Method of continuously producing cannabinol from cannabis plant and uses thereof

Abstract

Provided are: a method of preparing a cannabis processed product having an increased CBN content in an efficient and economic manner, through cyclization of CBD and aromatization of THC by continuous microwave irradiation of a cannabis extract; and use of a processed product having an increased CBN content prepared by the method, a fraction thereof, and a single ingredient of CBN, in foods, drugs, and cosmetics.

Claims

1. A method of producing cannabinoids, the method comprising irradiating microwaves to a reaction mixture comprising a Cannabis sp. plant or an extract thereof, an acid, and a solvent in a reaction vessel, wherein the microwave irradiation is carried out while flowing the reaction mixture from an inlet of the reaction vessel and out through an outlet of the reaction vessel, wherein the Cannabis sp. plant or the extract thereof comprises one or more of CBD and THC, and the cannabinoid is one or more of THC and CBN, wherein the reaction vessel is connected to a temperature control chamber for controlling the temperature inside the reaction vessel, wherein the reaction vessel is contained in the temperature control chamber filled with a liquid to control a temperature of the reaction mixture, wherein the temperature control chamber comprises a microwave-transparent material, and wherein the acid is methanesulfonic acid (MSA), benzenesulfonic acid, naphthalenesulfonic acid, toluenesulfonic acid, para-toluenesulfonic acid (p-toluensulfonic acid, PTSA), camphor-10-sulfonic acid (CSA), or a mixture thereof.

2. The method of claim 1, further comprising isolating cannabinoids from the microwave-irradiated reaction mixture.

3. The method of claim 1, wherein the extract is obtained by a method comprising contacting the Cannabis sp. plant with one or more of water, a protonic solvent, an aprotonic solvent, and a mixture thereof.

4. The method of claim 1, wherein the Cannabis sp. plant comprises leaves, flower buds, seeds, nuts, trichomes, flower bracts, stems, or any part comprising cannabinoids.

5. The method of claim 1, wherein the content of one or more of CBD and THC in the extract is 1% by weight or more, based on the total weight of the extract.

6. The method of claim 1, wherein the microwave irradiation is carried out at 60° C. to 150° C.

7. The method of claim 1, wherein the microwave irradiation is carried out for a time sufficient to convert one or more of CBD and THC into one or more of THC and CBN.

8. The method of claim 1, wherein the microwave irradiation is carried out for about 5 minutes to about 180 minutes in a continuous reactor.

9. The method of claim 1, wherein the microwave irradiation is carried out under pressure.

10. The method of claim 1, wherein the microwave irradiation is carried out at a pressure of 2 atm to 100 atm.

11. The method of claim 1, wherein the microwave irradiation is carried out at a frequency of 300 MHz to 300 GHz.

12. The method of claim 1, wherein the microwave irradiation is carried out at a power of 3 W to 6 kW.

13. The method of claim 1, wherein, in the microwave irradiation, the solvent is water, C1-C12 alcohol, or an aqueous solution thereof.

14. The method of claim 1, wherein the solvent is ethanol, isopropanol, butanol, or a 50% to 99% ethanol aqueous solution.

15. The method of claim 2, wherein the isolated cannabinoids comprise 5% by weight to 100% by weight of CBN, based on the total weight of the isolate.

16. The method of claim 1, wherein the reaction vessel comprises a tube between the inlet of the vessel and the outlet of the vessel.

17. The method of claim 1, wherein the reaction vessel is made of a microwave-transparent material.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The above and other aspects, features, and advantages of certain embodiments of the disclosure will be more apparent from the following description taken in conjunction with the accompanying drawings, in which:

(2) FIG. 1 shows an illustration of continuous microwave processing equipment;

(3) FIG. 2 shows a calibration curve constructed by analyzing CBD according to concentrations;

(4) FIG. 3 shows a calibration curve constructed by analyzing Δ.sup.9-THC according to concentrations;

(5) FIG. 4 shows a calibration curve constructed by analyzing CBN according to concentrations;

(6) FIG. 5 shows a UPLC chromatogram for analyzing cannabinoid ingredients in an extract A of a raw material dry cannabis leaf;

(7) FIG. 6 shows a UPLC chromatogram for analyzing cannabinoid ingredients in an extract B of a raw material dry cannabis leaf;

