DESCRIPTION

FIELD OF THE INVENTION

The present invention relates to a method for the preparation of cannabis oil. In particular, the present invention relates to a highly efficient method for the preparation of cannabis oil with a high concentration of delta-9-tetrahydrocannabinol (“THC”) and cannabidiol (“CBD”), as well as other substances of therapeutic interest contained therein.

STATE OF THE ART The term cannabis is used to refer to the plant Cannabis sativa L, selected over the years for the large quantities of active ingredients of therapeutic interest contained therein.

In cannabis, the active ingredients of therapeutic interest are contained in the unfertilized female inflorescences, very rich in glandular structures called trichomes. In cannabis, the two most representative active ingredients are delta-9- tetrahydrocannabinol (“THC”) and cannabidiol (“CBD”), contained in the inflorescences in the form of prodrugs in acid form, inactive or with very little activity under the therapeutic profile, respectively delta-9-tetrahydrocannabinolic acid (“THCA”) and cannabidiolic acid (“CBDA”). Although the presence of these active ingredients in cannabis has been known and exploited for recreational purposes for a very long time, only for some years has cannabis attracted scientific interest for the possibility of exploiting the properties of these active ingredients contained therein for therapeutic purposes.

Unlike conventional drug studies, where receptor models and molecules which have an affinity or rather the highest possible receptor specificity for that type of receptor are studied, the study of cannabis has instead been “retrospective”, i.e. , the scientific community has begun to study the possible mechanisms of action of the active ingredients of cannabis, only following the dissemination of information and data about the therapeutic effects attributable to this plant. In this context, the most important discovery concerned the endocannabinoid system, which, like that already known for endorphins, explains how the cannabinoids produced by the body, through the interaction with the respective receptors, are responsible for causing the sensation of relaxation and the resulting well-being.

Even at the legislative level, countries have only recently regulated the possibilities of prescribing cannabis and/or the derivatives thereof for therapeutic purposes. For example, Italy passed Law no. 94 of 8/4/1998 (“Di Bella Law”) which allows each doctor to prescribe cannabis for therapeutic purposes, and has instructed the Florentine Pharmaceutical Institute to start the production of a domestic cannabis which was previously exclusively imported from abroad.

The active ingredients of cannabis are generally used in the form of an oily matrix extract, also known as “cannabis oil”. Numerous techniques on how to obtain a cannabis oil are known, which generally involve a combination of extraction and decarboxylation steps of the prodrugs in acid form, to obtain an oil containing active ingredients of therapeutic interest.

For example, in Cannabinoids Vol. 7, Issue 1 of 5 May 2013; 1 (1 ): 1-11 , L. Romano and A. Hazekamp describe a prodrug extraction and decarboxylation technique (“Romano Hazekamp” method) which involves a pre-grinding of the cannabis inflorescences and a step of hot extraction of the cannabis inflorescences by means of olive oil Ph. Eur., in which the ratio between the inflorescence and the solvent is 1 :10 (5-10 g of cannabis in 50-100 ml of olive oil). Subsequently, the oil containing the pre-ground inflorescences is heated under stirring to a temperature of 98°C in a water bath for 120 minutes. Furthermore, the “Oily extraction of female cannabis inflorescences” procedure proposed by the SIFAP working group (Italian Society of Preparatory Pharmacists) (link: https://www.sifap.org/procedure/estrazione-oieosa-di-infiore scenze-femminili- di-cannabi) includes preparing oily dispersions of cannabis inflorescences in O.P olive oil OP (Official Pharmacopoeia), starting from an inflorescence: oil ratio equal to 1 g: 10 ml. According to said method (“SIFAP method”), before being put in contact with the oil, the inflorescences are chopped, thermally pre-treated at a temperature of about 115°C for 40 minutes in a heater, and left to cool for at least 10 minutes, and lastly they are subsequently mixed with oil. The oily dispersion thus obtained is then heated to 100°C for about 40 minutes in a water bath.

Cannazza et al. (G. Cannazza et al. “Medicinal cannabis: Principal cannabinoids concentration and their stability evaluated by a high performance liquid chromatography coupled to diode array and quadrupole time of flight mass spectrometry method” Elsevier, Journal of Pharmaceutical and Biomedical Analysis 128(2016) 201-209) describes the preparation (“Cannazza” method) of a cannabis oil in which the finely pulverized inflorescences are dispersed in olive oil (10 ml of oil for each gram of inflorescences) and said dispersion is heated under stirring to 110°C under reflux for 2 hours. The sample is then gradually cooled in air and filtered through paper.

