Liquid chromatography is considered an important method for determining concentrations of cannabinoids within the cannabis plant. High performance liquid chromatography (HPLC) is probably the most commonly used method for the separation (or remediation) of THC from CBD. It offers the ability to test for THCA, a compound normally untraceable in other methods. It is also a reliable choice for potency determination, allowing you to identify cannabinoids with similar structures in just one accurate analysis.
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As we have seen, the pre-treatment of CBD before its purification is extremely important in order to provide cannabidiol-based treatments. Our range of Chromalite chromatographic resins offer an affordable and effective method to separate THC from CBD. The chemical and physical strength of these compounds means that traditional silica media can be damaged during chromatography, resulting in the need for expensive replacements. After vigorous testing of their mechanical stability, Chromalite resins show a much lower breakdown over time than equivalent competitor resins. As a result, they are stable for a long lifetime and will withstand tough conditions of use.
Reverse Phase Chromatography
Reverse phase chromatography is a widely utilized analytical method for purification because of its ability to separate compounds of varying hydrophobicities. This is effective in the separation of tetrahydrocannabinol from CBD, as THC has a greater hydrophobicity, allowing efficient removal of small amounts of THC from a CBD-dominant product. The larger the separation peaks identified in this method, the better the purification will be from a higher number of sample loads.
CBD, the colloquial term for Cannabidiol, is one of at least 100 active compounds (cannabinoids) that can be found in hemp or marijuana cannabis plants. These chemical compounds join to the cannabinoid receptors of the body and brain, also known as the endocannabinoid system, to produce different effects.
Chromatography is proving the most effective method in the purification of cannabidiol as it acts as the cleaning stage once the CBD has been extracted. But achieving highly pure CBD for medicinal applications requires numerous pre-treatment stages to ensure the removal of impurities that could potentially foul the resins used in chromatography. This can include shredding the plant, drying, extraction, decarboxylation and distillation before the chromatography step can be performed. The removal of contaminants from cannabinoids is a prominent manufacturing challenge, but chromatography is becoming the leading purification method because of its quick and evident results.
The legalization of cannabis for both medicinal and recreational purposes is a hot industry topic in 2020, as pros and cons of its de-criminalization continue to be debated across the globe. The discovery of specific cannabinoids, such as CBD, that demonstrate promising characteristics for medical treatments, has resulted in an increasing demand for highly purified, safe cannabis-derivatives. This has prompted a need for robust analytical methods of purification that can provide reliable results for both manufacturers and consumers alike.
HPLC Method Parameter:
Ultraviolet Detector: Gelai UV-3292 (wavelength 190-400nm, with preparative flow cell)
Column: Gelai 25.4mm x 450mm SAC Column
a leader in preparative HPLCs
The most prevalent cannabinoids in cannabis are THC, CBD, and THCA. The variety of uses for cannabis also lead to a variety of reasons that the separation of cannabinoids is an important process. One example is the United States classification of industrial hemp as containing no more than 0.3% THC by dry weight, resulting in it not being federally legal to ship or trade product with THC across state lines within the United States. Another is the medicinal use of cannabis, where isolation of CBD and THC are important for the pharmaceutical sector to evaluate the effect of cannabinoids on the human body. The recreational drug use of cannabis has also led to increasing desires to provide a purified product.
95% to 78% B from 70 to 71 minutes
78% to 95% from 66 to 67 minutes
60% B from 0 to 5 minutes
The speed at which regulations have changed in the cannabis marketplace has increased the pressure for accurate, easy, and efficient analytical methodology. Purified isolates are used to formulate a variety of consumer products such as topicals, edibles, and beverages. Accuracy of the quantitative cannabinoid content of the isolates used in preparation of these products is directly dependent upon the reliability of the analytical methodology used for characterization. An inaccurate quantitative assay value does not only pose health, safety, and ethical concerns for consumers, it can generate a potential financial loss for manufacturers marketing isolates based upon purity. The aim of this work is to describe the validation of a rapid and reliable reversed-phase liquid chromatographic method coupled with UV detection for quantitative determination of Cannabidiol (CBD) purified isolate. Accuracy, precision, specificity, linearity, robustness and range of the analytical method were determined according to assay method requirements specified in the International Conference for Harmonization (ICH) Q2(R1), “Validation of Analytical Procedures: Test and Methodology” for quantitative evaluation of the active moiety in samples of drug substance by liquid chromatography. Additionally, the feasibility of manufacturer prepared DEA-exempt CBD reference standard solutions formulated at 1.0 mg/mL was explored for the quantification of CBD isolate.