Quality of Analytical Testing. The quality of any results from analytical testing are inherently tied to several factors:
- Competence level and training records for staff
- Procedural compliance for all instrument calibration and analytical testing
The best way to ensure the above criteria are met is for any testing performed to be done so under Good Laboratory and Manufacturing practices. It requires the strict adherence to policies rich in documentation and record keeping
ARA – Avanti Rx Analytics uses the latest technology and established compendia and in-house validated methods to provide a full spectrum of Quality Assurance/ Quality Control analysis for cannabis and hemp, consisting of; (I) quantitative cannabinoid analysis (i.e., potency and profiling), (II) analysis of the essential oils to provide an aromatic profile (i.e., terpenes), (III) analysis of pesticide residue, (IV) analysis of possible contamination (i.e., heavy metals/trace element), (V) analysis of microbial contamination with fungi and bacteria, and (VI) determination of residual solvents
Morphological characteristics and variation in color of cannabis plants are influenced by the seed strain as well as by environmental factors such as light, water, nutrients and space
Dried and milled Cannabis sativa spp. is dark green to light green and tan colored flowering heads plant particulates with characteristic aroma having citrus and pine overtones.
Visual inspection should confirm the absence of absence of stalks, extraneous substances, insects and other vermin.
There is no requirement to mill or irradiate the dried marihuana, although the licensed producers may choose to do so.
Beginning with dry-weight determination, we analyze the percentage of water contained in each sample to confirm proper preparation during the curing process.
At ARA we utilize fully calibrated Mettler Toledo halogen drying moisture balances for determining moisture content in lab cannabis samples to obtain accurate and reliable moisture determination results to the fourth decimal point.
Moisture Analysis is also required for Cannabinoid potency determination and profiling of cannabis and hemp samples.
Cannabinoids occur naturally in significant quantity in the cannabis plant, and are concentrated in a viscous resin that is produced in glandular structures known as “Trichomes”.
The healthcare industry could not exist without a method to accurately dose medicine. In the case of the Cannabis plant there active ingredients are known as Cannabinoids. There are approximately 86 identified cannabinoids. Of these Cannabinoids the most known active in the medical research are Δ9-tetrahydrocannabinol (Δ9-THC); Cannabidiol(CBD); acid forms of Δ9-THC (Δ9-THCA) and CBD (CBDA), Cannabichromene (CBC); Cannabigerol (CBG); Δ8 - tetrahydrocannabinol (Δ8-THC); and the degradated resultant of Δ9-THC, cannabinol (CBN).
Cannabinoid profiling (potency) testing via high pressure liquid chromatography is highly recommended for products that will be ingested in a raw or unheated form, infused products, and for concentrate producers looking to test the completeness of their decarboxylation process. This method uses solvents rather than heat during the analysis, allowing for detection of non-psychoactive, acidic Cannabinoids like Δ9-THCA, CBDA, and CBGA, which form Δ9-THC and CBD when heated.
At ARA we accurately perform potency determination/quantification and profiling of 9 cannabinoids; THCV, CBD, CBG, CBDA, CBN, Δ9-THC, Δ8-THC, CBC and Δ9-THCA using in-house validated HPLC methods.
Tetrahydrocannabinol (THC) or more precisely its main isomer (−)-trans-Δ9-tetrahydrocannabinol, is the principal psychoactive constituent (or cannabinoid) of cannabis. It can be an amber or gold colored glassy solid when cold, which becomes viscous and sticky if warmed.
Δ8-Tetrahydrocannabinol (Δ8-THC) is an analog of the natural psychoactive compound Δ9-THC.
Tetrahydrocannabinolic Acid (THCA)
Tetrahydrocannabinolic acid (THCA, Δ9-THCA, 2-COOH-THC), is a biosynthetic precursor of tetrahydrocannabinol (THC), the active component of cannabis. When purified, it forms a powder which is unstable in the presence of acids, heat, oxygen, and/or light.
Cannabidiol is one of at least 113 active cannabinoids identified in cannabis. It is a major phytocannabinoid, accounting for up to 40% of the plant's extract
Cannabidiolic Acid (CBDA)
Cannabidiolic Acid (CBDa) is a cannabinoid found in the Cannabis plant.