Finally, Calvi (L.Calvi et al. Comprehensive quality evaluation of medical Cannabis sativa L. inflorescence and macerated oils based on HS-SPME coupled to GC-MS and LC-HRMS (q-exactive orbitrap ®) approach” Elsevier, Journal of Pharmaceutical and Biomedical Analysis 150(2018) 208 -219) describes the preparation (“Calvi” method) of a cannabis oil in which the inflorescences are ground in a planetary ball mill with a frequency of 35 Hz for 1 minute in containers cooled with liquid nitrogen, and subsequently heated in a static oven at 145° for 30 minutes. The inflorescences thus treated are then placed inside amber glass bottles with oil (10 ml of oil for each gram of inflorescences) and the dispersion thus obtained is sonicated (35 kHz) for 30 minutes without further heating and subsequently filtered through paper.

The Applicant has perceived that the techniques of the known art for the preparation of cannabis oil have some performance and application limits.

The Applicant has in particular noted that the control of the conditions of the decarboxylation reaction of the prodrugs THCA and CBDA to create THC and CBD can significantly influence the quality of the cannabis oil.

In particular, after studying the criticalities present in the methods indicated above, the Applicant has developed a new method which, together with the original equipment object of the present invention, allows a better efficiency in the extraction of the active ingredients, of the decarboxylation process which affects the activation of the known cannabinoids THCA and CBDA in the corresponding neutral active ingredients THC and CBD and together with these a greater efficiency in the extraction of the entire phytocomplex which also contains the terpenes.

The Applicant has therefore found that there is a need for a new easy-to-use method for the preparation of cannabis oil, capable of obtaining a high concentration of THC and CBD and other active substances which contribute to the therapeutic effect, such as the non-cannabinoid active ingredients, for example the terpene compounds.

SUMMARY OF THE INVENTION

In accordance with the present invention, the Applicant has surprisingly found that these desired features can be obtained by means of a process which includes carrying out the decarboxylation reaction under specific conditions such as to allow obtaining high concentrations of THC and CBD, and at the same time to increase the content of other active substances which contribute to the therapeutic effect, such as the non-cannabinoid active ingredients, for example the terpene compounds, in the cannabis oil.

Therefore, the present invention relates in a first aspect to a method for the preparation of cannabis oil, comprising the steps of: a) providing at least one oily dispersion by dispersing at least one unfertilized female cannabis inflorescence in at least one vegetable oil; b) heating said oily dispersion to a temperature ranging from 90°C to 150°C and for a time ranging from 20 to 150 minutes, in at least one hermetically sealed reactor; c) bringing, in the at least one hermetically sealed reactor, the at least one oily dispersion obtained from step b) to a temperature ranging from 5°C to 35°C; and d) separating at least one liquid component from at least one solid component of the oily dispersion obtained from step c), so as to obtain the cannabis oil.

It has in fact surprisingly been discovered that by heating an oily dispersion of said at least one cannabis inflorescence in said at least one vegetable oil in a hermetically sealed reactor with a specific combination of temperatures and heating time, it is possible to decarboxylate the prodrugs present in acid form in said at least one inflorescence, and obtain a cannabis oil with a high concentration of THC and CBD and other active substances which contribute to the therapeutic effect, such as the non-cannabinoid active ingredients, for example the terpene compounds, of the cannabis plant.

The Applicant has also noted that the step of extraction in oil of the active substances contained in the unfertilized female cannabis inflorescences can significantly influence the qualitative and quantitative composition of the cannabis oil, in particular by limiting the content of THC and CBD and of other active ingredients which contribute to the therapeutic effect, such as the non-cannabinoid active ingredients, for example the terpene compounds, in the cannabis oil itself.

Therefore, in a preferred embodiment of the method according to the invention, in said step a) said at least one oily dispersion is stirred in at least one extractor provided with a stirring system, at a temperature ranging from 60°C to 120°C and at a pressure ranging from 2 bar to 40 bar, for a time ranging from 30 minutes to 150 minutes.

Indeed, it has surprisingly been discovered that, by subjecting the at least one oily dispersion to stirring in said specific combination of temperature, pressure and stirring time, it is possible to increase the titre in THC and CBD and other active substances which contribute to the therapeutic effect, in the cannabis oil.

In a preferred embodiment of the method according to the invention, said step b) of the method according to the invention is carried out by irradiating said at least one oily dispersion with a microwave electromagnetic radiation and said reactor of said step b) of the method according to the invention includes at least one wall comprising a part of a material transparent to microwave electromagnetic radiation.

