absorption. Antioxidant Status, Diet, Nutrition and hatidylglycerol. Health. Dr Papas also founded YASOO • Organic liquids and semi-solids • Neutral taste and odor and light Health Inc., and led the company as such as medium-chain mono- and yellow color are particularly President and Chair of the Board of diglycerides, and cyclodextrins. important for oral and personal Directors. He developed product concepts and managed successful care product formulations. Producing the commercialization including formula- Optimal Formulation • Support of product stability. tion, clinical evaluation supported by the National Institutes of Health and The choice of the right ingredients Vitamin E TPGS contains some the Cystic Fibrosis Foundation, stabil- depends on many factors such as the free tocopherols that have strong ity and safety testing, as well as pilot antioxidant activity and support properties of the active ingredient or and commercial production and mar- product stability especially for keting. These disease specific nutri- the fi nal form of the product. For ex- oil-based products. In addition tional products are considered the ample, cyclodextrins and solid mono- to reducing potency, products standard of nutritional care in cystic glycerides have a key role in formulat- of oxidation affect taste, fibrosis, liver disease, and bariatric patients. Direct correspondence to: odor, and can be harmful. ing tablets and other solid forms while [email protected] others such as ethanol, polyethylene • Support of label claim of free

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Professor Jack Henion Shares Insights on Cannabis Research, Academia, and the Expanding Role of Mass Spectrometry Cannabis analytical testing is rapidly expanding beyond cannabis quality control testing to academic, pharmaceutical, and clinical markets. Mass spectrometry is also playing a critical role in this evolution. In this month’s “Cannabis Crossroads,” Professor Jack Henion shares his insights on cannabis research, academia, and mass spectrometry, including challenges and opportunities in moving cannabis science forward.

Joshua Crossney

It seems most analytical scientists high-quality analytical techniques and some advancement, but the technolo- have an “ah ha!” moment when they technology. I remain excited about gies and methods available when prop- realize the incredible opportunities working within and contributing to this erly applied are not that diffi cult. Of in cannabis science. When did you fast-paced, evolving industry. course, there are additional quantita- start to see these opportunities, tive measurements needed such as for and how did you fi rst get involved in Every scientist that I meet with terpenes, fl avonoids, mycotoxins, and cannabis science? struggles in some way with analyzing heavy metals, which in some cases re- Professor Jack Henion: I actually had cannabis. What hurdles did you fi rst quire different sample preparation tech- two “ah ha” moments. The fi rst was as experience in working with cannabis niques and instrumentation. Thus, a “to- an analytical chemist I was intrigued to and how did you overcome them? tal analysis” of a cannabis plant sample see how analytical methodologies and Henion: Our experience with marijuana is not just one sample preparation tech- technologies used for cannabis were plant materials to-date is somewhat limit- nique followed by an injection into one not as technologically current as those ed because we do not yet have a Sched- analytical system. It is a series of related, in the good laboratory practice-reg- ule I controlled substances license from but different procedures. ulated pharmaceutical bioanalytical New York State. We have applied for this world with which I was familiar. This cou- license, but do not yet have it. As a result, You have started lecturing on pled with the chemical complexity and I have focused on the analysis of hemp cannabis and hemp at Cornell diversity of the many cannabis cultivars plant materials and products which of University. What were some of your and the importance of accurate and re- course must contain less than 0.3% tet- takeaway messages and how were liable analyses suggested that perhaps rahydrocannabinol (THC). I would not those lectures received? I could contribute to improving the an- say that the analysis of these plant ma- Henion: I recently gave a lecture to the alytical sciences used in this growing in- terials is particularly diffi cult. In contrast Cornell BioEngineering MS graduate dustry. However, the real excitement, to my experience with parts-per-billion students where I focused on the impor- or second “ah ha” moment for me, oc- (ppb) and sub-ppb bioanalysis of drugs tance and benefi ts of collaboration and curred when I attended the fi rst Canna- and their metabolites in biological fl u- innovation in science and business de- bis Science Conference in 2016 where ids, quantifying the important cannabi- velopment. I described the rapidly de- I witnessed compelling testimonials noids in hemp and marijuana is relatively veloping cannabis industry as a new fron- of the very positive medicinal marijua- easy because the levels are much high- tier with many technology development na benefi ts afforded to certain disease er (percent levels) and the matrix is much opportunities for these students in their sufferers. A common successful busi- less complex than blood or urine. In con- bioengineering careers. I received many ness model is to “fi nd a need and fi ll it.” trast, the accurate and precise quanti- inbound inquiries after this lecture, which I was convinced this was an exciting new tative determination of as many as 100 suggests I struck a chord of interest and fi eld where I could perhaps contribute pesticides at low ppb levels in marijua- excitement with these students by shar- to “fi lling a need” by providing positive na plant materials is of course more chal- ing with them some industry intellect re- contributions to supporting the scientif- lenging. Plant sample preparation when garding opportunities in the cannabis

ic integrity of this new industry through handling large sample numbers needs and hemp industries. OLEH MARKOV/SHUTTERSTOCK.COM