(8) FIG. 7 shows a UPLC chromatogram for analyzing cannabinoid ingredients in an extract C of a raw material dry cannabis leaf;

(9) FIG. 8 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation in a batch manner at 80° C. for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol;

(10) FIG. 9 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation in a batch manner at 80° C. for 60 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol;

(11) FIG. 10 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 40 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 60° C. at a flow rate of 0.25 mL/min;

(12) FIG. 11 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 60 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 60° C. at a flow rate of 0.017 mL/min;

(13) FIG. 12 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 40 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 70° C. at a flow rate of 0.025 mL/min;

(14) FIG. 13 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 60 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 70° C. at a flow rate of 0.017 mL/min;

(15) FIG. 14 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.050 mL/min;

(16) FIG. 15 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 90° C. at a flow rate of 0.050 mL/min;

(17) FIG. 16 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.050 mL/min;

(18) FIG. 17 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 110° C. at a flow rate of 0.050 mL/min;

(19) FIG. 18 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 120° C. at a flow rate of 0.050 mL/min;

(20) FIG. 19 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(21) FIG. 20 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 40 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.025 mL/min;

(22) FIG. 21 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in isopropanol (IPA) and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(23) FIG. 22 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethyl acetate (EtOAc) and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(24) FIG. 23 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in butanol (BuOH) and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(25) FIG. 24 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in a 70% ethanol (EtOH) aqueous solution and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(26) FIG. 25 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of sulfuric acid, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(27) FIG. 26 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of camphorsulfonic acid (CSA), relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(28) FIG. 27 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of methanesulfonic acid (MSA), relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(29) FIG. 28 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 10 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(30) FIG. 29 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 30 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(31) FIG. 30 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 40 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min;

(32) FIG. 31 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 10 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, to a 200 ppm solution of the extract B of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.10 mL/min;

(33) FIG. 32 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 10 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, to a 200 ppm solution of the extract B of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.050 mL/min;

(34) FIG. 33 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, to a 200 ppm solution of the extract B of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.033 mL/min;

(35) FIG. 34 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 10 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, to a 200 ppm solution of the extract C of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.100 mL/min;

(36) FIG. 35 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, to a 200 ppm solution of the extract C of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.050 mL/min; and

(37) FIG. 36 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, to a 200 ppm solution of the extract C of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.033 mL/min.

DETAILED DESCRIPTION

(38) Reference will now be made in detail to embodiments, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to like elements throughout. In this regard, the present embodiments may have different forms and should not be construed as being limited to the descriptions set forth herein. Accordingly, the embodiments are merely described below, by referring to the figures, to explain aspects of the present description. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items. Expressions such as “at least one of,” when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

(39) Hereinafter, the present disclosure will be described in more detail with reference to exemplary embodiments. However, these exemplary embodiments are only for illustrating the present disclosure, and the scope of the present disclosure is not limited to these exemplary embodiments.

Example 1: Preparation of CBN by Continuous Microwave Irradiation to Cannabis Extract

(40) In the following exemplary embodiments, cannabis was incubated in a solvent to obtain a cannabis extract, which was then continuously irradiated with microwaves to efficiently synthesize CBN.

(41) 1. Preparation of Cannabis Extract

(42) (1) Preparation of Cannabis Ethyl Acetate Extract from Korean Hemp (Including Only CBD)

(43) The Korean Hemp was provided by JayHempKorea Ltd., located in Sangju city, Gyeongsangbuk-do, South Korea, through assignment/transfer approval processes under drug (cannabis) research permission (No. 1564) obtained from the Ministry of Food and Drug Safety and Seoul Regional Food and Drug Administration. Cannabis seed shells, cannabis leaves, cannabis stems, and cannabis roots were harvested in October, 2018, and used. Cannabis leaves having a relatively high content of cannabinoids among the parts of cannabis were dried using a drying oven (Hanbaek Science Co. Ltd., HB-502L) at 80° C. for 48 hours, and then finely cut. 5 g of the dried cannabis leaves and 50 mL of ethyl acetate were added in a 250 mL Erlenmeyer flask, and microwave-irradiated using an ultrasonic processor (Sonics, VC505) at 40% power of the instrument for 1 hour, i.e., 200 W, and then incubated at room temperature for 24 hours. This procedure was repeated twice.