In a further preferred embodiment of the method according to the invention, said step b) of the method according to the invention is carried out by heating said at least one oily dispersion by means of oscillating magnetic induction and said reactor of said step b) of the method according to the invention includes at least one wall of an electrically resistive material and/or provided with a magnetic hysteresis loop.

In a particularly preferred embodiment of the present invention, the reactor of said step b) of the method according to the invention is an apparatus comprising:

- a hollow body delimiting a seat adapted to contain said at least one oily dispersion, wherein said hollow body has at least one side wall delimiting an opening and a bottom wall which has at least one discharge channel;

- a piston slidably housed in said seat and adapted to hermetically slide against said at least one side wall and to hermetically close said seat;

- at least one fixing element housed at said opening and adapted to lock said piston in abutment and to prevent it from exiting from said seat;

- a filtering element housed in said seat at said bottom wall and adapted to separate said at least one oily dispersion into a solid component and a liquid component; and

- at least one discharge valve means fixed to said bottom wall at said discharge channel.

It has in fact been found that the use of such an apparatus is particularly advantageous for obtaining a cannabis oil with a high concentration of THC and CBD and other active substances deriving from the phytocomplex of the cannabis plant.

In fact, thanks to the possibility of using this apparatus, the Applicant has surprisingly discovered that it is possible to obtain the decarboxylation of the THCA and CBDA prodrugs of cannabis to create the therapeutically active forms THC and CBD, to obtain a high content of other active substances deriving from the phytocomplex of the cannabis plant, as well as to rapidly separate at least one liquid component from at least one solid component of the oily dispersion obtained from step d), so as to obtain the cannabis oil.

In a further aspect, the present invention relates to an apparatus comprising: - a hollow body delimiting a seat adapted to contain a dispersion of at least one solid component in at least one liquid component, wherein said hollow body has at least one side wall delimiting an opening and a bottom wall which has at least one discharge channel;

- a piston slidably housed in said seat and adapted to hermetically slide against said at least one side wall;

- at least one fixing element housed at said opening and adapted to lock said piston in abutment and to prevent it from exiting from said seat;

- a filtering element housed in said seat at said bottom wall and adapted to separate said dispersion into at least one solid component and at least one liquid component;

- at least one discharge valve means fixed to said bottom wall at said discharge channel; and

- at least one heating device adapted to heat a dispersion of at least one solid component in at least one liquid component contained in said seat, selected from a device adapted to generate a microwave electromagnetic radiation and a device adapted to generate an oscillating magnetic induction to produce heat exploiting Foucault eddy currents and/or a magnetic hysteresis loop; wherein said apparatus includes at least one wall comprising a part of a material selected from the group consisting of: material transparent to microwave electromagnetic radiation, and electrically resistive material and/or provided with a magnetic hysteresis loop.

Said apparatus is in fact particularly adapted to be used for use for the preparation of cannabis oil. The advantages of said apparatus have already been described with reference to the first aspect of the invention and are not repeated here.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows a schematic longitudinal-section view not to scale of an embodiment of an extractor adapted to be used in step a) of the method according to the present invention;

Figure 2 shows a schematic longitudinal-section view not to scale of an embodiment of an apparatus according to the present invention;

Figure 3 shows the chromatogram obtained in Example 2 - replication 2 through GC-MS using an Orbitrap qExtractive mass spectrometer;

Figure 4 shows the chromatogram obtained in Example 2 - replication 3 through GC-MS using an Orbitrap qExtractive mass spectrometer; and

Figure 5 shows a schematic longitudinal-section view not to scale of an embodiment of an extractor adapted to be used in step a) of the method according to the present invention, which also has a filtering and collection accessory.

DETAILED DESCRIPTION OF THE INVENTION

The present invention can be presented in one or more of the aspects thereof or one or more of the preferred features reported below, which can be combined with one another according to the application requirements.

Within the context of the present description and following claims, all the numerical magnitudes indicating quantities, parameters, percentages, and so on are to be considered preceded in every circumstance by the term “about” unless indicated otherwise. Further, all the ranges of numerical magnitudes include all the possible combinations of maximum and minimum numerical values and all the possible intermediate ranges, as well as those indicated below.

In the present invention when use is made of the expression:

“cannabis”, it means the plant Cannabis sativa L;

- “cannabis oil”, it means a solution in a vegetable oil of the active principles of the inflorescences of Cannabis, commonly also referred to as “oleolyte”;

“THC”, it means delta-9-tetrahydrocannabinol;

- “CBD”, it means cannabidiol;

- “THCA”, it means delta-9-tetrahydrocannabinolic acid; and “CBDA”, it means cannabidiolic acid.