CANNABIS SCIENCE AND TECHNOLOGY | www.CannabisScienceTech.com VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 Cannaabis Crossroaads 61

Additionally, I recently presented two cannabis potency determinations be- plot on my personal farm near Ithaca, NY. lectures at the annual Cornell Veterinary cause we know there are chemical en- So, now I know a little bit about growing Conference at the Cornell University Col- tities in the sample matrix that can coe- and analyzing hemp. lege of Veterinary Medicine, Ithaca, NY lute or interfere with photodiode-array (1). There is growing interest and accept- detection. If LC–MS techniques could be Are there any additional comments ance, especially among companion an- competitively priced and coupled with you would like to make regarding the imal veterinarians for the use of canna- ease-of-use, I would prefer to use those future of cannabis science? bidiol (CBD) oils and related products techniques for potency determinations Henion: I believe medicinal marijuana to manage pain and other ailments for rather than LC–PAD. But that should be and its promising industry will be part of their “patients.” These lectures included left up to the individual laboratory and its the future. However, we have a very steep a “tutorial” on liquid chromatography– leadership. learning curve ahead of us. Academic re- mass spectrometry (LC–MS) and related search is thwarted because of misguid- techniques showing methods used for Please comment on what is next for ed opinions and funding is limited for the analysis of hemp and marijuana plant you in cannabis science. What business similar reasons. The good news is, I be- materials as well as a second lecture on and research projects are you working lieve, this will change with time and that the comparison of analytical results from on that you can share with us? in fact there are many exciting opportu- commercial veterinary oils, tinctures, and Henion: As you may perceive from my nities ahead. My plea is for the mainte- so forth versus what is listed on the labels. comments above, I am very excited to nance of scientifi c and business integrity My message was caveat emptor since of- participate in and hopefully contrib- such that the benefi ts and opportunities ten the product label does not accurately ute to maintaining a high degree of sci- remain available for all. As Nike says, let’s refl ect what is in the bottle. The veterinar- entifi c excellence used in the analysis of “just do it.” ians in the audience appeared to be very any cannabis-related samples. This per- interested in these topics. tains to plant materials for potency and Reference pesticides as well as the rapidly increas- 1) http://www.cvent.com/events/2018- As a pioneer and leader in mass ing diverse array of commercial prod- fall-new-york-state-veterinary- spectrometry, can you please comment ucts. I have heard reference to the “wild, conference/agenda-7c9177edd8 on its role in cannabis science? wild west” nature of some current pro- 2241ccb90849e19093c5a3.aspx. Henion: I believe mass spectrometry is cedures and policies in the medical mar- an essential component for the credi- ijuana world. The days of “snake oil” About the Interviewee ble growth and acceptance of the can- sales pitches and unsubstantiated prod- Dr. Jack Henion is an nabis industry. I, of course, can be ac- uct promises should not return because internationally recognized cused of being “overly prejudiced” in my it will diminish the credibility and poten- leader in the field of MS and interest and support of MS as a detector tial of this exciting fi eld. I have proposed LC–MS. He is credited with 13 patents and has pub- and, in particular, LC–MS techniques. In that Advion, Inc., establish a “Cannabis lished more than 200 peer- my early days of racehorse drug testing CRO” and plans for that are underway. reviewed papers in scientific journals. In where our analytical results would have to It is my goal to establish and maintain a addition to vast industrial experience, Dr. Henion managed a major research stand up to legal scrutiny in court, my stu- state-of-the-art laboratory that maintains laboratory at Cornell University, where dents used to call the mass spectrome- the highest level of scientifi c analytical he served as a professor of toxicology for ter the “truth machine.” Currently, there services. In the meantime, we are carry- more than 24 years and is now Emeritus Professor of Analytical Toxicology. is no other detector for the determina- ing out applied research in sample prep- tion of organic compounds that provides aration and LC–MS analysis techniques the sensitivity, selectivity, and speed for for hemp-based plants and products About the Columnist the money than MS. When MS is cou- to improve upon analytical procedures Joshua Crossney is the pled to gas chromatography (GC) and currently used. To have a ready source columnist and editor of for even wider applications, LC (or best of fresh hemp plant materials, this past “Cannabis Crossroads” and a contributing editor with ultrahigh-pressure liquid chroma- summer I qualifi ed as an approved “Cor- to Cannabis Science and tography [UHPLC]), one has unequaled nell Affi liate” hemp grower under a per- Technology magazine. analytical capabilities. I worry a bit about mit issued by the New York State Agricul- Crossney is also the president and CEO of CSC Events. Direct correspon- the current unchallenged interest in LC– ture and Markets and collaborated with dence to: [email protected]

photodiode-array detector (PAD) for Cornell University to grow a small hemp OLEH MARKOV/SHUTTERSTOCK.COM

VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 www.CannabisScienceTech.com | CANNABIS SCIENCE AND TECHNOLOGY 62 Applicationn Note ADDVERTISEEMENT

The Advantages of an Industry-Specifi c LIMS for Cannabis Labs

Marty Pittman, Senior Product Manager, LabVantage Solutions Inc.