(44) The liquid extract, which was obtained by filtering the microwave-irradiated mixture of the dried cannabis leaves and ethyl acetate through a filter, was concentrated by evaporation under reduced pressure to obtain 0.44 g of a dry cannabis leave extract including CBD. Components of this extract were analyzed using UPLC-MS instruments (SHIMADZU), and as a result, Δ.sup.9-THC was not detected. Hereinafter, this extract was referred to as extract A.

(45) (2) Preparation of Cannabis Ethyl Acetate Extract from Cannabis (Including CBD and Δ.sup.9-THC)

(46) The Cannabis was provided by Andongpo Association, Inc., located in Andong city, Gyeongsangbuk-do, South Korea, through assignment/transfer approval processes under drug (cannabis) research permission (No. 1564) obtained from the Ministry of Food and Drug Safety and Seoul Regional Food and Drug Administration. Cannabis leaves were harvested in July, 2019, and dried using a drying oven (Hanbaek Science Co. Ltd., HB-502L) at 80° C. for 48 hours, and then finely cut and used. 5 g of the dried cannabis leaves and 50 mL of ethyl acetate were added in a 250 mL Erlenmeyer flask, and microwave-irradiated using an ultrasonic processor (Sonics, VC505) at 40% power of the instrument for 1 hour, i.e., 200 W, and then incubated at room temperature for 24 hours. This procedure was repeated twice.

(47) The liquid extract, which was obtained by filtering the microwave-irradiated mixture of the dried cannabis leaves and ethyl acetate through a filter, was concentrated by evaporation under reduced pressure to obtain 0.41 g of a dry cannabis leave extract including CBD and Δ.sup.9-THC. Components of this extract were analyzed using UPLC-MS instruments (SHIMADZU), and as a result, CBD and Δ.sup.9-THC were detected. Hereinafter, this extract was referred to as extract B.

(48) (3) Preparation of Cannabis Ethyl Acetate Extract from Indigenous Species of Cannabis (Including Only Δ.sup.9-THC)

(49) An indigenous species of cannabis was provided by Andongpo Association, Inc., located in Andong city, Gyeongsangbuk-do, South Korea, through assignment/transfer approval processes under drug (cannabis) research permission (No. 1564) obtained from the Ministry of Food and Drug Safety and Seoul Regional Food and Drug Administration. Cannabis leaves were harvested in July, 2019, and dried using a drying oven (Hanbaek Science Co. Ltd., HB-502L) at 80° C. for 48 hours, and then finely cut and used. 5 g of the dried cannabis leaves and 50 mL of ethyl acetate were added in a 250 mL Erlenmeyer flask, and microwave-irradiated using an ultrasonic processor (Sonics, VC505) at 40% power of the instrument for 1 hour, i.e., 200 W, and then incubated at room temperature for 24 hours. This procedure was repeated twice.

(50) The liquid extract, which was obtained by filtering the microwave-irradiated mixture of the dried cannabis leaves and ethyl acetate through a filter, was concentrated by evaporation under reduced pressure to obtain 0.25 g of a dry cannabis leave extract including Δ.sup.9-THC. Components of this extract were analyzed using UPLC-MS instruments (SHIMADZU), and as a result, only Δ.sup.9-THC was detected. Hereinafter, this extract was referred to as extract C.

(51) 2. Manufacture of Continuous Microwave Processing Equipment

(52) As a continuous microwave processing equipment, a microwave irradiator (model no. 908005) manufactured by CEM Company (USA) was used, and a tube made of PTFE and PFA was inserted into a reaction chamber of a 10-mL flow cell accessory (model no. 908910) to manufacture a continuous reactor. Thereafter, the chamber of the continuous reactor was filled with water, and a liquid-feeding pump (YMC-KP series) was connected to one end of the inserted tube, i.e., to a tube at an inlet through which the reaction mixture is applied, and a back pressure regulator of 75 psi (UPCHURCH, P-786) was connected to the other end, i.e., to a tube at an outlet through which the reaction mixture is discharged. The tube had an outer diameter (O.D.) of 1/16 inch, an inner diameter (I.D.) of 1.0 mm, a length of 127.4 cm, and an inner tube volume of 1.0 mL.

(53) FIG. 1 shows an illustration of continuous microwave processing equipment.