Tetrahydrocannabivarin is a homologue of tetrahydrocannabinol having a propyl side chain instead of a pentyl group on the molecule, which makes it produce very different effects from THC
Cannabidiol (CBD) is one of at least 113 active cannabinoids identified in cannabis. It is a major phytocannabinoid, accounting for up to 40% of the plant's extract.
Cannabinol (CBN) is a weak psychoactive cannabinoid found only in trace amounts in Cannabis sativa and Cannabis indica. It is mostly a metabolite of tetrahydrocannabinol (THC).
Cannabichromene (CBC) is a cannabinoid found in the Cannabis plant. It bears structural similarity to the other natural cannabinoids
Aflatoxins, a mold largely produced by Aspergillus flavus and Aspergillus parasiticus are commonly tested mycotoxins found naturally in a wide range of agriculture crops including cannabis. Due to their harmful effects on human health, animal health, and global trade, aflatoxins are regulated in most countries and have established global limits in a wide variety of matrices. Regulations for the maximum limits vary for the reported aflatoxin B1 and total aflatoxins (sum of B1, B2, G1, and G2); however, the LPs must perform testing to approve the safety of their cannabis. Testing may often reveal aflatoxin levels above the maximum limits allowed, creating a trade restriction for certain agriculture and food products from certain countries3. The tests are performed according to their sampling methods and the results are measured against their established limits.
At ARA we use validated modified EP method for detecting and quantifying aflatoxins.
Medical marihuana producers, patients, and doctors are encouraged to well consider the dangers that Organic contaminants in marihuana born from microorganisms present.
Patients with serious health conditions, such as immune disorders cannot afford to be breathing in mold spores and nasty bio-toxins. As well, smoking moldy cannabis introduces spores into the lungs where they grow into large mold balls causing infection that is difficult to remedy and sometimes surgery is the only solution.
Under MMPR, it is the Must requirement that Medical Marihuana to be screened for mold, certain bacteria, mycotoxins (fungus metabolites), and fungus. The microbial and chemical contaminants of dried marihuana Must be within generally accepted tolerance limits for herbal medicines for human consumption, as established in any publication referred to in Schedule B of the Food and Drugs Act
European Pharmacopeia (EU.Phr.) chapter 5.8.1 (current edition): "Microbial Quality of Herbal Medicinal Products for Oral Use". In this case, testing for total aerobic microbial count (TAC), total combined yeast and molds count, bile-tolerant gram negative bacteria, Escherichia coli, and Salmonella Spores of bacteria and fungi are omnipresent in our biosphere and as a result may grow anywhere there is water, suitable temperature, and some sort of nutrition source. The presence of bacterial or fungal spores on a raw material does not suggest growth of that bacterium or fungus in any way. However, the abundance of these spores may indicate a general state of cleanliness that requires further testing to validate.
Gross mold infestation is often easy to spot. Trichomes are the crystal resin factories (biosynthesize THC, Terpenes).
The resin can be confused with mold. Mold grows on top of the trichomes and looks furry cotton candy under a magnifying glass. Trichomes look nothing like cotton candy; they are "clean" looking kind of like a glass fiber optic material.
Molds are ubiquitous, and small amounts are found in almost every cannabis sample. However, exposure to high levels of such microorganisms are known to cause health problems and can be particularly dangerous to patients that have existing medical conditions.
Mold concerns in marihuana are almost always due to moisture lock. If marihuana is packaged or cured in air-tight containers while buds are moist, this creates the ideal environment to incubate, or cultivate mold and microbes.
Mycotoxicoses are diseases caused by mycotoxins, i.e. secondary metabolites of molds. Both animal and human studies have found that in extreme cases, Mycotoxins can cause death. Exposure to mycotoxins is mostly by ingestion, but also occurs transdermally or by inhalation.
Aspergillus is a common, but toxic fungus that inhabits some marihuana plants, given the right conditions. Aspergillosis is a condition where the actual fungus infests the victim's body, particularly the lungs. Fungus and baby spores become concentrated in dark, damp and warm places. Susceptible individuals will experience allergic reaction, asthma and worse yet, they can get Aspergillosis
ARA – Avanti Rx Analytics offers microbiological method qualification and microbial content and bioburden screening of cannabis and hemp according to methods listed in the United States Pharmacopeia (USP), European Pharmacopoeia (Ph. Eur. or EP) under complete GMP quality systems and in compliance with Health Canada’s MMPR requirements.
