GB2567235A

GB2567235A - Ozonation of cannabinoids

Info

Publication number: GB2567235A
Application number: GB1716491.4A
Authority: GB
Inventors: Daniel Mayers Michael
Original assignee: Individual
Current assignee: Individual
Priority date: 2017-10-09
Filing date: 2017-10-09
Publication date: 2019-04-10
Anticipated expiration: 2037-10-09
Also published as: GB201716491D0; GB2567235B

Abstract

A method of ozonating a non-psychoactive cannabinoid by providing a source of non-psychoactive cannabinoid and contacting it with ozone. By contacting the non-psychoactive cannabinoid with ozone, it is converted to a psychoactive cannabinoid. In one aspect of the invention, the psychoactive cannabinoid is then isolated. The non-psychoactive cannabinoid is preferably cannabidiol (CBD), cannabinol (CBN), or cannabichromene (CBC), and the source is preferably hemp, hemp chaff, or hempseed oil. The psychoactive cannabinoid produced is tetrahydrocannabinol (THC), either Δ9-tetrahydrocannabinol or Δ8-tetrahydrocannabinol. The method of ozonating the cannabinoid is preferably in an enclosed space such as a sealed compartment. The ozone is preferably provided by an ozone generator. A composition containing the psychoactive cannabinoid produced may be used for treatment of pain, or some ailments and diseases.

Description

(57) A method of ozonating a non-psychoactive cannabinoid by providing a source of non-psychoactive cannabinoid and contacting it with ozone. By contacting the non-psychoactive cannabinoid with ozone, it is converted to a psychoactive cannabinoid. In one aspect of the invention, the psychoactive cannabinoid is then isolated. The nonpsychoactive cannabinoid is preferably cannabidiol (CBD), cannabinol (CBN), or cannabichromene (CBC), and the source is preferably hemp, hemp chaff, or hempseed oil. The psychoactive cannabinoid produced is tetrahydrocannabinol (THC), either A⁹-tetrahydrocannabinol or A⁸-tetrahydrocannabinol. The method of ozonating the cannabinoid is preferably in an enclosed space such as a sealed compartment. The ozone is preferably provided by an ozone generator. A composition containing the psychoactive cannabinoid produced may be used for treatment of pain, or some ailments and diseases.

OZONATION OF CANNABINOIDS

FIELD OF THE INVENTION

The present invention relates generally to the field of chemical synthesis, in particular ozonation reactions of cannabinoids. More specifically, the present invention relates methods of converting non-psychoactive cannabinoids to psychoactive cannabinoids.

BACKGROUND OF THE INVENTION

Recently, public interest in Cannabis as medicine has been growing, based in no small part on the fact that Cannabis has long been considered to have medicinal properties, ranging from treatment of cramps, migraines, convulsions, appetite stimulation and attenuation of nausea and vomiting. In fact, a report issued by the National Academy of Sciences’ Institute of Medicine indicated that the active components of Cannabis appear to be useful in treating pain, nausea, AIDS related weight loss or “wasting”, muscle spasms in multiple sclerosis as well as other problems. Advocates of medical marijuana argue that it is also useful for glaucoma, Parkinson’s disease, Huntington’s disease, migraines, epilepsy and Alzheimer’s disease.

Marijuana refers to varieties of Cannabis having a high content of A⁹-tetrahydrocannabinol (A⁹-THC), which is the psychoactive ingredient of marijuana, whereas industrial hemp refers to varieties of the Cannabis plant that have a low content of A⁹-THC. Furthermore, A⁹-THC is only one of a family of over a hundred bi- and tri-cyclic compounds named cannabinoids. For example, A⁸-THC is a double bond isomer of A⁹-THC and is a minor constituent of most varieties of Cannabis (Hollister and Gillespie, 1972, Clin Pharmacol Ther 14: 353). The major chemical difference between the two compounds is that A⁹-THC is easily oxidized to cannabinol (CBN), whereas A⁸-THC does not and is in fact very stable. A⁸-THC, for the most part, produces similar psychoactive effects as does A⁹-THC, but is generally considered to be 50% less potent than A⁹-THC and has been shown in some cases to be 3-10 times less potent. A⁸-THC has also been shown to be more (200%) effective an anti-emetic than Δ⁹THC and has been used as an anti-emetic in children, based on the belief that the side effects of A⁹-THC and A⁸-THC, such as anxiety and dysphoria, are more prevalent in adults than children (Abrahamov et al, 1995, Life Sciences 56: 2097-2102).