(54) 3. Continuous Microwave Processing of Cannabis Leaf Extracts A and B

(55) (1) Experimental Group

(56) The cannabis leaf extracts A and B were subjected to continuous microwave processing. In detail, the cannabis leaf extract A was dissolved at a concentration of 200 ppm in ethanol, and then 20 equivalent weights of p-toluenesulfonic acid (PTSA), relative to CBD, was added thereto. A reaction temperature of the continuous reactor was set at 60° C., and continuous microwave processing was carried out at a microwave maximum power of 50 W and a frequency of 2450 MHz for 40 minutes (experimental group 1) and 60 minutes (experimental group 2). In such a manner, experiments were performed at 70° C. for 40 min (experimental group 3) and at 70° C. 60 min (experimental group 4), at 80° C. for 20 min (experimental group 5), at 90° C. for 20 min (experimental group 6), at 100° C. for 20 min (experimental group 7), at 110° C. for 20 min (experimental group 8), and at 120° C. for 20 min (experimental group 9). Thereafter, a reaction temperature of the continuous reactor was set at 100° C., and microwave irradiation was performed for 30 min (experimental group 10) and 40 minutes (experimental group 11) to obtain microwave-irradiated processed products, respectively.

(57) Next, the solvent was replaced by isopropanol (experimental group 12), ethyl acetate (experimental group 13), butanol (experimental group 14), and 70% ethanol aqueous solution (experimental group 15), and microwave irradiation was performed to obtain processed products, respectively.

(58) Next, the reaction temperature of the continuous reactor was set at 100° C., and 20 equivalent weights of sulfuric acid (experimental group 16), camphorsulfonic acid (CSA) (experimental group 17), or methanesulfonic acid (MSA) (experimental group 18), relative to CBD, was added for 30 minutes to perform experiments, and 10 equivalent weights (experimental group 19), 30 equivalent weights (experimental group 20), or 40 equivalent weights (experimental group 21) of PTSA was added, and each was irradiated with microwaves to obtain processed products, respectively.

(59) Next, the cannabis leaf extract B was dissolved at a concentration of 200 ppm in ethanol, and then 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, was added thereto. A reaction temperature of the continuous reactor was set at 80° C., and continuous microwave processing was carried out at a microwave maximum power of 50 W and a frequency of 2450 MHz for 10 minutes (experimental group 22), 20 minutes (experimental group 23) and 30 minutes (experimental group 24) to obtain processed products, respectively.

(60) Next, the cannabis leaf extract C was dissolved at a concentration of 200 ppm in ethanol, and then 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, was added thereto. A reaction temperature of the continuous reactor was set at 80° C., and continuous microwave processing was carried out at a microwave maximum power of 50 W and a frequency of 2450 MHz for 10 minutes (experimental group 25), 20 minutes (experimental group 26) and 30 minutes (experimental group 27) to obtain processed products, respectively.

(61) Each reaction time was controlled by controlling the flow rate of the liquid-feeding pump, and the power was 3 W to 50 W during microwave irradiation, and the content analysis was performed according to an analysis method of the following section 4. The reaction time according to the flow rate was as follows: 5 min at 0.2 mL/min, 10 min at 0.1 mL/min, 20 min at 0.05 mL/min and 30 min at 0.033 mL/min, 40 min at 0.025 mL/min, and 60 min at 0.017 mL/min. Here, the reaction time represents the time for which the reactants remain in the tube in the continuous reactor. During the microwave irradiation, the power may vary depending on the size of the inner diameter of the tube.

(62) (2) Control Group

(63) For experiments of control groups, the cannabis leaf extract A was irradiated with microwaves in a batch manner, instead of the continuous manner. In detail, the extract A of dry cannabis leaves was dissolved at a concentration of 200 ppm in ethanol, and then 20 equivalent weights of PTSA, relative to CBD, was added thereto. A reaction temperature of a batch-type reactor was set at 80° C., and heat treatment was carried out for 30 minutes (control group 1) and 60 minutes (control group 2) to obtain processed products, respectively.

