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CHROMATOGRAPHY


compounds that will be eluted according to their partition behaviour (Fig. 3). Another big advantage of the CPC is the huge range of solvents that can be used: water, alkanes, alcohol, or other commonly used liquids. T e ratio of each solvent depends on the partition coeffi cient of the target compound into the biphasic solvent system. For CPC, the ideal partition coeffi cient is between 0.5 to 3. Within this range, the target compound will be effi ciently elute into the CPC system. T e choice to have an appropriate solvent system assures the separation of the target compounds, contamination-free from other compounds. CPC has advantages over HPLC due to the elimination of silica. Having a liquid stationary phase can have a number of benefi ts. T ere is no non-specifi c adsorption to a solid support, and there is a much higher sample loading capacity as the volume normally taken up by the solid support is occupied by the liquid stationary phase. All of this increases the capacity for higher throughputs, less solvent usage and huge tolerance of extract compounds that can be achieved. Because of this, researchers can use three to fi ve times less solvent with CPC than with traditional methods.[3] Additionally, once fi ltered, the solvent can be recycled for further lowering cost. Preparative isolation of up to four


diff erent major cannabinoids could be achieved by using CPC as the single technique. T e quality of the isolated cannabinoids (>95% pure by HPLC) is suffi cient for many purposes, such as biological testing on a large scale. Additional GC-FID and TLC data supports the purity of the isolated compounds. Gilson scientists injected 5g of crude cannabis oil and manage to extract 205mg with 99% (HPLC) of CBD.[4]


A typical


Fig. 3. How CPC works. The mobile phase (yellow) migrates though a series of cells that contain the stationary phase (blue). The solutes (A, B, and C) in the mobile phase diff use into the cells according to their respective affi nities to the stationary phase


separation using classical LC would have required at least a two-step process over multiple days.[5,6]


T e huge advantage of


using only liquids without any silica support leads to CPC technique recovery of more than 90% of the total weight of these compounds in the original sample, which is greater than the average recovery rate for HPLC. Finally, CPC is not limited to CBD extraction. T e technique is also used in the purifi cation of many others compounds from natural sources, such as [6]-gingerol from ginger (Zingiber offi cinale Roscoe[5]


or


liquiritin from Chinese licorice (Glycyrrhiza uralensis Fisch.).[7]


CPC is not only an eff ective method for extracting CBD from cannabis, but it can also be a great solution to purify other phytocannabinoid components. As the CBD market grows, and demand for CBD-based products increases,


Fig. 1. The chemical structure of cannabidiol


researchers will need a more effi cient way to purify CBD from cannabis for research. CPC uses less expensive reagents and produces a highly pure product more effi ciently than HPLC and fl ash chromatography, which makes it ideal for the extraction step in the production of CBD or other cannabinoids.


REFERENCES


[1] Cannabidiol (CBD) and its analogs: a review of their eff ects on infl ammation, S. Burstein, Bioorg. Med. Chem; Vol.23, 2015 [2] Multiple mechanisms involved in the large-spectrum therapeutic potential of cannabidiol in psychiatric disorders, Campos et al.; Philos. Trans. R. Soc. B Biol. Sci., Vol. 367, 2012 [3] Evolution of the Cannabinoid and Terpene Content during the Growth of Cannabis sativa Plants from Diff erent Chemotypes; Oier Aizpuruia-Olaizola et al.; J Nat Prod. Vol. 79, No. 2, 2016 [4] CPC 250 PRO Purifi cation of Cannabidiol from Cannabis sativa; Gilson Applications Laboratory. [5] Caulerpenyne from Caulerpa taxifolia: A comparative study between CPC and classical chromatographic techniques; Estell Sfecci et al.; Phytochem Lett. Vol. 20, 2017 [6] Eff ective Cannabinoid Purifi cation by Flash Chromatography; John R. Bickler and Elizabeth Denton; Biotage. Presented at ACS 2016, Philadelphia, PA, 2016. [ 7] Orthogonal Analysis Underscores the Relevance of Primary and Secondary Metabolites in Licorice; Simmler et al. J Nat Prod. Vol.77, 2014


Gregoire Audo works in the application laboratory department at Gilson Purification. www.gilson.com


www.scientistlive.com 31


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