Potency
Solvent
Microbial
Terpene
At this time the Alaska’s Marijuana Control Board requires concentration testing of the first five cannabinoids listed below. CannTest, LLC will include the additional significant cannabinoids in the profile, and add new compounds as science evolves.
At this time the Alaska’s Marijuana Control Board requires concentration testing of the first five cannabinoids listed below. CannTest, LLC will include the additional significant cannabinoids in the profile, and add new compounds as science evolves.
There are presently two methods used for potency testing of cannabinoids (THC, THCA, CBD, CBDA, CBN and others). The method of Gas Chromatography (GC) requires that a cannabis sample first be ground to a fine powder. Addition of an organic solvent such as methanol followed by sonification and centrifuging is then used to extract cannabinoids from the plant material. A small amount of the sample is injected into a GC machine where it is heated up to 300 degrees Celsius thereby vaporizing the cannabinoids into a gaseous mixture. The gas is fed through a column that through its chemical makeup retains some compounds longer than others. As each cannabinoid leaves the column at a unique time a flame burns the gas and a detector determines the amount of each cannabinoid passing through the column. This is termed Flame Ionization Detection (FID).
The problem with Gas Chromatography is that much of the natural cannabinoids in cannabis exist in an acid form such as THC-A and CBD-A then decarboxylate (release carbon dioxide) to the neutral forms of THC and CBD. THC has a psychoactive effect, but THC-A does not. Typically when cannabis is smoked or vaporized THC-A transforms to THC resulting in the “high”. However research has indicated that MS patients have had a therapeutic effect with intake of unheated cannabis due to anti-inflammatory effects of THC-A. Recent lab studies have shown that a GC machine does not decarboxlyate 100% of the THC-A and CBD-A. Roughly 60-70% was the average rate.
The second method of testing of cannabinoid potency is High Performance Liquid Chromatography (HPLC). In the HPLC method a cannabis sample is ground to a fine powder and cannabinoids are extracted using a solvent, sonification and centrifuging as in GC. However in HPLC the sample passes through a column in a liquid phase and cannabinoids are detected by an untraviolet light. Because HPLC does not add heat to the system it allows for the unique determination of THC-A and CBD-A as well as THC and CBD.
CannTest, LLC will use the LC method of testing for cannabinoid potency as it it the most accurate means of determining the Total THC and CBD that will be provided on delivery. Less than 100% decarboxylation during GC means that the total amount of THC and CBD can not be known. The GC system will only measure the THC available at the detector. In addition GC can not produce a measurement of THC-A and CBD-A which will be valuable information for many medical users.
There are presently two methods used for potency testing of cannabinoids (THC, THCA, CBD, CBDA, CBN and others). The method of Gas Chromatography (GC) requires that a cannabis sample first be ground to a fine powder. Addition of an organic solvent such as methanol followed by sonification and centrifuging is then used to extract cannabinoids from the plant material. A small amount of the sample is injected into a GC machine where it is heated up to 300 degrees Celsius thereby vaporizing the cannabinoids into a gaseous mixture. The gas is fed through a column that through its chemical makeup retains some compounds longer than others. As each cannabinoid leaves the column at a unique time a flame burns the gas and a detector determines the amount of each cannabinoid passing through the column. This is termed Flame Ionization Detection (FID).
The problem with Gas Chromatography is that much of the natural cannabinoids in cannabis exist in an acid form such as THC-A and CBD-A then decarboxylate (release carbon dioxide) to the neutral forms of THC and CBD. THC has a psychoactive effect, but THC-A does not. Typically when cannabis is smoked or vaporized THC-A transforms to THC resulting in the “high”. However research has indicated that MS patients have had a therapeutic effect with intake of unheated cannabis due to anti-inflammatory effects of THC-A. Recent lab studies have shown that a GC machine does not decarboxlyate 100% of the THC-A and CBD-A. Roughly 60-70% was the average rate.
The second method of testing of cannabinoid potency is High Performance Liquid Chromatography (HPLC). In the HPLC method a cannabis sample is ground to a fine powder and cannabinoids are extracted using a solvent, sonification and centrifuging as in GC. However in HPLC the sample passes through a column in a liquid phase and cannabinoids are detected by an untraviolet light. Because HPLC does not add heat to the system it allows for the unique determination of THC-A and CBD-A as well as THC and CBD.
CannTest, LLC will use the LC method of testing for cannabinoid potency as it it the most accurate means of determining the Total THC and CBD that will be provided on delivery. Less than 100% decarboxylation during GC means that the total amount of THC and CBD can not be known. The GC system will only measure the THC available at the detector. In addition GC can not produce a measurement of THC-A and CBD-A which will be valuable information for many medical users.
Current regulations allow for THC potency up to 76 percent. To achieve these high levels of THC marijuana production facilities produce solvent-based marijuana concentrate by using the hydrocarbons N-butane, isobutane, propane, or heptane to extract the cannabinoid from plant material. After extraction is complete the solvent must be purged from the product through evaporation. Often there is not complete evaporation of solvents leaving dangerous chemical in the concentrate.