In a first aspect thereof, the present invention relates to a method for the preparation of cannabis oil, comprising the steps of: a) providing at least one oily dispersion by dispersing at least one unfertilized female cannabis inflorescence in at least one vegetable oil; b) heating said oily dispersion to a temperature ranging from 90°C to 150°C and for a time ranging from 20 to 150 minutes, in at least one hermetically sealed reactor; c) bringing, in the at least one hermetically sealed reactor, the at least one oily dispersion obtained from step b) to a temperature ranging from 5°C to 35°C; and d) separating at least one liquid component from at least one solid component of the oily dispersion obtained from step c), so as to obtain the cannabis oil.

It has in fact surprisingly been discovered that by heating an oily dispersion of said at least one cannabis inflorescence in said at least one vegetable oil in a hermetically sealed reactor with a specific combination of temperatures and heating time, it is possible to decarboxylate the prodrugs present in acid form in said at least one inflorescence, and obtain a cannabis oil with a high concentration of THC and CBD and substances which contribute to the therapeutic effect, such as the terpenes, coming from the phytocomplex of the cannabis plant.

The method according to the present invention comprises a step a) of preparing at least one oily dispersion by dispersing at least one unfertilized female cannabis inflorescence in at least one vegetable oil.

The method according to the present invention allows to obtain a cannabis oil starting from unfertilized female cannabis inflorescences having a broad concentration spectrum of active substances, contained therein both as prodrugs in acid form and as therapeutically active substances.

Examples of unfertilized female cannabis inflorescences usable with the method according to the present invention are the products: Bedrocan®, Bedica®, Bedrolite®, Bedrobinol®, Bediol®, FM1®, FM2®, and Pedanios®. Preferably in said step a) the amount of said at least one vegetable oil varies from 2 to 50 millilitres per gram of said at least one cannabis inflorescence. More preferably, the amount of said at least one vegetable oil ranges from 5 to 20 millilitres per gram of said at least one cannabis inflorescence, even more preferably from 8 to 12 millilitres per gram of said at least one cannabis inflorescence.

In the method according to the present invention, any vegetable oil suitable for being used for the production of a cannabis oil for therapeutic use may be used.

Preferably, said at least one vegetable oil is olive oil. Even more preferably, said at least one vegetable oil is O.P olive oil (XII edition of the Official Pharmacopoeia of the Italian Republic).

Preferably, in said step a) said oily dispersion is homogenized, so as to chop said at least one cannabis inflorescence.

The Applicant has in fact found that by subjecting said oily dispersion to a homogenization which chops said at least one cannabis inflorescence, it is possible to increase the quantity of THC and other active substances in the cannabis oil obtained by the method according to the invention.

Said homogenization can be carried out by means of any equipment suitable for the purpose, such as, for example, a turboemulsifier or a homogenizer device by means of ultrasonic sonication. Examples of turboemulsifiers suitable for the purpose are: KT Miccra Homogenizer or RT Miccra D9 Digitron Homogenizer or IKA T25 digital con S25N-25G head. An example of a sonication homogenizer adapted to be used in the method according to the present invention is the sonicator, BAOSHISHAN FS- 600N Ultrasonic Homogenizer 600W Lab Sonicator Processor.

Preferably, said homogenization occurs at a temperature ranging from 0°C to 25°C, even more preferably at a temperature ranging from 0°C to 10°C.

In a preferred embodiment, said homogenization is carried out by placing said dispersion in a container cooled in an ice bath of water at 0°C.

The duration of the homogenization step is preferably between 2 minutes and 15 minutes. The Applicant has also noted that the efficacy of the extraction of the active substances in oil, both as prodrugs in acid form and as therapeutically active substances, contained in the unfertilized female inflorescences can significantly influence the qualitative and quantitative composition of the cannabis oil, in particular by limiting the content of THC and CBD and other active substances deriving from the phytocomplex of the cannabis plant.

Therefore, in a preferred embodiment of the method according to the invention, in said step a) said at least one oily dispersion is stirred in at least one extractor provided with a stirring system, at a temperature ranging from 60°C to 120°C and at a pressure ranging from 2 bar to 40 bar, for a time ranging from 30 minutes to 150 minutes.

Preferably, in said step a) at least one oily dispersion is stirred in at least one extractor provided with a stirring system, at a temperature ranging from 80°C to 120°C, more preferably from 90°C to 110°C, even more preferably from 95°C to 105°C.