On the other hand, cannabidiol (CBD) is non-psychoactive and has no activity on its own when administered to humans. It is of note that CBD is typically about 2% (0.5-4%) dry weight of hemp chaff, Δ⁸-ΤΗ0 is approximately 0.2% (0.05-0.5%) dry weight and Δ⁹-ΤΗ0 is approximately 0.1% (0.05-0.3%). Gaoni and Mechoulam (1966, Tetrahedron 22: 1481-1488) teach methods of converting CBD to, among other compounds, Δ⁸-ΤΗΟ and Δθ-THC comprising boiling a solution of CBD (3.0 g) in absolute ethanol (100 ml) containing 0.05% HCI for 18 hours. The solution was then poured into water and extracted with ether. The ether solution was washed with water, dried (Na2SO4) and evaporated. Δ⁸-ΤΗΟ and Δθ-THC were eluted from the resulting oil and separated by chromatography. In another experiment, CBD (3.14 g) was dissolved in benzene (100 ml) containing 2 mg/ml p-toluenesulphonic acid and boiled for two hours. The reaction mixture was poured into Water and the upper layer was separated, washed with 5% NaHCO3, then with water, dried and evaporated. Elution With pentane-ether (95:5) gave an oily material which was subsequently distilled.

Percentage yield of Δ⁸-ΤΗΟ was given as 64% of the crude material in this paper. The crude oil product, which showed only one spot by thin layer chromatography, was purified by vacuum distillation. Gaoni and Mechoulam (1964, J Amer Chem Soc 86: 1646) also described a method for converting CBD to Δθ-THC comprising boiling a mixture of CBD in ethanol containing 0.05% hydrogen chloride for 2 hours. Percentage yield of Δθ-THC was 2% (Mechoulam et al, 1972, J Amer Chem Soc 94: 6159-6165; Mechoulam and Gaoni, 1965, J Amer Chem Soc 87: 3273). Using boron trifluoride, the yield was 70% (Gaoni and Mechoulam, 1971, J Amer Chem Soc 93: 217-224) although purity was not given.

US 7,399,872 describes methods for producing high yield and high purity Δθ-THC or Δ⁸THC, in particular a method of converting CBD to Δθ-THC comprising: providing a reaction mixture comprising CBD in dry methylene chloride; adding BF₃Et₂O to the reaction mixture; stirring the ice cold reaction mixture under a nitrogen atmosphere; adding NaHCO₃ to the reaction mixture; allowing the mixture to separate into an aqueous phase and an organic phase; removing the organic phase; washing the organic layer with water; and eluting Δ⁹THC from the organic phase, the purity of the eluted Δ⁹-ΤΗΟ being greater than 97%.

Clearly, as cannabinoids are of potential medicinal value, improved methods of converting non-psychoactive cannabinoids such as CBD to psychoactive cannabinoids such as Δ⁹-ΤΗΟ or Δ⁸-ΤΗΟ are needed.

SUMMARY OF THE INVENTION

According to a first aspect of the invention, there is provided a method of ozonating a non psychoactive cannabinoid, said method comprising the steps of: a) providing a source of non-psychoactive cannabinoid; and b) contacting the source of non-psychoactive cannabinoid with ozone or a reactive equivalent thereof.

According to a second aspect of the invention, there is provided a method of converting a non-psychoactive cannabinoid to a psychoactive cannabinoid, said method comprising the steps of: a) providing a source of non-psychoactive cannabinoid; b) contacting the source of non-psychoactive cannabinoid with ozone; and c) optionally isolating the psychoactive cannabinoid.

According to a third aspect of the invention, there is provided a psychoactive cannabinoid or mixture of psychoactive cannabinoids obtainable by a method described herein, compositions comprising same, and their use in medicine.