The following solvents will be tested for in all marijuana concentrates. Values below are given in Parts Per Million (PPM). 1% is equivalent to 10,000PPM. Therefore if there is a 1 gram sample of concentrate the upper limit of butane allowed would be 800/10,000 x 0.01 or 0.08mg. Those with limits higher than the values below will be rejected and not sold in retail outlets.
Current regulations allow for THC potency up to 76 percent. To achieve these high levels of THC marijuana production facilities produce solvent-based marijuana concentrate by using the hydrocarbons N-butane, isobutane, propane, or heptane to extract the cannabinoid from plant material. After extraction is complete the solvent must be purged from the product through evaporation. Often there is not complete evaporation of solvents leaving dangerous chemical in the concentrate.
The following solvents will be tested for in all marijuana concentrates. Values below are given in Parts Per Million (PPM). 1% is equivalent to 10,000PPM. Therefore if there is a 1 gram sample of concentrate the upper limit of butane allowed would be 800/10,000 x 0.01 or 0.08mg. Those with limits higher than the values below will be rejected and not sold in retail outlets.
Residual Solvent testing uses Gas Chromatography (GC) with Flame Ionization Detection (FID) and Headspace Injection. To conduct this test first a very small sample (50mg) is heated to 140 degrees Celsius for 30 minutes in a sealed environment (the Headspace). This creates a gaseous mixture of residual solvents. The sample gas is fed into a Gas Chromatography (GC) column through the use of a carrier gas such as Helium. The GC column, due to the chemical properties of the column liner, reacts uniquely with each gas present in the mixture.
Each compound takes a specific amount of time to pass through the column. As the compounds exit the column they are detected by the GC and sent to a computer processing system. The program produces a graph or chromatogram of residual solvents elicited from the column over a period of time. For the peaks of the chromatogram to have quantitative meaning a calibration must be done prior to testing. A residual solvent mixture of known concentration is procured from a company such as Restek, Inc. The sample is diluted to make mixtures of several different concentrations. Each concentration is tested as per the above description to create a calibration curve. The peaks of the sample can than be compared to the calibration curve to determine the actual concentration of each compound in the chromatogram.
Residual Solvent testing uses Gas Chromatography (GC) with Flame Ionization Detection (FID) and Headspace Injection. To conduct this test first a very small sample (50mg) is heated to 140 degrees Celsius for 30 minutes in a sealed environment (the Headspace). This creates a gaseous mixture of residual solvents. The sample gas is fed into a Gas Chromatography (GC) column through the use of a carrier gas such as Helium. The GC column, due to the chemical properties of the column liner, reacts uniquely with each gas present in the mixture.
Each compound takes a specific amount of time to pass through the column. As the compounds exit the column they are detected by the GC and sent to a computer processing system. The program produces a graph or chromatogram of residual solvents elicited from the column over a period of time. For the peaks of the chromatogram to have quantitative meaning a calibration must be done prior to testing. A residual solvent mixture of known concentration is procured from a company such as Restek, Inc. The sample is diluted to make mixtures of several different concentrations. Each concentration is tested as per the above description to create a calibration curve. The peaks of the sample can than be compared to the calibration curve to determine the actual concentration of each compound in the chromatogram.
Cannabis that is grown outside ideal temperature and humidity conditions can develop mold during growth. Improper conditions during the drying process can also result in mold growth. Common molds found in improperly handled cannabis include Cladosporium, Penicillium, Alternaria, Aspergillus and Mucur. Bacteria associated with cannabis include E.coli, Salmonella and Listeria. Molds produce toxins that can result in serious health concerns including cancer, bronchpulmonary complications, sinusitis and gastrointestinal problems. CannTest, LLC will use a process developed by the U.S. Pharmacopeial Convention and detailed in their US 2021 and USP 61 documents. The U.S. Pharmacopeial Convention (USP) is a scientific nonprofit organization that since 1820 has set standards for the identity, strength, quality, and purity of medicines, food ingredients, and dietary supplements manufactured, distributed and consumed worldwide. Testing utilizing this process will allow ensure that cannabis that has been produced without using sound agricultural practices to prevent the development of mold and bacteria will not make it to the shelves of Alaska’s dispensaries.
The Marijuana Control Board currently requires testing for the following bacteria and fungi.