(64) 4. Analysis of Cannabinoids in Extracts and Continuous or Batch-Type Microwave-Processed Products

(65) (1) Experimental Method

(66) Based on values of CBD, Δ.sup.9-THC, and CBN calibration curves, cannabinoids in the cannabis extracts A, B, and C, and the processed extracts obtained by batch-type or continuous microwave irradiation thereof were analyzed, and repeated in triplicate to confirm reproducibility. As for CBD, Δ.sup.9-THC, and CBN single ingredients used in the experiments, CBD with purity of 96.3%, Δ.sup.9-THC with purity of 96.8%, and CBN with purity of 96.7% directly isolated from the cannabis leaf raw material were used. According to the general calibration curve analysis method, CBD, Δ.sup.9-THC, and CBN were dissolved in water at 10 ppm, 25 ppm, 50 ppm, 100 ppm, and 250 ppm to prepare standard solutions, respectively, which were used to construct calibration curves. An elution solvent A and an elution solvent B used in ultra-performance liquid chromatography (UPLC) were water and acetonitrile, respectively, and each was pumped using two pumps. 3 μl of the standard aqueous solution was injected into a reverse-phase column for analysis (Phenomenex Luna Omega 1.6μ Polar C18, 150 mm×2.1 mm) using a syringe, and an elution solvent consisting of 70% by volume of A and 30% by volume of B was applied at a flow rate of 0.3 mL/min. Thereafter, % volume of the elution solvent B were gradually changed to 100% (20 min), 100% (23 min), and 30% (26 min). After the above procedures, each ingredient isolated from the column was analyzed by UV spectrum.

(67) (2) Experimental Results

(68) As a result of the experiments, each ingredient isolated from the column was analyzed by UPLC analysis of the cannabis leaf extracts, and peaks of FIGS. 2 to 36 were obtained by the analysis results of UPLC chromatograms.

(69) FIG. 2 shows a calibration curve constructed by analyzing CBD according to concentrations.

(70) FIG. 3 shows a calibration curve constructed by analyzing Δ.sup.9-THC according to concentrations.

(71) FIG. 4 shows a calibration curve constructed by analyzing CBN according to concentrations.

(72) FIG. 5 shows a UPLC chromatogram for analyzing cannabinoid ingredients in an extract A of a raw material dry cannabis leaf.

(73) FIG. 6 shows a UPLC chromatogram for analyzing cannabinoid ingredients in an extract B of a raw material dry cannabis leaf.

(74) FIG. 7 shows a UPLC chromatogram for analyzing cannabinoid ingredients in an extract C of a raw material dry cannabis leaf.

(75) FIG. 8 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation in a batch manner at 80° C. for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol.

(76) FIG. 9 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation in a batch manner at 80° C. for 60 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol.

(77) FIG. 10 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 40 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 60° C. at a flow rate of 0.25 mL/min.

(78) FIG. 11 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 60 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 60° C. at a flow rate of 0.017 mL/min.

(79) FIG. 12 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 40 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 70° C. at a flow rate of 0.025 mL/min.

(80) FIG. 13 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 60 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 70° C. at a flow rate of 0.017 mL/min.

(81) FIG. 14 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.050 mL/min.

(82) FIG. 15 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 90° C. at a flow rate of 0.050 mL/min.

(83) FIG. 16 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.050 mL/min.

(84) FIG. 17 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 110° C. at a flow rate of 0.050 mL/min.

(85) FIG. 18 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 120° C. at a flow rate of 0.050 mL/min.

(86) FIG. 19 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(87) FIG. 20 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 40 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.025 mL/min.

(88) FIG. 21 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in isopropanol (IPA) and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(89) FIG. 22 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethyl acetate (EtOAc) and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(90) FIG. 23 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in butanol (BuOH) and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(91) FIG. 24 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in a 70% ethanol (EtOH) aqueous solution and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(92) FIG. 25 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of sulfuric acid, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(93) FIG. 26 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of camphorsulfonic acid (CSA), relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(94) FIG. 27 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of methanesulfonic acid (MSA), relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(95) FIG. 28 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 10 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(96) FIG. 29 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 30 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(97) FIG. 30 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 40 equivalent weights of PTSA, relative to CBD, to a 200 ppm solution of the extract A of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 100° C. at a flow rate of 0.033 mL/min.

(98) FIG. 31 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 10 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, to a 200 ppm solution of the extract B of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.10 mL/min.

(99) FIG. 32 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 10 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, to a 200 ppm solution of the extract B of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.050 mL/min.

(100) FIG. 33 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of CBD and Δ.sup.9-THC, to a 200 ppm solution of the extract B of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.033 mL/min.