According to a fourth aspect of the invention, there is provided a method of treating one or more of pain, chronic pain, acute pain, cramps, migraine, convulsions, stress, distress, anxiety, nausea, chemotherapy-induced nausea, vomiting, AIDS-related weight loss, wasting, spasm, multiple sclerosis, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, glaucoma, epilepsy and cancer, said method comprising administering the composition described herein to a subject in need thereof.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned hereunder are incorporated herein by reference.

One hundred and thirteen different cannabinoids are found in the Cannabis plant. These can be broadly divided by the person skilled in the art into those which are “psychoactive” and those which are “non-psychoactive”. The major non-psychoactive cannabinoid is cannabidiol (CBD). Cannabinol (CBN) and cannabichromene (CBC) are other less abundant examples of non-psychoactive cannabinoids. Hemp (e.g. industrial hemp) is a variety of the Cannabis sativa plant and is one example of a source of non-psychoactive cannabinoids. In one embodiment of the invention, the ratio of CBD: CBN: CBC is substantially the same as that in natural or industrial hemp.

The most well-characterized psychoactive cannabinoids are the tetrahydrocannabinols, such as A⁸-tetrahydrocannabinol (A⁸-THC) and A⁹-tetrahydrocannabinol (A⁹-THC). Isoforms of Δ⁸' THC according to the invention include Δ⁸ isoTHC and trans- Δ⁸ isoTHC. Δ⁹- and A⁸-and tetrahydrocannabinol (A⁹-THC) are preferred embodiments of the present invention.

Ozone (or trioxygen) is an inorganic molecule with the chemical formula O₃. As used herein, “ozonation” refers to the contacting, treatment or chemical reaction of an agent or substance with ozone. For example, the ozone gas may be bubbled or infused into a liquid. Alternatively, the ozone gas may be allowed to diffuse and contact a solid. The ozone may contact other gases. In a preferred embodiment of the present invention, the ozone is provided by an ozone-generating device. Suitable devices are known to the person skilled in the art and include the BESTEK 03 ozone generator/air sterilizer model: BTAS807.

In one embodiment of the invention, the step of contacting the non-psychoactive cannabinoid with ozone is carried out in an enclosed space, preferably in a substantially or hermetically sealed compartment. The compartment may be a bag or box. The sealing or compartmentalisation preferably allows sufficient concentration of ozone to build up in the reaction space, and prevents diffusion of the ozone away from the general vicinity of the reaction. In a related embodiment, the non-psychoactive cannabinoid is immobilised on a support.

In one embodiment, the non-psychoactive cannabinoid is converted to at least one psychoactive cannabinoid. The psychoactive cannabinoid may be isolated or purified from the reaction mixture e.g. by means known in the art and as described herein.

Also provided herein are methods and protocols for converting non-psychoactive cannabinoids such as cannabidiol (CBD) to psychoactive cannabinoids such as Δ⁸tetrahydrocannabinol and/or A⁹-tetrahydrocannabinol. In one embodiment, CBD is converted to an ozonated psychoactive cannabinoid. In one embodiment, CBD is converted to a psychoactive ozonated derivative of CBD. As will be appreciated by one knowledgeable in the art and as discussed below, the reaction times may be varied somewhat, producing product at different yields and purities. Furthermore, functional equivalents may be substituted where appropriate. For example, the conversion reaction may be carried out from anywhere between 30 minutes to 24 hours, such as 1 hour to 24 hours, such as about 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours. In a preferred embodiment, the conversion is carried out for about 1.5 hours.

The psychoactive cannabinoid may be processed into a composition. For example, the psychoactive cannabinoid may be combined with suitable excipients known in the art, thereby forming a pharmaceutical composition. In some embodiments, the psychoactive cannabinoid at therapeutically effective concentrations or dosages may be combined with a pharmaceutically or pharmacologically acceptable carrier, excipient or diluent, either biodegradable or non biodegradable. Exemplary examples of carriers include, but are by no means limited to, for example, poly(ethylene-vinyl acetate), copolymers of lactic acid and glycolic acid, poly(lactic acid), gelatin, collagen matrices, polysaccharides, poly(D,L lactide), poly(malic acid), poly(caprolactone), celluloses, albumin, starch, casein, dextran, polyesters, ethanol, methacrylate, polyurethane, polyethylene, vinyl polymers, glycols, mixtures thereof and the like. Standard excipients include gelatin, casein, lecithin, gum acacia, cholesterol, tragacanth, stearic acid, benzalkonium chloride, calcium stearate, glyceryl monostearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyethylene glycols, polyoxyethylene stearates, colloidol silicon dioxide, phosphates, sodium dodecylsulfate, carboxymethylcellulose calcium, carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethycellulose phthalate, noncrystalline cellulose, magnesium aluminum silicate, triethanolamine, polyvinyl alcohol, polyvinylpyrrolidone, sugars and starches. See, for example, Remington: The Science and Practice of Pharmacy, 1995, Gennaro ed.