Preferably, in said step a) at least one oily dispersion is stirred in at least one extractor provided with a stirring system, for a time ranging from 45 minutes to 130 minutes, even more preferably from 60 minutes to 120 minutes.

In a particularly preferred embodiment, in said step a) said at least one oily dispersion is stirred in at least one extractor provided with a stirring system, at a temperature ranging from 95°C to 105°C and for a time ranging from 100 minutes to 120 minutes.

It has indeed been surprisingly discovered that, by subjecting the at least one oily dispersion to said specific combination of temperature and stirring time, it is possible to increase the content of THC and CBD and of other active substances deriving from the phytocomplex of the cannabis plant, in the cannabis oil.

Preferably, in the method according to the present invention said extractor includes: a hollow body delimiting a seat adapted to contain said at least one oily dispersion, wherein said hollow body has at least one side wall delimiting an opening and a bottom wall; a plug housed on said opening and adapted to hermetically close said seat; - at least one stirring system adapted to stir said at least one oily dispersion inside said seat; a heating system adapted to heat said at least one oily dispersion inside said seat; and at least one valve means adapted to allow the feeding of at least one inert gas inside said seat or apply a vacuum inside said seat by means of a vacuum pump.

Preferably, said extractor includes at least one control unit connected to the stirring system and to the heating system, adapted to send or receive signals therefrom.

Preferably, said stirring system includes a stirbar housed in said seat and a magnetic stirrer or a mechanical stirrer, such as for example a sealed shaft inserted on one of the extractor walls.

In a particularly preferred embodiment, said stirring system consists of a Teflon- coated stirbar placed inside the extractor and a motor and magnetic dipole system placed outside the extractor, in which the extractor has a part transparent to the magnetic field consisting of a material with low magnetic permeance.

Preferably, said heating system includes at least one temperature sensor and at least one heating device selected from the group consisting of: magnetic induction heater, heating plate, water bath device, steam heater, microwave heater (Magnetron). Preferably, the temperature sensor is housed in said seat. Advantageously, the temperature sensor is in contact with said oily dispersion. Alternatively, the temperature sensor can advantageously be an infrared pyrometer, which does not require contact with said oily dispersion. Preferably, said extractor includes a thermal insulation system adapted to thermally insulate said hollow body, comprising at least one covering layer of said hollow body, made of thermal insulating material.

In an embodiment of the present invention, it is also possible to carry out step b) of the method according to the present invention in said extractor.

Preferably, when step b) is carried out using said extractor, an improvement in performance can be obtained by eliminating atmospheric oxygen. This has the advantage of preventing or minimizing the oxidative phenomena of the oily dispersion. The elimination of oxygen is preferably obtained by creating a vacuum in the hermetically sealed extractor, or by injecting an inert gas therein. As inert gas it is possible to use indifferently and equivalently Nitrogen (N2) or a noble gas such as He, Ne, Ar or the like. Preferably, said vacuum is carried out by means of a vacuum pump connected to said at least one valve means of the extractor. In a preferred embodiment, said extractor comprises a condenser, adapted to make the extractor itself usable also as a distiller and extractor of terpenes, terpenoids and flavonoids from raw materials of plant origin. Advantageously, said condenser is applied on said at least one valve means.

Preferably, said condenser is of the Graham condenser type for distillation with counter-current or another type of refrigerant.

When the extractor is used for the extraction of terpenes from raw materials of plant origin, said extractor is used at a temperature between +50°C and +130°C.

In a further preferred embodiment, a filtering and collection accessory is added to the extractor. Said filtering and collection accessory is adapted to separate the oily liquid part of the oily dispersion contained in the extractor.

Preferably, said filtering and collection accessory is applied to the extractor on the opening of the hollow body, after removing the plug, at the end of the extraction step. Preferably, said accessory consists of: a) a flange suitable for being hermetically coupled with said opening of the hollow body; b) a filter, c) at least one fitting preferably provided with a valve means with grafting suitable for connecting a suction pump which extracts the air and creates a vacuum in the collection volume.

In order to use the extractor with said filtering and collection accessory, the extractor is overturned so as to have the filtering and collection accessory below the hollow body, in a position such as to bring the oily dispersion into contact with the filter b) of the accessory.

Preferably, the filter b) is selected in the group consisting of: metal grid filter, paper filter, nylon sock filter, or a combination thereof.

Preferably, said accessory further comprises an oil seal near the at least one fitting c) and adapted to protect said fitting c) from the aspiration of the oily liquid part of the oily dispersion contained in the extractor.