(101) FIG. 34 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 10 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, to a 200 ppm solution of the extract C of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.100 mL/min.

(102) FIG. 35 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 20 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, to a 200 ppm solution of the extract C of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.050 mL/min.

(103) FIG. 36 shows a UPLC chromatogram for analyzing cannabinoid ingredients in a processed product obtained by microwave-irradiation for 30 minutes while adding 20 equivalent weights of PTSA, relative to the total weight of Δ.sup.9-THC, to a 200 ppm solution of the extract C of cannabis leaves in ethanol and applying the solution to a tube with a volume of 1.0 mL through a liquid-feeding pump at 80° C. at a flow rate of 0.033 mL/min.

(104) First, the results of calculating the contents of CBD, Δ.sup.9-THC, and CBN in UPLC chromatograms obtained by continuous microwave processing of the cannabis extracts at different temperatures for different times are summarized in Table 1.

(105) TABLE-US-00001 TABLE 1 Tem- Acid- perature equiv- (° C.)- alent Δ.sup.9- CBN time weight CBD THC CBN yield* Item (min) (eq) (mg) (mg) (mg) (%) Extract A — 62.2 n.d n.d 0 Control group 1 80-30 PTSA-20 n.d 39.2 n.d 0 Control group 2 80-60 PTSA-20 n.d 34.4 n.d 0 Experimental 60-40 PTSA-20 12.0 27.8 4.2 6.8 group 1 Experimental 60-60 PTSA-20 4.4 25.4 8.7 14.2 group 2 Experimental 70-40 PTSA-20 n.d 14.6 25.0 40.7 group 3 Experimental 70-60 PTSA-20 n.d 10.0 30.2 49.2 group 4 Experimental 80-20 PTSA-20 n.d 22.0 26.3 42.8 group 5 Experimental 90-20 PTSA-20 n.d 15.3 33.4 54.4 group 6 Experimental 100-20  PTSA-20 n.d 16.0 35.9 58.5 group 7 Experimental 110-20  PTSA-20 n.d 18.4 6.6 10.7 group 8 Experimental 120-20  PTSA-20 n.d 9.4 3.3 5.4 group 9 Experimental 100-30  PTSA-20 n.d 15.2 37.1 60.4 group 10 Experimental 100-40  PTSA-20 n.d 11.4 36.0 58.6 group 11
*CBN yield=(CBN mg/61.4 mg(Theoretical CBN amount at 100% conversion)×100**n.d=not detected

(106) Table 1 shows CBD, Δ.sup.9-THC, and CBN contents expressed in mg per 1 g of the extract, after dissolving the extract A of dry cannabis leaves in ethanol at a concentration of 200 ppm, adding 20 equivalent weights of PTSA, relative to CBD, and processing by continuous microwave irradiation. It was confirmed that Δ.sup.8-THC and cannabicitran were produced as by-products during processing. However, they were inseparable mixtures, and thus the exact production amounts thereof were not calculated. In the initial cannabis leaf extract A during processing, cannabinoids were found to include only 62.2 mg of CBD per g, and Δ.sup.9-THC and CBN were not found.

(107) Prior to this experiment, in order to compare the batch method and the flow method, an experiment was performed using a batch-type microwave reactor at 80° C. for 30 minutes (control group 1) and 60 minutes (control group 2). As a result, the production of Δ.sup.9-THC through cyclization of CBN was found, but the conversion to CBN through aromatization was not found. However, when reaction was allowed for 40 minutes at 60° C. in the continuous manner, it was confirmed that CBD remained, but the final product CBN was produced via the intermediate Δ.sup.9-THC. Thereafter, experiments were performed at 60° C. to 120° C. at 10° C. intervals, and measurements were performed between 10 minutes and 60 minutes to examine the influence of time on temperature. First, when the experiment was performed for up to 60 minutes at 60° C., the raw material CBD remained, and the largest amount of the intermediate Δ.sup.9-THC was observed. When the temperature was raised to 70° C., CBD was exhausted and CBN was increased. Next, the time was fixed at 20 minutes, and the experiment was performed while raising the temperature from 80° C. to 120° C. As a result, the best result was obtained at 100° C. and 20 minutes. Finally, it was confirmed that when the experiment was performed at 100° C. by increasing the time, CBD was converted to CBN with the highest yield of 60.4% in 30 minutes.