The route of administration of the composition of the invention may be by any known in the art. Particularly preferred are the oral and intrapulmonary route, e.g. by inhalation. Via the oral route, the product may be first mixed with butter or an edible oil.

In some embodiments of the process of the invention, yield of product is at least 50%. In other embodiments, the yield is at least 60%. In other embodiments, yield is at least 70%. In yet other embodiments, yield is 70-85%. Yield is determined by looking at the peak area for the isolated compound in the gas chromatography-mass spectra analysis of the crude reaction product mixture. Purity is also determined by GC-MS and also by analytical HPLC. The total ion chromatogram from the GC-MS gives information similar to that provided by an FID-GC in that the peak area is proportional to the mass of the analytes detected. Total peak area and the peak areas of the individual analytes can be compared in the GC-MS case as long as the masses are in generally the same range. As discussed below, in some embodiments, purity of the tetrahydrocannabinols isolated by the process is greater than 90%. In yet other embodiments, purity is greater than 95%. In yet other embodiments, purity is greater than 97% or greater than 98%. In yet other embodiments, purity is 98-99%.

In an additional aspect, the present invention provides a method of converting cannabidiol (CBD) to A⁹-tetrahydrocannabinol (Δθ-THC) comprising providing a reaction mixture comprising CBD in dry methylene chloride; adding BF3Et2O to the reaction mixture; stirring the ice cold reaction mixture under an ozone atmosphere; adding NaHCO³ to the reaction mixture; allowing the mixture to separate into an aqueous phase and an organic phase; removing the organic phase; washing the organic layer with water; and eluting Δθ-THC from the organic phase, the purity of the eluted ΔΘ-THC being greater than 97%, such as greater than 98%.

In this aspect, the reaction mixture may be stirred for approximately 1 hour. The method may include drying the organic phase over MgSO₄ and evaporating the organic phase following washing. The method may comprise eluting the organic phase on an HPLC column. The Δθ-THC may be eluted with ether in petroleum ether following washing the column with petroleum ether. The ether in petroleum ether may be 2 parts ether in 98 parts petroleum ether. The method may comprise eluting the organic phase on an RP-HPLC column.

Specifically, described herein is a method of converting CBD to a tetrahydrocannabinol comprising: providing a reaction mixture comprising a catalyst in an organic solvent, adding CBD to the reaction mixture, mixing said reaction mixture under ozone, allowing the mixture to separate into an aqueous phase and an organic phase; removing the organic phase, and eluting the tetrahydrocannabinol from the organic phase.

As used herein, “Lewis acid” refers to a powerful electron pair acceptor. In some embodiments, the catalyst is a Lewis acid, for example, p-toluenesulfonic acid, boron trifluoride or BF₃Et₂O. In some embodiments, the BF₃Et₂O is in dry methylene chloride, ethyl acetate, ethanol, hexane or other organic solvent. In yet other examples, the catalyst may be hydrochloric acid in ethanol or sulphuric acid in cyclohexane. In some embodiments, a weak base is added to the reaction mixture prior to allowing the reaction mixture to separate into organic and aqueous phases. The base may be an alkali metal hydrogen carbonate or a carbonate of an alkali metal. In some embodiments, the organic layer is dried prior to eluting. In these embodiments, a suitable drying or dehydration compound, for example, MgSO₄ or Na₂SO₄ is used. In yet other embodiments, the process may be carried out under a nitrogen atmosphere.

The invention will now be described by means of examples, although the invention is not limited to these examples.