In a preferred embodiment, said at least one fitting c) can further be advantageously used also for the entry of air into the accessory; bubbling in counter-current through the filter b), the air regenerates it cyclically. In a further preferred embodiment, the accessory comprises at least two fittings preferably provided with a valve means, at least one of which is suitable for connecting a suction pump which extracts the air and creates a vacuum in the collection volume, and at least one of which is adapted to introduce air into the accessory; bubbling in counter-current through the filter b), the air regenerates it cyclically.

The method according to the present invention comprises a step b) of heating said oily dispersion to a temperature ranging from 90°C to 150°C and for a time ranging from 20 to 140 minutes, in at least one hermetically closed reactor.

In a preferred embodiment of the method according to the invention, said step b) of the method according to the invention is carried out by irradiating said at least one oily dispersion with a microwave electromagnetic radiation and said reactor of said step b) of the method according to the invention includes at least one wall comprising a part of a material transparent to microwave electromagnetic radiation. Said material transparent to microwave electromagnetic radiation is for example polytetrafluoroethylene, normally known through its commercial names Teflon, Fluon, Algoflon, Hostaflon, Inoflon.

Said material transparent to the microwave electromagnetic radiation is also advantageously selected from the materials suitable for contact with food.

In a further preferred embodiment of the method according to the invention, said step b) of the method according to the invention is carried out by heating said at least one oily dispersion by means of oscillating magnetic induction and said reactor of said step b) of the method according to the invention includes at least one wall comprising a part of an electrically resistive material and/or provided with a magnetic hysteresis loop.

Said electrically resistive material and/or provided with a magnetic hysteresis loop is for example AISI 316 stainless steel, which is also advantageously suitable for contact with food.

In a particularly preferred embodiment of the present invention, the reactor of said step b) of the method according to the invention is an apparatus comprising:

- a hollow body delimiting a seat adapted to contain said at least one oily dispersion, wherein said hollow body has at least one side wall delimiting an opening and a bottom wall which has at least one discharge channel;

- a piston slidably housed in said seat and adapted to hermetically slide against said at least one side wall and to hermetically close said seat;

- at least one fixing element housed at said opening and adapted to lock said piston in abutment and to prevent it from exiting from said seat;

- a filtering element housed in said seat at said bottom wall and adapted to separate said at least one oily dispersion into a solid component and a liquid component; and

- at least one discharge valve means fixed to said bottom wall at said discharge channel. It has in fact been found that the use of such an apparatus is particularly advantageous for obtaining a cannabis oil with a high concentration of THC and CBD and other active substances deriving from the phytocomplex of the cannabis plant.

In fact, thanks to the possibility of using this apparatus, the Applicant has surprisingly discovered that it is possible to obtain the decarboxylation of the THCA and CBDA prodrugs of cannabis to create the therapeutically active forms THC and CBD, to obtain a high content of other active substances deriving from the phytocomplex of the cannabis plant, as well as to rapidly separate at least one liquid component from at least one solid component of the oily dispersion obtained from step d), so as to obtain the cannabis oil.

In a preferred embodiment of the present invention, it is also possible to carry out step a) of the method according to the present invention in said apparatus.

Preferably, the filtering element of the reactor of said step b) includes at least one layer of a mesh filter. Preferably, said at least one layer of a mesh filter is made of a material compatible with food use. Examples of materials suitable to be used to make said at least one layer of a mesh filter are for example AISI-316 stainless steel, OT58 brass, or polytetrafluoroethylene.

Preferably, the piston of the reactor of said step b) includes at least one valve means adapted to allow the feeding of at least one inert gas inside said seat.

Said piston may also optionally also include a safety valve means and an airhole.

Said piston is also advantageously made of a material compatible with food use, such as for example those mentioned above.

Preferably, said at least one valve means for discharging the reactor of said step b) is selected from the group consisting of: ball cock, solenoid valve.

Said at least one valve means is also advantageously made of a material compatible with food use, such as for example those mentioned above. Preferably, the reactor of said step b) includes an actuator adapted to make the piston slide in said seat. Preferably, said actuator is also connected to a locking system capable of locking the sliding of the piston in said seat.

Preferably, the reactor of said step b) includes at least one heating device adapted to heat the oily dispersion inside said reactor.

In one embodiment, said heating device is a device adapted to generate a microwave electromagnetic radiation.

In a further embodiment, said heating device is a device adapted to generate an oscillating magnetic induction to produce heat by exploiting Foucault eddy currents and/or a magnetic hysteresis loop.