Our microbial testing services include:
Requirements for sample preparation:
Adherence to Good Production Practices:
Section 52(1) of the MMPR prohibits a licensed producer from selling or providing dried marihuana unless the Good Production Practices set forth in Division 4 of the MMPR have been met. This includes the requirements for use of pesticides. The initial MMPR s54. (1) act published in June 19, 2013 stated marihuana must not be treated; before, during or after the drying process - with a pest control product that has not been registered under the Pest Control Products Act for use on marihuana for medical purposes. Under the Pest Control Products Act, pesticides may only be used if assessed and registered by the Pest Management Regulatory Agency (PMRA) for specific uses.
To date there are eleven registered pesticides approved by the Pest Management Regulatory Agency (PMRA) for use on cannabis (marihuana) that is produced commercially indoors. They are:
“Concerning testing of the above pesticides (i.e., insecticides, herbicides, and fungicides), this is required when a maximum residue limit is establish by PMRA. At the moment, none of these pesticides have a maximum residue limit. As a result, no testing is required as per the Pest Control Products Act.”
To date no specification nor analytical method/s for the above pesticides (i.e., insecticides, herbicides and fungicides) has been provided by both PMRA and Health Canada MMPR regulation. As well there are no direct instruction has been given on how and when to use these pesticide during cannabis production processes. The present regulation is quite vague when it comes to determination of recommended approved pesticides by the Pest Management Regulatory Agency (PMRA) for use on cannabis (marihuana).
Chemical pesticides are, by design are highly toxic and persistent. Many kill on contact and remain active for weeks, months, and even years. Human exposure to pesticides may lead to acute health problems, such as abdominal pain, dizziness, headaches, nausea, vomiting, as well as skin and eye problems. Many serious diseases, such as cancer, reproductive dysfunction, and asthma have been linked to pesticide exposure. While many growers and collective dispensaries take care to provide safe, effective products, still there are numerous reports of the presence of pesticides in purchased cannabis
Until clear direction and established methods are provided by the Health Canada, here at ARA – Avanti Rx Analytics will not be able to provide pesticide residue determination for the recommended pesticides shown in the above table.
ARA offer the following pesticide residue screening for the below pesticide however, these pesticides are not permitted under ACMPR (repealed MMPR) regulation and Must not be used for on cannabis (marihuana) that is produced commercially in Canada:
Cannabis concentrates can physically be produced in many different ways and some of these production techniques may use chemical solvents. The use of chemical solvents for the production of concentrates, to our understanding, is not yet been approved or clearly defined under Access to Cannabis for Medical Purposes Regulations (ACMPR) (i.e., repealed MMPR).
The presence of chemical residues within concentrated cannabis could have a significant negative impact on the health of those consuming the product. Dietary supplements, and other botanical products, provided to the general public are required to demonstrate the absence of residual solvents if any solvent was used in their production process. Medical cannabis concentrate producers should seek to establish their products are free and clean of solvents as well.
It is impossible to detect any residues remain in a concentrate when it was produced using solvents by smelling or visual inspection.
In order to address this matter with the appropriate sensitivity required, here at ARA – Avanti Rx Analytics, we have developed a fully validated method for the detection of a broad range of solvents that may be used in the production of cannabis concentrate.
Here at ARA we use validated methods utilizing a combination of head-space sampling, gas-chromatography with Flame Ionization Detector (GC/FID) or mass-spectrometry (GC/MS) that can detect, identify and quantify trace amounts of volatile solvents that might have been used in the production of cannabis concentrates:
An example of some of the solvents that can be detected:
Not only can we confirm if a solvent has been used, but we will also be able to determine which solvent was used. Our method is non-selective and can detect extremely trace amounts of any solvent that might have been used in cannabis concentrates production.