EXAMPLE I

A quantity of dry hemp chaff was placed in a bag together with an operative ozonegenerating machine (BESTEK 03 ozone generator/air sterilizer model: BTAS807) for approximately 1.5 hours. A sample of each of the treated and the untreated hemp was pulverized and prepared for smoking using standard methods known in the art. The sample was smoked by 15 professional smokers trained to recognise a psychoactive effect prior to the experiment. 14 out of 15 panellists confirmed a psychoactive effect of the treated sample but not the untreated sample. The sample is further subjected to NMR/IR spectroscopy and presence of Δθ-THC and A⁸-THC, as well as other ozonated cannabinoid products confirmed.

EXAMPLE II

Ozonation of hemp was carried out as per Example I above. The ozonated product was sampled by a subject suffering from chronic pain and anxiety. The product was firstly smoked by the subject. The subject confirmed amelioration of symptoms, in particular lessening of pain. On a separate occasion, the subject mixed the product with edible oil and ingested the product. The subject again confirmed amelioration of symptoms, but more long-lasting than when smoked.

EXAMPLE III

Ozone was produced from oxygen under electric discharge conditions. Before being fed into the ozone generator, oxygen was dried with Mg(CIO₄)₂. The solvent was CCI₄ (reagent grade). Prior to performing the reaction, substrate (hempseed oil) was treated with an ozone-oxygen mixture (~10% O₃) for 30 min for decomposing the possible active impurities, held over calcium chloride, and distilled. Ozonation was carried out in a temperaturecontrolled bubbler under steady-state conditions. The reaction was performed in the temperature-controlled sample cell of a spectrophotometer. An ozone-oxygen mixture was passed through the sample cell with a substrate solution until the necessary O₃concentration in the solution (1.7 χ 10^-5 to 6.8 χ 10^-4 mol/l) was reached. The blank in the reference cell was a solution containing the same substrate concentration. The substrate and ozone concentrations in the sample cell were chosen so that the substrate was at least in 100-fold excess over ozone. After the necessary O₃ concentration was reached, the sample cell was sealed hermetically and the ozone consumption rate in the cell was monitored spectrophotometrically. Using this procedure, we determined the monomolecular ozone disappearance rate constant (k_mono). From the dependence of k_mono on the substrate concentration, it is possible to determine the stoichiometry of the reaction and the ozonation rate constant at the given temperature.

EXAMPLE IV

Conversion of CBD to A⁸-THC. CBD (300 mg) was added to dried p-toluenesulfonic acid (30 mg) in toluene (15 ml), under O₃ atmosphere. In this example, the mixture was refluxed (under O₃) for 1 hour, although other time periods may also be used, as discussed below. It was then diluted with ether (20 ml) and poured into cold water. The upper layer was separated, washed with aqueous 5% NaHCO₃, then with water, dried over MgSO₄ and evaporated. The viscous oil showed mainly one spot on TLC (using 20% ether in petroleum ether as eluent). HPLC, on the crude oil, showed the presence of 87% Δ⁸-ΤΗ0. The oil was chromatographed on a silica gel column (6 g). Elution with 5 to 10% ether in petroleum ether gave a fraction (244 mg, 81%) of A⁸-THC 98.8% pure. When the reaction was carried out using various reflux times showed the presence of 79.37% A⁸-THC (15 minutes), 81.9% Δ⁸THC (30 minutes) and 84.7% A⁸-THC (2 hours).

EXAMPLE V

Conversion of CBD to Δθ-THC. BF₃Et₂O (50 ul) was added, under ozone atmosphere, to ice cold solution of CBD (300 mg) in dry methylene chloride (15 ml). The solution was stirred at 0° C for 1 hour. Saturated aqueous solution of NaHCO₃ (2 ml) was added until the red colour faded. The organic layer was removed, washed with water, dried over MgSO₄ and evaporated. The composition of the oil obtained (determined by HPLC): trans- A⁸-isoTHC 27%, Δθ-THC 69.7%. The oil was chromatographed on silica gel column (20 g) and eluted with petroleum ether followed by graded mixtures, up to 2:98 of ether in petroleum ether. The first fraction eluted was the A⁸-isoTHC (30 mg, 9.5%) followed by a mixture of A⁸-iso THC and Δθ-THC (100 mg). The last compound to be eluted was the Δθ-THC (172 mg, 57%). The purity of Δθ-THC (as determined by HPLC) Was 98.9%. It is of note that when the reaction was carried in the presence of MgSO₄ (120 mg), the composition of the oil obtained (determined by FIPLC) was: trans- A⁸isoTHC 21.15%, Δθ-THC 56.7%.