LabVantage Cannabis is a purpose-built informatics platform, offering cannabis testing labs approximately 80% of commonly required industry functionality.

People in industries that have long been using laborato- Tracking Capabilities ry information management systems (LIMS) remember the Both growers and lab administrators as well as lab man- days (or months or years) of customizing a base software agers want visibility to lab processing. Lab leaders must platform into a workable solution specifi cally for their or- be able to drill down into the details of particular re- ganization. The emerging cannabis industry is now bene- quests, and automatically send any questions back to fi tting from these years of customization, as LIMS vendors growers. Password-protected electronic signatures are have begun building more of the common, necessary func- required throughout the process to maintain a digitized tionality into the base product. chain of custody. Industry accelerators take this approach a step further; the LIMS platform is purpose-built for cannabis testing labs, METRC Integration offering right out-of-the-box approximately 80% of com- As samples are received and processed, the LIMS should monly required functionality and, for the remaining 20%, interface with METRC to signal that the sample is under- it’s easily confi gured without the need for coding. The big- going testing. Likewise, the system should share results gest advantage of purpose-built software is the speed of and auto-populate invoices. Such integration contributes deployment, and the subsequent reduced cost and risk. to a more effi cient workfl ow between the grower, the lab, Industry accelerators can cut implementation time by as and regulators. much as 75%, and savings by about the same. Without the need to spend time analyzing workfl ows and Industry-Standard Testing processes to determine system requirements, the acceler- A cannabis LIMS must support industry-standard testing, ated LIMS gets right to helping reinforce consistent, indus- such as American Herbal Pharmacopoeia-recognized ana- try-standard workfl ows that lead to greater effi ciency and lytical test methods. productivity. LIMS prepackaged with electronic lab note- A purpose-built cannabis LIMS works in lockstep with book functionality and industry-specifi c modules create a lab teams, from initial request through fi nal invoicing, in a streamlined system that maximizes return on investment way and at a deployment speed that traditional LIMS can- while lowering risk because custom code is not needed. not. The industry-accelerated LIMS enables a highly effi - With an interface that is responsive to the specifi c industry, cient and scalable workfl ow supported by industry-stand- the intuitive purpose-built LIMS reduces the time and in- ard quality control. vestment in user training. For a white paper and information on LabVantage Can- nabis, a cannabis-specifi c LIMS platform, visit labvan- What to Look for in an Industry-Specifi c LIMS tage.com/cannabis. Grower Portal Because cannabis testing labs serve a wide range of growers, a portal into the system is a desirable feature. It provides growers with a frictionless mechanism for preparing and submitting requests for the lab. Growers must be able to easily enter details about the LabVantage Solutions, Inc. sample; specify which tests are needed and when; provide 265 Davidson Avenue, Suite 220, details such as METRC ID or the mass of each batch; and Somerset, NJ 08873 track all of their harvest lot requests at various stages. The tel. (908) 707-4100; fax (732) 560-0121 system must deliver a certifi cate of analysis that is modifi a- website: www.labvantage.com ble for different lab results.

CANNABIS SCIENCE AND TECHNOLOGY | www.CannabisScienceTech.com VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 MANUFACTURING & PROCESSING INSTRUMENTS PACKAGING & CULTIVATION/ EXTRACTION CONSULTING TESTING LAB LAB TESTING ANALYTICAL ANALYTICAL GROWING SERVICES SERVICES SUPPLIES OTHER OTHER

Supplier Profi 2018 ISSUE les

Evgeniy - stock.adobe.comobe.com 64 Supplier Proﬁ le

Advion & Advion BioServices COMPANY DESCRIPTION ANALYTICAL Advion’s nearly three-decade dedication to serving scientists yields customer-focused life sci- INSTRUMENTS ence solutions. Our deep scientifi c, engineering, and customer workfl ow knowledge spawns an unrivaled solution portfolio. We work directly with, train, and passionately advocate for our customers to ensure their success. Dedicated to Science—Dedicated to You.

CHIEF SERVICES SUPPORTED TERRITORIES SERVED ANALYTICAL Advion develops, manufactures, and global- Global instrumentation provider with contract TESTING LAB ly supports mass spectrometers, novel sam- lab services for US and Canadian markets. ple introduction and assay solutions, chip- based ion sources, chromatography systems, and consumables for life science and related industries. Advion also offers expanded full- service contract research testing facilities for cannabis, hemp, and biological samples.

MAJOR PRODUCTS • expression compact mass spectrometer • SOLATION ICP-MS • Plate Express TLC plate reader • AVANT (U)HPLC chromatography series • TriVersa NanoMate nanoelectrospray ionization source

Advion & Advion BioServices

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CEM Corporation COMPANY DESCRIPTION ANALYTICAL CEM Corporation is a leading global company specializing in scientifi c solutions for critical lab- INSTRUMENTS oratory applications. We design and manufacture systems for analytical laboratories, life sciences, and processing plants worldwide. Our product portfolio includes innovative instrumentation for sample preparation for elemental and chromatographic analysis. We have been deeply involved in the cannabis industry for many years and are familiar with testing procedures and regulations. We are founding members of many cannabis committees that are working on standardization of testing and reporting of data. We are passionate about EXTRACTION transcribing this information into systems to help your business grow and have the knowledge to support you along the way.