The cannabis plant consists of a wide variety of chemicals and compounds. About 140 of these belong to a large class of aromatic organic hydrocarbons known as Terpenes and Terpenoid the terms that are increasingly used interchangeably however, these terms do have different meanings. The main difference between terpenes and terpenoids is that terpenes are hydrocarbons (meaning the only elements present are carbon and hydrogen); whereas, terpenoids have been denatured by oxidation (drying and curing the flowers) or chemically modified. About 10-29 percent of marihuana smoke resin is composed of terpenes and terpenoids. Terpenes are ultimately responsible for delivering the complex effects provided by whole plant medicinal cannabis products. Cannabis age, maturation, and time of day can affect the amount and ratios of terpenes. They are constantly being produced but are vaporized by heat and light of the day.
Terpenes are synthesized in cannabis in secretory cells inside glandular trichomes, and production is increased with light exposure. These terpenes are mostly found in high concentrations in unfertilized female cannabis flowers prior to senescence (the condition or process of deterioration with age). The essential oil is extracted from the plant material by steam distillation or vaporization. Many terpenes vaporize around the same temperature as THC (which boils at about 157°C), but some terpenes are more volatile than others. Terpenes also play an incredibly important role by providing the plant with natural protection from bacteria and fungus, insects and other environmental stresses.
Terpenes are diverse group of organic Hydrocarbons, produced by a wide variety of plants which are important building blocks for certain odors, hormones, vitamins, pigments, steroids, resins, essential oils, and CANNABINOIDS! They are naturally released from plants when temperatures are higher, helping to seed clouds which then cool the plants.
Cannabis is far more complex than only one, or two, cannabinoids delivering the therapeutic effect. Terpenes modify and modulate the effects of THC, CBD, and other cannabinoids and impact the overall medicinal properties of the particular cultivar. Terpenes are also predominant players in the smell and taste of medicinal cannabis. Terpenes interact with neurological receptors; a few bind weakly to Cannabinoid receptors, some seem to alter the permeability of cell membranes and allow in either more or less THC, and others affect serotonin and dopamine chemistry (neurotransmitters).
The Carlini et al study demonstrated that there may be potentiation (a form of synaptic plasticity that is known to be important for learning and memory) of the effects of THC by other substances present in cannabis. The double-blind study found that cannabis with equal or higher levels of CBD and CBN to THC induced effects two to four times greater than expected from THC content alone. The effects of smoking twice as much of a THC-only strain were no different than that of the placebo.
Study done by Wilkinson et al to determine whether there is any advantage in using cannabis extracts compared with using isolated THC. A standardized cannabis extract of THC, CBD and CBN (SCE), another with pure THC, and also one with a THC-free extract (CBD) were tested on a mouse model of multiple sclerosis (MS) and a rat brain slice model of epilepsy Scientists found that SCE inhibited spasticity in the MS model to a comparable level of THC alone, and caused a more rapid onset of muscle relaxation and a reduction in the time to maximum effect than the THC alone.
Most Common Cannabis Terpenes are:
α – Pinene and β - Pinene
Pinenes are a bicyclic monoterpenoid. There are two structural isomers of pinene found in nature: α - pinene and β - pinene. Both forms are important components of pine resin. α - pinene is the most widely encountered terpenoid in nature. Pinenes are used in medicine as an anti-inflammatory, expectorant, bronchodilator and local antiseptic. α - pinene is a natural compound isolated from pine needle oil which has shown anti-cancer activity and has been used as an anti-cancer agent in Traditional Chinese Medicine for many years. It is also believed that the effects of THC may be lessened if mixed with pinene.
Limonene is a monocyclic monoterpenoid and one of two major compounds formed from pinene. As the name suggests, varieties high in limonene have strong citrusy smells like oranges, lemons and limes. Strains high in limonene promote a general uplift in mood and attitude.
Limonene is highly absorbed by inhalation and quickly appears in the bloodstream. It assists in the absorption of other terpenes through the skin and other body tissue. It is well documented that limonene suppresses the growth of many species of fungi and bacteria, making it an ideal antifungal agent for ailments such as toenail fungus. Limonene may be beneficial in protecting against various cancers, and orally administered limonene is currently undergoing clinical trials in the treatment of breast cancer. It is documented that limonene to be; antioxidant, antifungal, anti-bacterial, anti-carcinogenic; good for muscle tension, sleeplessness, pain, inflammation, dissolves gallstones, mood-enhancer, heartburn, and depression.