Preferably, the reactor of said step b) includes at least one temperature sensor.

Preferably, the reactor of said step b) includes at least one control unit connected to the at least one temperature sensor and to the at least one heating device and adapted to send or receive signals therefrom. Said control unit advantageously allows to regulate the temperature of the oily dispersion, for example during step b) and during step c) of the method according to the present invention.

Preferably, the reactor of said step b) includes at least one partial CO2 pressure sensor. By means of said sensor it is in fact possible to monitor the partial CO2 pressure value and, knowing also the absolute total pressure and the volume of the vapour phase, calculate the CO2 concentration, the total amount of CO2 by weight and therefore, dividing by the molar mass of CO2, the number of moles. Knowing that the amount in moles of CO2 is equal to the amount of decarboxylated CBDA and THCA, it is thus possible to monitor the decarboxylation reaction in real time with good approximation.

When in the method according to the present invention step a) is also carried out in said reactor, said control unit advantageously allows to regulate the temperature of the oily dispersion also in said step a). Preferably, in said step b) said oily dispersion is heated to a temperature ranging from 100°C to 130°C, even more preferably from 110°C to 120°C.

Preferably, in said step b) said oily dispersion is heated for a time ranging from 50 minutes to 110 minutes, even more preferably from 70 minutes to 90 minutes.

The method according to the present invention comprises a step c) of bringing, in the at least one hermetically closed reactor, the at least one oily dispersion obtained from step b) to a temperature ranging from 0°C to 35°C.

Preferably, in said step c) said at least one oily dispersion is brought to a temperature ranging from 0°C to 15°C.

Said step c) can be carried out in any way known to those skilled in the art, for example by spontaneous cooling, radiation, conduction and natural convection. Preferably, in order to make the process faster, said step c) can be advantageously carried out by forced convection through a fan or ice bath or Peltier cells.

The method according to the present invention comprises a step d) of separating at least one liquid component from at least one solid component of the oily dispersion obtained from step c), so as to obtain the cannabis oil.

Preferably, said step d) is carried out by filtering the oily dispersion obtained from step c).

In a further aspect, the present invention relates to an apparatus comprising:

- a hollow body delimiting a seat adapted to contain a dispersion of at least one solid component in at least one liquid component, wherein said hollow body has at least one side wall delimiting an opening and a bottom wall which has at least one discharge channel;

- a piston slidably housed in said seat and adapted to hermetically slide against said at least one side wall;

- at least one fixing element housed at said opening and adapted to lock said piston in abutment and to prevent it from exiting from said seat; - a filtering element housed in said seat at the bottom wall and adapted to separate said dispersion into at least one solid component and at least one liquid component;

- at least one discharge valve means fixed to said bottom wall at said discharge channel; and

- at least one heating device adapted to heat a dispersion of at least one solid component in at least one liquid component contained in said seat, selected from a device adapted to generate a microwave electromagnetic radiation and a device adapted to generate an oscillating magnetic induction to produce heat exploiting Foucault eddy currents and/or a magnetic hysteresis loop; in which said apparatus includes at least one wall comprising a part of a material selected from the group consisting of: material transparent to microwave electromagnetic radiation, and electrically resistive material and/or provided with a magnetic hysteresis loop.

In fact, said apparatus is particularly adapted to be used for the preparation of cannabis oil with the method according to the first aspect of the present invention. The advantages of said apparatus have already been described with reference to the first aspect of the invention and are not repeated here. In a preferred embodiment, the apparatus according to the present invention includes at least one wall comprising a part of any material transparent to microwave electromagnetic radiation, such as for example polytetrafluoroethylene, normally known through its commercial names Teflon, Fluon, Algoflon, Flostaflon, Inoflon.

Said material transparent to the microwave electromagnetic radiation is also advantageously selected from the materials suitable for contact with food.

In a further preferred embodiment of the method according to the invention, the apparatus according to the present invention includes at least one wall comprising a part of any electrically resistive material and/or provided with a magnetic hysteresis loop. Said electrically resistive material is for example AISI 316 stainless steel, which is also advantageously suitable for contact with food.

Preferably, the filtering element of the apparatus according to the present invention includes at least one layer of a mesh filter. Preferably, said at least one layer of a mesh filter is made of a material compatible with food use. Examples of materials suitable to be used to make said at least one layer of a mesh filter are for example AISI-316 stainless steel, OT58 brass, or polytetrafluoroethylene.

Preferably, the piston of the apparatus according to the present invention includes at least one valve means adapted to allow the feeding of at least one inert gas inside said seat.