(108) Further, the results of calculating the contents of CBD, Δ.sup.9-THC, and CBN in UPLC chromatograms obtained by continuous microwave processing of the cannabis extracts in different solvents are summarized in Table 2.

(109) TABLE-US-00002 TABLE 2 Tem- Acid- perature equiv- (° C.)- alent Δ.sup.9- CBN Sol- time weight CBD THC CBN yield* Item vent (min) (eq) (mg) (mg) (mg) (%) Extract A — 62.2 0 0   0% Experi- EtOH 100-30 PTSA- n.d 15.2 37.1 60.4% mental 20 group 10 Experi- IPA 100-30 PTSA- n.d 14.9 16.5 26.9% mental 20 group 11 Experi- EtOAc 100-30 PTSA- n.d 9.8 23.5 38.3% mental 20 group 12 Experi- BuOH 100-30 PTSA- n.d 15.8 28.3 46.1% mental 20 group 13 Experi- 70% 100-30 PTSA- n.d 17.0 31.2 50.8% mental EtOH 20 group 14
*CBN yield=(CBN mg/61.4 mg(Theoretical CBN amount at 100% conversion)×100**n.d=not detected

(110) Table 2 shows CBD, Δ.sup.9-THC, and CBN contents expressed in mg per 1 g of the extract, after dissolving the extract A of dry cannabis leaves in each solvent at a concentration of 200 ppm, adding 20 equivalent weights of PTSA, relative to CBD, and processing by continuous microwave irradiation under conditions of experimental group 10 (100° C., 30 minutes) showing the highest CBN yield in Table 1. As a result, when the solvent isopropanol (IPA), ethyl acetate (EtOAc), butanol (BuOH), and 70% EtOH aqueous solution were used, the CBN content was 26.9%, 38.3%, 46.1%, and 50.8%, respectively. The yield was lower than that of ethanol, but CBN was obtained.

(111) Further, the results of calculating the contents of CBD, Δ.sup.9-THC, and CBN in UPLC chromatograms obtained by continuous microwave processing of the cannabis extracts with different acids and equivalent weights are summarized in Table 3.

(112) TABLE-US-00003 TABLE 3 Temperature Acid- CBN (° C.)-time equivalent CBD Δ.sup.9-THC CBN yield* Item (min) weight (eq) (mg) (mg) (mg) (%) Extract A 62.2 0 0 0 Experimental 100-30 PTSA-20 n.d 15.2 37.1 60.4 group 10 Experimental 100-30 Sulfuric n.d 7.6 4.8 7.8 group 16 acid-20 Experimental 100-30 CSA-20 n.d 9.1 32.2 52.4 group 17 Experimental 100-30 MSA-20 n.d 20.0 12.3 20.0 group 18 Experimental 100-30 PTSA-10 n.d 5.3 32.1 52.3 group 19 Experimental 100-30 PTSA-30 n.d 15.5 36.5 59.4 group 20 Experimental 100-30 PTSA-40 n.d 16.3 37.2 60.6 group 21
*CBN yield=(CBN mg/61.4 mg(Theoretical CBN amount at 100% conversion)×100**n.d=not detected

(113) Table 3 shows CBD, Δ.sup.9-THC, and CBN contents expressed in mg per 1 g of the extract, after dissolving the extract A of dry cannabis leaves in each solvent at a concentration of 200 ppm, and processing by continuous microwave irradiation with varying the kind of acid and the equivalent weight under conditions of experimental group 10 (100° C., 30 minutes) showing the highest CBN yield in Tables 1 and 2. As a result, when sulfuric acid, camphorsulfonic acid (CSA), and methanesulfonic acid (MSA) were used, the CBN content was 7.8%, 52.4%, and 20.0%, respectively. The yield was lower than that of PTSA, but CBN was produced. According to the results, when the same experiment was performed by varying the equivalent weight of PTSA showing the highest yield to 10 equivalent weights, 30 equivalent weights, and 40 equivalent weights, the yield was decreased to 52.3% when 10 equivalent weights of PTSA was used, but there was little change when 20 equivalent weights to 40 equivalent weights of PTSA were used.