Plants use limonene as a natural insecticide to ward off predators. It has very low toxicity and adverse effects are rarely associated with it.
β - Caryophyllene
β - Caryophyllene is the only sesquiterpene known to interact with the endocannabinoid system (CB2). Studies show β–caryophyllene holds promise in cancer treatment plans. Research shows shows that β – caryophyllene selectively binds to the CB2 receptor and that it is a functional CB2 agonist. Further, β –caryophyllene was identified as a functional non-psychoactive CB2 receptor ligand in foodstuff and as a macrocyclic anti-inflammatory cannabinoid in cannabis.
Studies show that other phytocannabinoids in combination, especially cannabidiol (CBD) and β - caryophyllene, delivered by the oral route appear to be promising candidates for the treatment of chronic pain due to their high safety and low adverse effects profiles. Other study suggests β - caryophyllene, through a CB2 receptor dependent pathway, may be an excellent therapeutic agent to prevent nephrotoxicity (poisonous effect on the kidneys) caused by anti-cancer chemotherapy drugs such as cisplatin. Another investigative study indicates that the chemical composition of essential oil isolated from black pepper, of which β - caryophyllene is a main constituent, and studied its pharmacological properties. Black pepper oil was found to possess antioxidant, anti-inflammatory and antinociceptive properties. This suggests that high β - caryophyllene strains may be useful in treating a number of medical issues such as arthritis and neuropathy pain.
Linalool is a non-cyclic monoterpenoid and has been described as having floral and lavender undertones. Varieties high in linalool promote calming, relaxing effects.
Linalool has been used for centuries as a sleep aid. Linalool lessens the anxious emotions provoked by pure THC, thus making it helpful in the treatment of both psychosis and anxiety. Studies also suggest that linalool boosts the immune system; can significantly reduce lung inflammation; and can restore cognitive and emotional function (making it useful in the treatment of Alzheimer’s disease).
Studies shown that, linalool may significantly reduce lung inflammation caused by cigarette smoke by blocking the carcinogenesis induced by benz(α)anthracene, a component of the tar generated by the combustion of tobacco. Furthermore, linalool boosts the immune system as it directly activates immune cells through specific receptors and/or pathways. A study suggests linalool may reverse the histopathological (the microscopic examination of biological tissues to observe the appearance of diseased cells and tissues in very fine detail) hallmarks of Alzheimer’s Disease and could restore cognitive and emotional functions via an anti-inflammatory effect. Linalool is a critical precursor in the formation of Vitamin E.
Furthermore, the Environmental Protection Agency has approved its use as a pesticide, flavor agent and scent. Its vapors have been shown to be an effective insecticide against fruit flies, fleas and cockroaches.
Terpinolene is a common component of sage and rosemary and is found in the oil derived from Monterey cypress. It is found most commonly in sativa-dominant strains. Terpinolene’s potential medical benefits include; anticancer, antioxidant, sedative, antibacterial, and antifungal.
Terpinolene has been found to be a central nervous system depressant used to induce drowsiness or sleep or to reduce psychological excitement or anxiety. Further, terpinolene was found to markedly reduce the protein expression of AKT1 in K562 cells and inhibited cell proliferation involved in a variety of human cancers.
Camphene, a plant-derived monoterpene, emits pungent odors of damp woodlands and fir needles. Camphene may play a critical role in cardiovascular disease.
Camphene has been found in various strains of Cannabis sativa. Studies has shown. Camphene reduces plasma cholesterol and triglycerides in hyperlipidemic rats. Given the importance that the control of hyperlipidemia plays in heart disease, the results of this study provide insight into to how camphene might be used as an alternative to pharmaceutical lipid lowering agents which are proven to cause intestinal problems, liver damage and muscle inflammation. This finding alone warrants further investigation.
α - Terpineol, terpinen-4-ol, and 4-terpineol are three closely related monoterpenoids.
The aroma of terpineol has been compared to lilacs and flower blossoms. Terpineol is often found in cannabis varieties that have high pinene levels, which unfortunately mask the fragrant aromas of terpineol.
Terpineol, specifically α - terpineol, is known to have calming, relaxing effects. It also exhibits antibiotic, AChe inhibitor and antioxidant antimalarial properties.