CHIEF SERVICES SUPPORTED • MARS 6: Microwave digestion system for World-class application and service support trace metals sample prep of all cannabis are what set CEM apart from other instrument and cannabis containing products. manufacturers. Our mission is to simplify your testing procedures with easy-to-use systems STATES SERVED that do the work for you. With built in meth- Nationwide ods, onboard support videos, and live applications and service support ready to help when you need it most, CEM is your partner in sample prep.

MAJOR PRODUCTS • EDGE: Automated extraction system for potency, pesticides, terpenes, and mycotoxins. The EDGE automatically performs solvent addition, sample extraction, fi ltration, and cooling in as little as 5 minutes.

CEM Corporation

3100 Smith Farm Road Matthews, NC 28104 USA

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LOCATIONS OF OTHER OFFICES AND FACILITIES France, Germany, Ireland, Italy, Japan, United Kingdom, Local agency representatives in all world regions www.cem.com

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Chem Service, Inc. COMPANY DESCRIPTION ANALYTICAL Established in 1962, Chem Service is accredited for ISO Guide 34:2009 and ISO/IEC 17025:2005 INSTRUMENTS and registered and certifi ed to the ISO 9001:2008 Quality Management System for the design, development, production, distribution, and servicing of organic neat and synthetic reference materials.

CHIEF SERVICES SUPPORTED • USP standards MANUFACTURING Chem Service, Inc., produces high purity • ISO standards & PROCESSING chemicals for use as reference materials and • DIN standards for other laboratory purposes. More than 95% • Explosive residues of their standards grade materials have a cer- • PCB congeners tifi ed purity of 98.0% or greater. Standards- • Petroleum hydrocarbons grade chemicals are clearly labeled with an • Fatty acid methyl esters OTHER expiration date that is based on years of expe- • High purity inorganics SERVICES rience in handling and testing. Products are • Surfactants packaged in small quantities to minimize stor- • Anti-oxidants age, waste, and disposal requirements. Or- • Ultraviolet inhibitors ganic and inorganic chemicals, solutions, and • Plasticizers mixtures are available to meet a wide range of • Biological stains specialized laboratory needs. • Phthalate standards • Polynuclear aromatics MAJOR PRODUCTS • Vitamins • Pesticide residue standards • Carbohydrates standards • Cannabis pesticide standards • Forensic standards • Residue solvent standards • Personal care product standards • Flame retardants/PBDE • Metabolite standards STATES SERVED • EPA standards US and international

Chem Service, Inc.

600 Tower Lane West Chester, PA 19380 USA

TELEPHONE Toll free: (800) 452-9994 (610) 692-3026 Fax: (610) 692-8729

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Frontier Laboratories Edibles quick and easy by EGA/PY 3030D Multi-Mode Pyrolyzer ANALYTICAL INSTRUMENTS COMPANY DESCRIPTION Frontier Laboratories, Ltd., was founded in 1991 by Dr. Chu Watanabe. His experience working at Hewlett-Packard’s Analytical Division and Dow Chemical made him uniquely qualifi ed to design and market analytical instruments for materials characterization. Dr. Watanabe, with the support of polymer scientists at Nagoya University in Japan, developed a pyrolyzer based on a vertical microfurnace design. Today the patented fourth generation vertical microfurnace serves as the corner- stone for the 3000 Series of products developed and marketed worldwide by Frontier Laboratories.

CHIEF SERVICES SUPPORTED STATES SERVED Research, development, and manufacture of Nationwide for the U.S. and Canada, as well analytical equipment. as Asia/Oceania, Europe, Middle East/Africa, and Russia. MAJOR PRODUCTS Frontier Laboratories, Ltd., designs and manufactures analytical instruments for materials characterization. The main products, supported by a number of accessories and software, include the EGA/PY-3030D Multi-function- al Pyrolysis System, the PY-3030S Single-Shot Pyrolyzer, the Rx-3050 series of Rapid Screen- ing Reactors for catalyst screening, and a line of Ultra ALLOY® stainless steel capillary columns. Frontier Labs’ products are compatible with most gas chromatographs and mass spectrometers from major manufacturers.