While the preferred embodiments of the invention have been described above, it will be recognised and understood that various modifications may be made therein, and the appended claims are intended to cover all such modifications which may fall within the spirit and scope of the invention.

Claims

1. A method of ozonating a non-psychoactive cannabinoid, said method comprising the steps of:

a) providing a source of non-psychoactive cannabinoid;

b) contacting the source of non-psychoactive cannabinoid with ozone or a reactive equivalent thereof.

2. The method according to claim 1, wherein the non-psychoactive cannabinoid is selected from the group consisting of: cannabidiol (CBD), cannabinol (CBN) and cannabichromene (CBC).

3. The method according to claim 1 or claim 2, wherein the non-psychoactive cannabinoid is cannabidiol (CBD).

4. The method according to any one of claims 1 to 3, wherein the source of nonpsychoactive cannabinoid is hemp, hemp chaff or hempseed oil.

5. The method according to any one of claims 1 to 4, wherein the contacting step is carried out in an enclosed space, preferably in a substantially or hermetically sealed compartment.

6. The method according to any one of claims 1 to 5, wherein the ozone is provided by an ozone-generating device.

7. The method according to any one of claims 1 to 6, wherein the contacting step is carried out for between about 1 to about 24 hours.

8. The method according to claim 7, wherein the contacting step is carried out for about 1.5 hours.

9. The method according to any one of claims 1 to 8, wherein the non-psychoactive cannabinoid is converted to at least one psychoactive cannabinoid.

10. The method according to claim 9, wherein the psychoactive cannabinoid is a tetrahydrocannabinol.

11. The method according to claim 9 or claim 10, wherein the psychoactive cannabinoid is A⁸-tetrahydrocannabinol and/or A⁹-tetrahydrocannabinol.

12. The method according to any one of claims 9 to 11, further comprising the step of isolating the psychoactive cannabinoid.

13. A method of converting a non-psychoactive cannabinoid to a psychoactive cannabinoid, said method comprising the steps of:

a) providing a source of non-psychoactive cannabinoid;

b) contacting the source of non-psychoactive cannabinoid with ozone;

c) optionally isolating the psychoactive cannabinoid.

14. The method according to claim 13, wherein the non-psychoactive cannabinoid is cannabidiol and/or the psychoactive cannabinoid is a tetrahydrocannabinol or a mixture of tetrahydrocannabinols.

15. A psychoactive cannabinoid or mixture of psychoactive cannabinoids obtainable by the method of any one of claims 1 to 14.

16. A composition comprising the psychoactive cannabinoid or mixture of psychoactive cannabinoids of claim 15.

17. The composition of claim 16 for use in medicine.

18. The composition of claim 16 for use in the treatment one or more of: pain, chronic pain, acute pain, cramps, migraine, convulsions, stress, distress, anxiety, nausea, chemotherapy-induced nausea, vomiting, AIDS-related weight loss, wasting, spasm, multiple sclerosis, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, glaucoma, epilepsy, cancer.

19. A method of treating one or more of pain, chronic pain, acute pain, cramps, migraine, convulsions, stress, distress, anxiety, nausea, chemotherapy-induced nausea, vomiting, AIDS-related weight loss, wasting, spasm, multiple sclerosis, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, glaucoma, epilepsy and cancer, said method comprising administering the composition of claim 16 to a subject in need thereof.

20.

A method, use or composition substantially as described herein and with reference to the description or Examples.

21 01 19

Amendments to the claims have been filed as follows:

1. Use of ozone in a method for converting a non-psychoactive cannabinoid to a psychoactive cannabinoid, said use comprising providing a source of nonpsychoactive cannabinoid, contacting the source of non-psychoactive cannabinoid with ozone and optionally isolating the psychoactive cannabinoid.