CannTest LLC will use quantitative real-time polymerase chain reaction (qPCR) to conduct testing for bacteria and molds. qPCR is a relatively new method which provides significant advantages over plate culture techniques which had previously been the standard for microbiological testing. Plate culture techniques take up to seven days to produce results, and those results are subject to human interpretation. qPCR produces results in few hours through using a thermostable enzyme to double the quantity of a short specific part of the target microbe (E-Coli, Salmonella, etc.) DNA in successive heating/cooling cycles. In every cycle the number of short specific sections of DNA is doubled, leading to an exponential amplification of targets. The amplified DNA is fluorescently labeled and the amount of the fluorescence released during amplification can be measured in direct proportion to the amount of amplified DNA. The higher the initial number of DNA molecules in the sample, the faster the fluorescence will increase during the successive qPCR cycles. In other words, if a sample contains more DNA targets, the fluorescence will be detected in earlier cycles.
The number of cycles required to produce a given amount of flourescence is termed quantitation cycle (Ct for short) and is the basic data output unit of qPCR. The lower Ct values mean higher initial copy numbers of the target DNA. CannTest, LLC will use this fast and accurate method of qPCR to perform all required microbiological testing.
CannTest LLC will use quantitative real-time polymerase chain reaction (qPCR) to conduct testing for bacteria and molds. qPCR is a relatively new method which provides significant advantages over plate culture techniques which had previously been the standard for microbiological testing. Plate culture techniques take up to seven days to produce results, and those results are subject to human interpretation. qPCR produces results in few hours through using a thermostable enzyme to double the quantity of a short specific part of the target microbe (E-Coli, Salmonella, etc.) DNA in successive heating/cooling cycles. In every cycle the number of short specific sections of DNA is doubled, leading to an exponential amplification of targets. The amplified DNA is fluorescently labeled and the amount of the fluorescence released during amplification can be measured in direct proportion to the amount of amplified DNA. The higher the initial number of DNA molecules in the sample, the faster the fluorescence will increase during the successive qPCR cycles. In other words, if a sample contains more DNA targets, the fluorescence will be detected in earlier cycles.
The number of cycles required to produce a given amount of flourescence is termed quantitation cycle (Ct for short) and is the basic data output unit of qPCR. The lower Ct values mean higher initial copy numbers of the target DNA. CannTest, LLC will use this fast and accurate method of qPCR to perform all required microbiological testing.
Terpenes are found in a wide variety of plants. They are responsible for the aroma and flavor of the plant. In cannabis they are also responsible for modulating the effect of cannabinoids on receptors within the body. Researchers have dubbed this the “Entourage Effect”. There is still much research that needs to be done to clearly identify the methods of synergistic interaction of these compounds, but for an introduction to this subject please visit this link to a paper in the British Journal of Pharmacology. Research studies have identified the effects of individual terpenes on human physiology.
Although testing for terpenes is not currently required by the Marijuana Control Board, CannTest, LLC will offer a test for concentrations of prominent cannabis terpenes listed below. The stand alone effect and where, other than cannabis, the substance is commonly found are also listed. CannTest, LLC will closely monitor research into the “Entourage Effect” to identify additional impacts on the human system so that consumers can maximize understanding of their purchases.
Terpene testing uses Gas Chromatography (GC) with Flame Ionization Detection (FID) and Headspace Injection. To conduct this test first a very small sample (50mg) is heated to 140 degrees Celsius for 30 minutes in a sealed environment (the Headspace). This creates a gaseous mixture of terpenes. The sample gas is fed into a Gas Chromatography (GC) column through the use of a carrier gas such as Helium. The GC column, due to the chemical properties of the column liner, reacts uniquely with each gas present in the mixture.
Each compound takes a specific amount of time to pass through the column. As the compounds exit the column they are detected by the GC and sent to a computer processing system. The program produces a graph or chromatogram of terpenes elicited from the column over a period of time. For the peaks of the chromatogram to have quantitative meaning a calibration must be done prior to testing. A terpene mixture of known concentration is procured from a company such as Restek, Inc. The sample is diluted to make mixtures of several different concentrations. Each concentration is tested as per the above description to create a calibration curve. The peaks of the sample can than be compared to the calibration curve to determine the actual concentration of each compound in the chromatogram.
Terpene testing uses Gas Chromatography (GC) with Flame Ionization Detection (FID) and Headspace Injection. To conduct this test first a very small sample (50mg) is heated to 140 degrees Celsius for 30 minutes in a sealed environment (the Headspace). This creates a gaseous mixture of terpenes. The sample gas is fed into a Gas Chromatography (GC) column through the use of a carrier gas such as Helium. The GC column, due to the chemical properties of the column liner, reacts uniquely with each gas present in the mixture.
Each compound takes a specific amount of time to pass through the column. As the compounds exit the column they are detected by the GC and sent to a computer processing system. The program produces a graph or chromatogram of terpenes elicited from the column over a period of time. For the peaks of the chromatogram to have quantitative meaning a calibration must be done prior to testing. A terpene mixture of known concentration is procured from a company such as Restek, Inc. The sample is diluted to make mixtures of several different concentrations. Each concentration is tested as per the above description to create a calibration curve. The peaks of the sample can than be compared to the calibration curve to determine the actual concentration of each compound in the chromatogram.