(114) Further, the results of calculating the contents of CBD, Δ.sup.9-THC, and CBN in UPLC chromatograms obtained by continuous microwave processing of the cannabis extract B including both CBD and Δ.sup.9-THC are summarized in Table 4.

(115) TABLE-US-00004 TABLE 4 Temperature Acid- CBN (° C.)-time equivalent CBD Δ.sup.9-THC CBN yield* Item (min) weight (eq) (mg) (mg) (mg) (%) Extract B — 52.3 84.9 n.d 0 Experimental 80-10 PTSA-20 n.d 14.8 96.6 71.3 group 22 Experimental 80-20 PTSA-20 n.d 7.7 98.1 72.4 group 23 Experimental 80-30 PTSA-20 n.d 6.8 93.4 69.0 group 24
*CBN yield=(CBN mg/135.44 mg(Theoretical CBN amount at 100% conversion)×100**n.d=not detected

(116) Table 4 shows CBD, Δ.sup.9-THC, and CBN contents expressed in mg per 1 g of the extract, after dissolving the extract B of dry cannabis leaves in ethanol at a concentration of 200 ppm, adding 20 equivalent weights of PTSA, relative to CBD, and processing by continuous microwave irradiation at 80° C. for 10 minutes, 20 minutes, and 30 minutes. As a result, conversion of CBD and Δ.sup.9-THC to CBN was 71.3%, 72.4% and 69.0%, respectively.

(117) Further, the results of calculating the contents of CBD, Δ.sup.9-THC, and CBN in UPLC chromatograms obtained by continuous microwave processing of the cannabis extract C including only Δ.sup.9-THC are summarized in Table 5.

(118) TABLE-US-00005 TABLE 5 Temperature Acid- CBN (° C.)-time equivalent CBD Δ.sup.9-THC CBN yield* Item (min) weight (eq) (mg) (mg) (mg) (%) Extract C — n.d 71.1 mg n.d   0% Experimental 80-10 PTSA-20 n.d 15.7 mg 50.9 mg 71.5% group 25 Experimental 80-20 PTSA-20 n.d  5.3 mg 61.2 mg 86.0% group 26 Experimental 80-30 PTSA-20 n.d  3.0 mg 58.2 mg 81.9% group 27
*CBN yield=(CBN mg/135.44 mg(Theoretical CBN amount at 100% conversion)×100**n.d=not detected

(119) Table 5 shows CBD, Δ.sup.9-THC, and CBN contents expressed in mg per 1 g of the extract, after dissolving the extract C in ethanol at a concentration of 200 ppm, adding 20 equivalent weights of PTSA, relative to Δ.sup.9-THC, and processing by continuous microwave irradiation at 80° C. for 10 minutes, 20 minutes, and 30 minutes. As a result, as shown in Table 5, conversion of Δ.sup.9-THC to CBN was 71.5%, 86.0%, and 81.9%, respectively.

(120) Therefore, according to the above method, only Δ.sup.9-THC was produced through the cyclization reaction of CBD under the batch-type microwave conditions, but production of CBN through aromatization was not found. Meanwhile, it was found that CBD was converted to CBN via Δ.sup.9-THC under the continuous microwave conditions, and thus a method capable of mass-producing a processed product with a high content of CBN was developed.

(121) As a result of the above experiments, when the cannabis leaf extracts A, B, and C were processed by the continuous microwave method, CBD or Δ.sup.9-THC which is the main cannabinoid component of cannabis, were efficiently converted into CBN. In detail, it was possible to obtain a continuous microwave-processed product, in which the major cannabinoid CBD and Δ9-THC complex components of cannabis were converted to the trace cannabinoid CBN with a conversion rate of 5.4% to 86.0%.

(122) According to the method of producing cannabinoids according to an aspect, one or more of THC and CBN may be efficiently produced.

(123) The composition according to another aspect may be used for anti-epilepsy, neuroprotection, vasorelaxation, anti-cancer, anti-inflammation, anti-diabetes, anti-bacteria, analgesia, anti-osteoporosis, immune enhancement, or antiemetic action, health functional foods, or cosmetics.

(124) It should be understood that embodiments described herein should be considered in a descriptive sense only and not for purposes of limitation. Descriptions of features or aspects within each embodiment should typically be considered as available for other similar features or aspects in other embodiments. While one or more embodiments have been described with reference to the figures, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the disclosure as defined by the following claims.