Frontier Laboratories

North American Technology Center: 5141 Lone Tree Way, Antioch, CA 94531

TELEPHONE (925) 813-04980

LOCATIONS OF OTHER OFFICES AND FACILITIES 4-16-20 Saikon Koriyama, Fukushima 963-8862, Japan

KEY PERSONNEL Rojin Belganeh, Technical Director- North America Itsuko Iwai, Senior Research Chemist- North America William Pipkin, Global Consulting www.frontier-lab.com

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Restek Corporation COMPANY DESCRIPTION ANALYTICAL For more than 30 years, Restek has been a leader in developing technologies and manufacturing INSTRUMENTS products for gas and liquid chromatography (GC and LC), including columns, reference standards, sample preparation materials, accessories, and more. We have decades of hands-on practical experience in chemistry, chromatography, and engineering, and our reputation for going the extra mile with our Plus 1 customer service and top-performing products is well known throughout the chromatography community. Restek has been providing the cannabis market with testing consumables and services since 2010 for both product safety and product quality. This includes CONSULTING supporting testing for potency, pesticides, residual solvents, mycotoxins, and terpenes.

CHIEF SERVICES SUPPORTED solvents, mycotoxins, and terpenes. Our Whether you are part of a well-established chemists develop LC and GC columns, ref- quality assurance lab or starting a new lab, erence standards, and sample preparation OTHER Restek has the products and expertise you products that streamline cannabis testing. SERVICES need for successful cannabis analysis. Being Restek’s Rxi columns deliver more accurate, an employee-owned and independent chro- reliable results than any other fused silica col- matography company, every employee at umn on the market. Our Raptor LC columns Restek has a vested interest in your success. are ideal for cannabis testing because they We design the best solutions for your lab, re- quickly separate your target compounds, pro- gardless of the instrumentation and tech- viding higher sample throughput. Restek is niques used. also continually expanding our product line • LC in order to meet the evolving needs of the • GC cannabis industry. Restek’s certifi ed refer- • Sample preparation ence materials are manufactured and quality • Reference standards control tested under ISO 17034 and ISO/EIC 17025 accreditations. MAJOR PRODUCTS We offer a range of chromatography con- STATES SERVED sumables for potency, pesticides, residual Nationwide and North America.

Restek Corporation

110 Benner Circle Bellefonte, PA 16823 USA

TELEPHONE Toll free: 1-800-353-1300

EMAIL [email protected]

KEY PERSONNEL Ashlee Gerardi, Business Development Manager - Cannabis Colton Myers, Applications Chemist – GC Solutions Justin Steimling, Applications Chemist – LC Solutions www.restek.com

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Shimadzu Scientifi c Instruments COMPANY DESCRIPTION ANALYTICAL Shimadzu is one of the world’s leading providers of analytical instruments for applications in a INSTRUMENTS broad range of industries. In the United States, Shimadzu has been at the forefront of working with cannabis testing laboratories and has a comprehensive understanding of the requirements of this evolving industry. From seed to sale, from accurate potency testing and terpene profi l- ing to contaminate testing for pesticides, residual solvents, heavy metals, and mycotoxins/afl a- toxins, Shimadzu offers scalable solutions to meet your testing needs for today and tomorrow.

CHIEF SERVICES SUPPORTED scientists is readily available to help your can- As medicinal and recreational cannabis mar- nabis testing laboratory succeed, assisting kets continue to grow, analytical testing will with method development, instrument train- ensure that consumers are receiving accurate- ing, and maintenance to ensure your systems ly labeled products that are free from contam- operate at an exceptional level. ination. Shimadzu is ready to assist you as you grow your laboratory. We offer a full suite of MAJOR PRODUCTS testing instruments, research platforms, sci- Shimadzu offers a full suite of analytical instru- entifi cally validated methods, and a varie- mentation to help grow your lab. These instru- ty of leasing programs to meet evolving re- ments include: quirements. In addition, our expert team of • HPLC • GC • GC–MS/MS • LC–MS/MS • AA • ICP-MS • Moisture balances • Online SFE-SFC-LC–MS/MS • MALDI-TOF MS • Consumables

STATES SERVED Nationwide

Shimadzu Scientific Instruments

7102 Riverwood Drive Columbia, MD 21046 USA

TELEPHONE Toll free: (800) 477-1227 (410) 381-1227 Fax: (410) 381-1222

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Pixis Labs COMPANY DESCRIPTION ANALYTICAL Pixis Labs sets the standard for quality analytics in the CBD and hemp markets nationwide and TESTING LAB raises the bar for Oregon cannabis laboratories. In partnership with Columbia Food Labs, and as a member of Tentamus Group, Pixis Labs boasts top-of-the-line instrumentation and nearly 40 years of analytical expertise. With low detection limits and robust methodologies, Pixis Labs is ahead of the curve for changing requirements and emerging markets, offering services beyond compliance. We pride ourselves on building a community of sustainable business partner- ships through detailed analyses and accurate reporting. Contact us for a custom service and price quote.