Phellandrene is another terpene that is found in cannabis plants. It is described as pepperminty, with a slight scent of citrus. Phellandrene is believed to have special medicinal values. It has been used in Traditional Chinese Medicine to treat digestive disorders as well as for prevention and treatment of systemic fungal infections.
Δ³ - Carene
Δ³ - Carene is a bicyclic monoterpene with a sweet, pungent odor. It is found naturally in many healthy, beneficial essential oils, including cypress oil, juniper berry oil and fir needle essential oils.
Studies shown, from 162 marihuana plants, which represented over 80 strains. They detected carene in many of the samples.
In higher concentrations, delta-3-carene can be a central nervous system depressant. It is often used to dry out excess body fluids, such as tears, mucus, and sweat.
It is nontoxic, but may cause irritation when inhaled. Perhaps high concentrations of Δ³ - Carene in some strains may be partially responsible for symptoms of coughing, itchy throat and eye afflictions when smoking cannabis.
Humulene is a sesquiterpene also known as α -humulene and α –caryophyllene; an isomer of β–caryophyllene. Humulene is found in hops, cannabis sativa strains, and Vietnamese coriander, among other naturally occurring substances. Humulene is what gives beer its distinct ‘hoppy’ aroma.
Humulene is considered to be anti-tumor, anti-bacterial, anti-inflammatory, and anorectic (suppresses appetite). It has commonly been blended with β–caryophyllene and used as a major remedy for inflammation. Humulene has been used for generations in Chinese medicine. It aids in weight loss by acting as an appetite suppressant.
Pulegone, a monocyclic monoterpenoid, is a minor component of cannabis. Higher concentrations of pulegone are found in rosemary. Rosemary breaks down acetylcholine in the brain, allowing nerve cells to communicate more effectively with one another.
An ethnopharmacology study indicates pulegone may have significant sedative and fever-reducing properties. It may also alleviate the side effects of short-term memory loss sometimes associated with higher levels of THC.
Pulegone has a pleasant peppermint aroma and is considered to be a strong insecticide.
Sabinene is a bicyclic monoterpene found in cannabis. Sabinene aromas are reminiscent of the holidays (pines, oranges, spices). Results of a recent study suggest that sabinene should be explored further as a natural source of new antioxidant and anti-inflammatory drugs for the development of food supplements, nutraceuticals or plant-based medicines.
Sabinene occurs in many plants, including Norway spruce, black pepper, basil and Myristica fragrans (an evergreen indigenous to the Moluccas)—the Spice Islands of Indonesia. The seeds of the Myristica fragrans are the world’s main source of nutmeg. Sabinene exists as (+) and (–) enantiomers.
Geraniol is a monoterpene alcohol that boils at about 447˚F and frequently occurs in cannabis strains that also produce linalool. Geraniol produces a sweet, delightful smell similar to roses. It is known to be an effective mosquito repellant.
Potential medical benefits attributed to geraniol include; antioxidant, anti-tumor, neuroprotectant, anti-bacterial, anti-fungal, anti-viral, and anti-spasmodic.
Here at ARA – Avanti Rx Analytics, we use validated methods using a Gas Chromatography Headspace Analyzer with a Flame Ionization Detector (GC/FID) for the separation and quantification of the following terpenes and terpenoids:
Please note, we only accept samples from authorized; Licensed Producers (LPs), healthcare organizations, and individuals and practitioners who are approved by Health Canada under Access to Cannabis for Medical Purposes Regulations (ACMPR) (i.e., repealed MMPR).
Furthermore we accept cannabis samples for testing and R&D activities from USA who are qualified and approved by DEA (Drug Enforcement Agency) and Health Canada OCS once all the regulatory requirements are approved by both US DEA and Health Canada OCS.
The above process is a Must requirement to ensure compliance with the controlled substance/s chain of custody as per Health Canada OCS regulation.
Please ensure your cannabis Flower/bud sample amount is sufficient for the requested analysis. The amount required for full analysis including pesticides and terpene analysis may require up to about 200g of cannabis. Please ensure your sample is representative of the batch produced.
To avoid cross contamination it will be a good practice that to split your sample into two containers; one for micro analysis (approximately 120 g) and the other for general laboratory testing.
Please ensure your sample/s for testing are: