Separation of cannabinoids for clinical use

PrepChrom C-700: Isolation of cannabidiol (CBD) and tetrahydrocannabinol (THC) from other cannabinoids extracted from cannabis

PrepChrom WordPressThe separation of cannabinoids is an important process which is necessary for clinical studies. The aim of these studies is to investigate the effect of the isolated cannabinoids on patients suffering from chronic illnesses and side effects caused by chemotherapy. The relaxation of laws concerning medical use of cannabis all around the world, especially the United States, has increased the interest in determining the effect of cannabinoids on the human body. Clinical studies require large amount of highly pure separated cannabnnoids. This can easily be achieved with the PrepChrom C-700.

Introduction

Cannabis is an extremely versatile plant and has been used for many purposes throughout history. Some specific sorts are used for fibers in the textile and paper industry because of their long stems. It is a useful source of foodstuffs, such as hemp oil and seed. Due to the cannabinoids, cannabis is also a popular recreational drug. These substances can cause mental and physical effects when consumed. The impact of the cannabinoids on the human body is of great interest for the pharmaceutical sector. The research emphasis lies in treating the side effects of chemotherapy, inflammatory diseases like multiple sclerosis or degenerative illnesses such as Parkinson’s disease.[3] AI FAME GmbH, a Swiss company, is a pioneering company to extract the plant-based active substances and make them water-soluble for improved, further processing.

Here, we aim to separate three cannabinoids, i.e. cannabidiol, tetrahydrocannabinol and tetrahydrocannabinolic acid in high purity from the extract.

Experimental

Equipment: PrepChrom C-700

Sample: Water-soluble cannabis extract. Cultivated, extracted and provided by AI FAME GmbH

Preparation: The chromatographic separation is performed with 1 mL of the water-soluble cannabis extract diluted in 1 mL methanol : water (1:1)

Separation: Method parameters
Column:  Sepacore® C-18 80 g
Particle size:  40-63 μm
Flow rate:  40 mL/min
Sample loop:  2 mL
Detection:  253, 270 nm and SCAN 200-600 nm

Gradient: methanol (A) : water(B)
50 % – 90 % methanol (A) in 20 min
90 % – 90 % methanol (A) in 10 min
90 % – 95 % methanol (A) in 5 min
95 % – 95 % methanol (A) in 10 min

Results

Cannabidiol (CBD) bearing a 1,3-diol functional group on its benzene ring, is the first substance to be eluted when applying reversed phase conditions (Figure 1), hence, is the most polar cannabinoid in the sample. The second compound, eluting after 27 minutes, is tetrahydrocannabinol (THC). The last substance to be eluted is tetrahydrocannabinolic acid (THCA), a form of cannabinoid which easily decarboxilates in to THC when exposed to light or heat. Assignment of the signals and identification was done by comparison with reference HPLC data provided by AI FAME GmbH.

Cannabinoide SN finalFigure 1: Methanol : water separation of the extracted cannabinoids using the PrepChrom C-700. Detection at 270 nm, 254 nm, and 200-600 nm SCAN.

 

 

Conclusion

The three cannabinoids extracted by AI FAME GmbH could easily be separated and separately collected with the PrepChrom C-700.

The water-solubility of the extract enables the use of aqueous solvents that have inherent economic and ecologic advantages over common organic solvents.

The growing interest in the clinical use of cannabinoids is unbreakable. Furthermore, cannabis is decriminalized and it is allowed to exploit the potential of its valuable substances. BUCHI offers straight forward solutions to separate and purify small to large size batches of the precious extracts.

Acknowledgment

AI FAME GmbH, Switzerland, is thanked for providing the extract and for the fruitful analytical discussion.

References

[1] Sutton, IR. Daeniken, P. (2006). Cannabinoids in the management of intractable chemotherapy-induced nausea and vomiting and cancer-related pain. The Journal of Supportive Oncology.

[2] State Medical Marijuana Laws. National Conference of State Legislatures. (16.3.2015). http://www.ncsl.org/research/health/state-medical-marijuana-laws.aspx

[3] Kogan, NM. Mechoulam, R. (2007) Cannabinoids in health and disease. Dialogues Clin Neurosci. 2007 Dec; 9(4): 413–430.

>> See additional Application Notes

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NEW Product Announcement

BUCHI PrepChrom C-700: bridging the gap between flash chromatography and preparative HPLC

BUCHI introduces the innovative PrepChrom C-700 chromatography system, combining flash chromatography with preparative HPLC. Bridging the gap between them, the C-700 sets a new standard for purification of synthesis mixtures or complex natural extracts.

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The space saving and easy to use system allows organic chemists to perform the complete purification process, using the interactive user interface specially designed for preparative chromatography applications.

The high quality pumping system features a quaternary gradient module and delivers accurate flow rates up to 250 mL/min at a pressure up to 100 bar (1450 psi). Any type of flash column as well as preparative HPLC columns with a diameter up to 50 mm filled with silica having a particle size down to 10 µm can be used.

Combining high-end hardware technology together with the software designed for easy purification, the PrepChrom C-700 is the ideal solution to solve any purification bottleneck in the research of new active compounds.

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Live Webinar: From TLC to Flash to Prep-HPLC on BUCHI’s new all-in-one solution – PrepChrom C-700

07_313345-1038

Wednesday, June 18, 2014
11:00am Eastern Time


What if there were a way to combine flash chromatography and preparative HPLC into one solution that took up a quarter of the space? Learn about how you can go from TLC to purified product without multiple instruments and without being an expert in chromatography. A test application will show how one product can give you the results you need quickly and easily.

Key Learning Objectives:
• Learn about your options to purify compounds and reaction mixtures.
• Learn how several techniques involved in preparative chromatography can be combined into one solution.
• See interesting applications optimized from TLC to flash and from flash to HPLC fractions.

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Sepacore Easy Purification Systems: better and faster separation

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By: Brigitte Pichon

BUCHI introduces the new Sepacore Easy Purification Systems. These preparative chromatography start-up packages enable the purification required after a chemical synthesis or the extraction of a natural product. As a first step toward automation, they allow to precisely control the flow rate and the eluent composition of the mobile phase. Compared with the use of an open glass column, the Easy Purification Systems enable an improved separation with a decreased purification cost.

The “Easy Synthesis” system featuring a 250 mL/min, 10 bar (145 psi) pumping system offers an ideal configuration for the purification of synthesis mixtures on flash cartridges. Compared with the manual purification, the accurate control of the gradient elution provides an increased resolution and therefore a higher purity of the collected fractions.

The “Easy Extract” system is designed for large sample loading such as natural extracts. For such applications, the 50 bar (725 psi) gradient system is able to feed large glass columns for the purification of several hundred grams of natural extract. Together with a controlled separation process, such a system also allows to significantly reduce the solvent consumption when compared with the manual feeding of the solvents on an open glass column.

Both systems can later on easily be upgraded to a completely automated system including UV and/or ELS detection, fraction collection and flexible computer control.

For more information visit: http://purification-systems.buchi.com

Universal detection with the new ELS Detector C-650

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The Evaporative Light Scattering Detector (ELSD) C-650 detects all non-volatile compounds, including those which cannot be detected using UV absorption. It can handle any combination of mobile phases and gradients required for efficient and fast separations. A proprietary flow splitter provides a controlled flow rate to feed the ELSD for any application requiring a flow rate up to 250 mL/min.
When used in parallel with the C-640 UV-Vis Detector, it allows monitoring all the compounds eluting from the column and collecting them accordingly.
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Flash Chromatography Note #61: Isolation of 3-Nitro-4-ethoxybenzaldehyde

Buchi Flash Chromatography Sepacore System

Buchi Flash Chromatography Sepacore System

This flash chromatography short note describes a simple and fast procedue for the isolation of 3-Nitro-4-ethoxybenzaldehyde from a synthesis mixture. Visit our Sepacore Flash Chromatography Applicationssection to download this short note and review all our other available application studies.

Sepacore Configuration:
Cartridge 12 x 150mm, prepacked with silica gel 60, 40 – 63 μm
2 Pump Modules C-605
Fraction Collector C-660
Control-Unit C-620 with SepacoreControl software
UV Photometer C-635

First Separation Conditions:
Eluent: n-hexane with 25 / 50 / 75% ethyl acetate (%B), step gradient
Flow rate: 10ml/min
Sample: 0.5 g crude mixture, dissolved in n-hexane/ethyl acetate 75:25
Injection volume: 1ml

Under these conditions the impurity with the shortest elution time cannot be separated from the main component. Therefore the solvent gradient was modified by decreasing the content of the more polar solvent (ethyl acetate) in the first step.

Second Separation Conditions:
Eluent: n-hexane with 15 / 50 / 75% ethyl acetate (%B), step gradient