CHIEF SERVICES SUPPORTED MAJOR PRODUCTS Pixis Labs offers you reliable data and turn- Pixis Labs offers accurate and clear reporting, around times with transparent, professional catered to your market and industry. communication and competitive pricing. Consultation services are available upon • Potency HPLC (ISO 17025) request. • Pesticides by LC–MS (ISO 17025) • Heavy metals by MS/MS (ISO 17025) STATES SERVED • Residual Solvents by GC–MS (ISO 17025) Pixis Labs’ services are available nationally for • Organic-equivalent pesticide screen (ISO hemp and CBD analyses, as well as nutrition- 17025), developed in-house, by LC–MS al, environmental, and microbiological testing. • Mycotoxins by LC–MS/MS (ISO 17025) We are proud to serve the Oregon cannabis • Moisture, water activity market for compliance and R&D testing. • Terpenes by GC–MS • Pathogens • Yeast and mold • Nutritional lab: FDA Labels, allergens, vitamins, supplements, shelf life, and more

Pixis Labs

12423 NE Whitaker Way Portland, OR 97230 USA

TELEPHONE (503) 254 1794

EMAIL customerservice@ pixislabs.com

KEY PERSONNEL Derrick Tanner, General Manager Jen Stiles, Operations Manager Kelly O’Connor, Sales Rep www.columbiafoodlab.com

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SPEX CertiPrep COMPANY DESCRIPTION ANALYTICAL SPEX CertiPrep has been servicing the scientifi c community since 1954. We are a leading TESTING LAB manufacturer of certifi ed reference materials (CRMs) and calibration standards for analytical spectroscopy and chromatography. We offer a full range of inorganic and organic CRMs. We are certifi ed by DQS to ISO 9001:2015 and are proud to be accredited by A2LA to ISO/IEC 17025:2005 and ISO 17034:2016. The scope of our accreditation is the most comprehensive in the industry and encompasses all our manufactured products. ANALYTICAL INSTRUMENTS MAJOR PRODUCTS We offer analytical standards for medicinal and recreational cannabis testing. SPEX CertiPrep offers ISO 17034 CRMs for all of the common contaminants such as pesticide residues, residual solvents, and heavy metals EXTRACTION as well as qualitative analysis CRMs such as terpenes and cannabinoids. As the industry demands change and regulations are put into place, we continually update our product offerings. MANUFACTURING & PROCESSING STATES SERVED Nationwide for the US and Canada.

SPEX CertiPrep

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LOCATIONS OF OTHER OFFICES AND FACILITIES Stanmore, UK +44 (0) 208 204 6656

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VividGro COMPANY DESCRIPTION CULTIVATION/ VividGro is a pioneer in the AgTech space. Tailored to support the automation and effi ciency GROWING needs of the indoor agriculture and horticultural markets, VividGro implements solutions that help growers maximize yields and reduce costs. The state-of-the-art, customized lighting product line delivers optimized PAR to maximize plant growth and PAR effi cacy. Our control automation systems and data collection strategies are able to measure and modify growing to help growers learn how to use their resources more effi ciently. It is not the lights you use; it is how you MANUFACTURING use them. Learn more at VividGro.com. & PROCESSING CHIEF SERVICES SUPPORTED • FlowerMax (6 channel) • Custom LED grow lights, • FlowerMax Pro (8 channel) automation, and controls • GroNet (automation and control system)

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VividGro

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FRITSCH Milling & Sizing, Inc. COMPANY DESCRIPTION MANUFACTURING FRITSCH is an internationally respected German manufacturer of laboratory and produc- & PROCESSING tion-scale instruments used for milling, particle size analysis, and sample handling. Serving a broad range of industries, including food, pharmaceuticals, agriculture, and nano-technology, FRITSCH has worked closely with the medical cannabis industry to optimize a range of solutions for use in production, quality control, and R&D.

ANALYTICAL PRIMARY APPLICATIONS SUPPORTED REGIONS SERVED INSTRUMENTS • Production milling—optimize particle Global size to maximize extraction yield, homogenize solid extracts and ingredients, create Premium prerolls. • Quality control—homogenize edibles and plant samples to allow representative subsampling • Particle size analysis of milled materials

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FRITSCH Milling & Sizing, Inc.

57 Grant Drive, Suite G Pittsboro, NC 27312 USA

TELEPHONE (919) 229-0599

EMAIL [email protected]

LOCATIONS OF OTHER OFFICES AND FACILITIES Global Headquarters: Idar-Oberstein, Germany Company Offices: USA, France, Russia, Singapore, China Local agency representatives www.fritsch-us.com in all world regions www.cannabis-milling.com FOUNDING SPONSOR

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LGC Standards COMPANY DESCRIPTION MANUFACTURING LGC Standards is a global manufacturer and distributor of high quality reference materials with & PROCESSING a portfolio of more than 100,000 certifi ed reference materials, standards, and profi ciency testing schemes for the pharmaceutical (Mikromol™), environmental (Dr. Ehrenstorfer™), cannabis, food and beverage, forensic, clinical, industrial, and petrochemical markets. Our ISO accreditations include: ISO 9001, ISO 17034/ISO Guide 34, GMP/GLP, ISO 13485, ISO/IEC 17025, and ISO/ IEC 17043. As a designated National Measurement Institute (NMI) for chemical and bio-measurement, LGC works in partnership with governments, intermediaries, and private sector organizations to address the measurement needs of key industries including healthcare, food, environmental, security, and energy.

MAJOR PRODUCTS Dr. Ehrenstorfer™ offers a broad range of state-by-state cannabis impurity standards as well as a full list of terpenes to meet your state’s requirements. We manufacture standards for the analysis of pesticide, solvent, heavy metal, mycotoxin, bacteria, and growth regulator residues. All of our Dr. Ehrenstor- fer™ brand mixes are certifi ed reference materials manufactured in accordance with ISO 17034 requirements. Custom mixes are our specialty.

STATES SERVED Global

LGC Standards

276 Abby Road Manchester, NH 03103 USA

TELEPHONE Toll free: (888) 622-7660 (603) 622-7660 Fax: (603) 622-5180

EMAIL [email protected]

KEY PERSONNEL Don Shelly, Food and Environmental Product Manager Adam Ross, Environmental, Cannabis, Food & Beverage Specialist https://us.lgcstandards.com

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PRO Scientifi c Inc. COMPANY DESCRIPTION MANUFACTURING PRO Scientifi c is a global leader in the manufacturing of homogenizers for both laboratory and AND PROCESSING industrial labs requiring sample preparation. PRO homogenizers are able to homogenize various forms of cannabis material fast and effi ciently. From micro sample volumes to larger multi-liter processing, there is a PRO Homogenizer to suit your needs. PRO Scientifi c even offers homogenizing solutions to address automated multi-sample homogenizing and OEM homogenizer needs too. All products manufactured by PRO ANALYTICAL Scientifi c are backed with over 25 years of technical experience, unmatched customer support, INSTRUMENTS and made in the USA.

CHIEF SERVICES SUPPORTED reliable, reproducible results that are required PRO Scientifi c is the ideal source for high for this industry. A range of setups are avail- quality homogenizing products for sample able depending upon your budget and pro- OTHER preparation within the cannabis industry, processing requirements. Whether high speed SERVICES viding assistance for both small startup and shearing to dispersing, homogenizing, emul- established labs with their cannabis testing sifying, mixing, or blending. and quality control. PRO Scientifi c homoge- • Economical homogenizing package kits nizers are precision homogenizers to assist in • Handheld or stand mounted determining and monitoring the cannabinoid homogenizer models potency and pesticide residue. PRO Scientif- • Digital and programmable ic Homogenizers are also ideal for the extrac- benchtop homogenizer models tion of cannabis for the development of can- • Automated and semi-automated nabis infused products. Homogenization of homogenizing systems cannabis samples allows for a stable emulsion • Precision crafted generator probes to be created that is shelf stable. STATES SERVED MAJOR PRODUCTS PRO Scientifi c supports labs in various indus- Our PRO Homogenizers and generator tries that require sample preparation through- probes are precision designed to provide out the United States as well as internationally.

PRO Scientific Inc.

99 Willenbrock Road Oxford, CT 06478 USA

TELEPHONE Toll free: (800) 584-3776 (203) 267-4600 Fax: (203) 267-4606

EMAIL [email protected]

KEY PERSONNEL Holly Yacko-Archibald, Vice President Brian Archibald, Business Development Manager www.proscientific.com

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Vitalis Extraction Technology COMPANY DESCRIPTION industry. Vitalis’ award-winning systems focus MANUFACTURING Vitalis Extraction Technology Inc.© (Vitalis) is a on the key needs of any large-scale extrac- & PROCESSING privately-owned, Kelowna-based engineering tion business: reliability, component certifi - and manufacturing company, producing the cation, and continuous 24/7 operation. Vitalis

highest-fl owing industrial supercritical CO2 systems were engineered to balance all three extraction systems for the cannabis industry. fundamentals, and the result is the highest Renowned for their reliability, scalability, and output of the purest full-spectrum oil possible

continuous operation, Vitalis systems are de- in the shortest amount of time in the CO2 ex- EXTRACTION signed and manufactured in accordance with traction industry. ASME and CSA Standards for Boiler, Pressure Vitalis produces three lines of CO2 extrac- Vessel and Pressure Piping Code. The ves- tion equipment: The F-Series, the Q-Series, sels are stamped with a CRN and NB registra- and the R-Series. The R-Series’ R-400 allows tion number confi rming that the vessels meet for processing of up to 500 pounds of top- code and have been examined by an Author- quality material per day. With a heavy-duty ized Inspector. With systems on three conti- hydraulic pump, dual extraction lines, indus-

nents, Vitalis has the most deployments of in- trial grade CO2 recovery system, TrueCyclon-

dustrial CO2 supercritical extractors into the ic separation technology, and modular assem- cannabis industry. bly for future scalability, the Vitalis system is built for heavy-duty workloads, massive ca- MAJOR PRODUCTS pacity, and continual growth. With a strict Vitalis Extraction Technology is the leading focus on quality as well as a commitment to manufacturer of the most sophisticated in- constant research and development, Vital-

dustrial Supercritical CO2 extraction systems is provides certifi ed Supercritical CO2 extrac- in the world. Vitalis is Canada’s fi rst and only tion equipment that is redefi ning the capabil- Vitalis Extraction American Society of Mechanical Engineers ities of traditional extraction methodologies. Technology (ASME) certifi ed extraction original equip- #102-480 Neave Court ment manufacturer (OEM) in the cannabis STATES SERVED Kelowna, BC Global—systems deployed across North V1V 2M2 Canada America, South America, and Europe.

TELEPHONE Toll free: (844) 248-2326 (250) 864-4015

EMAIL [email protected]

LOCATIONS OF OTHER OFFICES AND FACILITIES Sales Office 3314 Appaloossa Road Kelowna, BC V1V 2W5

Operations & Assembly 591 Gaston Avenue Kelowna, BC V1Y 7E6

KEY PERSONNEL Pete Patterson, COO, Cofounder Joel Sherlock, Chairman, Cofounder James Seabrook, CTO, Cofounder https://vitaliset.com/

CANNABIS SCIENCE AND TECHNOLOGY | www.CannabisScienceTech.com VOL 1 • NO 4 • NOVEMBER/DECEMBER 2018 Supplier Proﬁ le 77

COMPLED Solutions GmbH COMPANY DESCRIPTION OTHER COMPLED was founded in 2010 and has become one of the pioneers in LED horticulture light- SERVICES ing. Since then COMPLED has been in the forefront of LED horticulture lighting for science applications. We develop, manufacture, and test exclusively in Germany. We have always been focused on tunable light spectra because that’s the mayor advantage of LED technology. By dy- namically tuning the spectrum it is possible to optimize yields as well as cannabinoid and terpene profi les. Furthermore, it is also crucial to align lighting recipes to all other relevant envi-

ronmental parameters such as CO2, temperature, and so forth, and that’s actually also possible with our technology.

CHIEF SERVICES SUPPORTED impact of the quantity and quality of light on Besides horticulture lighting design we also the development of plants. We offer several offer support in the development of lighting solutions from 24 LED channels down to six and growing recipes, data science, and bio- LED channels. logical research related to lighting. Even cus- In addition, we are proud to offer our fi rst tomized lighting devices or spectra are avail- LED horticulture lighting platform especially able upon request. Please have a look at our designed for cannabis cultivation, the Hash- references (www.compled.de/pages/refer- Cropter platform (www.hashcropter.com). It’s ences.html). the most fl exible and effective LED lighting platform for cannabis cultivation. MAJOR PRODUCTS Spectral tuning is always obligatory and Our main product line is called the SUNsim both platforms are combinable. platform (www.compled.de/pages/products. html#research_jump). It was designed espe- STATES SERVED cially for researchers who are interested in the Nationwide and worldwide.

COMPLED Solutions GmbH

Dresden, Saxony, D-01324 Germany

TELEPHONE +49-351-873234-00 Fax: +49-351-873234-01

EMAIL [email protected]

KEY PERSONNEL Christoph Schubert, CEO www.compled.de

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LEAP PAL Parts +Consumables COMPANY DESCRIPTION OTHER LEAP PAL Parts + Consumables has been helping customers in the lab instrumentation and liq- SERVICES uid handling business for more than 25 years. We represent the highest quality products that meet strict quality assurance (QA) standards and work with the best manufacturers to offer innovative products at the most competitive prices. We provide same day shipping of all in-stock products to avoid instrument downtime as well as reduce your on-hand inventory levels. Our mission statement is: Be Good. Just that simple. Be good to our customers, our suppliers, our ANALYTICAL community, and to each other. Let us show you what thousands of customers already know. INSTRUMENTS CHIEF SERVICES SUPPORTED include CTC Analytics, VALCO Instruments, We offer a complete range of consumables IDEX/Rheodyne, La-Pha-Pack and many for high pressure liquid chromatography, gas others. chromatography, headspace, and sample ANALYTICAL preparation. In addition to consumables, we STATES SERVED TESTING LAB offer instrument specifi c small parts. As an We serve all 50 states and Canada. Providing added service we also provide technical assis- products internationally and domestically, to tance by certifi ed PAL, PAL-XT, PAL 3 service customers in the following industries: technicians if you need urgent assistance with • Biotechnology your instruments. • Pharmaceutical MANUFACTURING • Clinical & PROCESSING MAJOR PRODUCTS • Food and beverage We provide syringes and syringe compo- • Environmental nents, vials, caps and inserts including our • Petrochemical own L-MARK brand, instrument parts and ac- • Cannabis analysis cessories, valve components including ro- • Forensics PACKAGING & tors, valves, tubing, fi ttings and sample loops • Toxicology SUPPLIES as well as plates and cap mats. Our suppliers

LEAP PAL Parts +Consumables

4216 Atlantic Ave. Raleigh, NC 27604 USA

TELEPHONE Toll free: 1-877-725-1007 (919) 322-4208 Fax: (888) 589-9475

EMAIL [email protected]

KEY PERSONNEL Lamar Jones, President Kate Holub, VP of Sales Angie Buchholz, Marketing Director www.palparts.com

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