Said piston may also optionally also include a safety valve means and an airhole.

Said piston is also advantageously made of a material compatible with food use, such as for example those mentioned above.

Preferably, said at least one valve means of the apparatus according to the present invention is selected from the group consisting of: ball cock, solenoid valve.

Said at least one valve means is also advantageously made of a material compatible with food use, such as for example those mentioned above.

Preferably, the apparatus according to the present invention includes an actuator adapted to make the piston slide in said seat. Preferably, said actuator is also connected to a locking system capable of locking the sliding of the piston in said seat.

The apparatus according to the present invention includes at least one heating device adapted to heat a dispersion of at least one solid component in at least one liquid component contained in said seat of the same apparatus.

In one embodiment, said heating device is a device adapted to generate a microwave electromagnetic radiation. In a further embodiment, said heating device is a device adapted to generate an oscillating magnetic induction to produce heat by exploiting Foucault eddy currents and/or a magnetic hysteresis loop.

Preferably, the apparatus according to the present invention includes at least one temperature sensor.

Preferably, the temperature sensor is housed in said seat of said apparatus. Advantageously, the temperature sensor is thermally in contact with said dispersion contained in said seat. Alternatively, the temperature sensor can advantageously be an infrared pyrometer, which does not require contact with said oily dispersion. Preferably, said apparatus includes at least one partial CO2 pressure sensor. By means of said sensor it is in fact possible to monitor the partial CO2 pressure value and, knowing also the absolute total pressure and the volume of the vapour phase, calculate the CO2 concentration, the total amount of CO2 by weight and therefore, dividing by the molar mass of CO2, the number of moles. Knowing that the amount in moles of CO2 is equal to the amount of decarboxylated CBDA and THCA, it is thus possible to monitor the decarboxylation reaction in real time with good approximation.

Preferably, the apparatus according to the present invention includes at least one control unit connected to the at least one temperature sensor and to the at least one heating device, and adapted to send or receive signals therefrom.

Said control unit advantageously allows to regulate the temperature of the dispersion of at least one solid component in at least one liquid component contained in said seat.

In a further embodiment, the present invention refers to a device for extracting terpenes, terpenoids and flavonoids from raw materials of plant origin, including: a hollow body delimiting a seat adapted to contain said at least one oily dispersion, wherein said hollow body has at least one side wall delimiting an opening and a bottom wall; a plug housed on said opening and adapted to hermetically close said seat; at least one stirring system adapted to stir said at least one oily dispersion inside said seat; a heating system adapted to heat said at least one oily dispersion inside said seat; - at least one valve means; and a condenser, wherein said condenser is applied on said at least one valve means.

Preferably, said condenser is of the Graham condenser type for distillation with counter-current or another type of refrigerant. Specific embodiments of the invention will be further described, by way of example only, with reference to the attached figures.

With reference to Figure 1 , this shows a schematic, longitudinal-section view not to scale of an extractor 100 adapted to be used in the method according to the present invention. The extractor 100 includes a hollow body 101, a plug 107, a stirring system 108, a valve means 111, and a control unit 112.

The extractor 100 further includes a heating system 125 which includes a temperature sensor 115, a heating device 130, and a thermal insulation system comprising five layers 118, 119, 120, 121, 122 covering said hollow body 101 made of thermal insulating material. The heating device 130 shown in Figure 1 is a solenoid for magnetic induction, the two blocks 109 and 110 of which are shown in the schematic sectional view.

The hollow body 101 delimits the seat 102 and has two side walls 103, 104 and a bottom wall 106. The two side walls 103, 104 delimit an opening 105, on which the plug 107 is housed, which hermetically closes the seat 102. The valve means 111 is housed on the plug 107, which is adapted to allow the supply of at least one inert gas inside the seat 102 and a safety valve 123. Through the valve means 111 it is possible to supply an inert gas inside the seat 102 hermetically closed by the plug 107, so as to increase the pressure inside the seat 102 itself.

In an embodiment of the extractor 100, not specifically shown in Figure 1 , the extractor 100 comprises a condenser of the Graham condenser type for distillation with counter-current or other refrigerant. In said embodiment, the extractor is adapted to be used as a distiller and extractor of terpenes, terpenoids and flavonoids from raw materials of plant origin of various kinds. Said condenser is advantageously applied on the valve means 111 placed on the plug 107 of the extractor.

The stirring system 108 includes a stirbar 113 housed in the seat 102 and a magnetic stirrer 114, placed at the bottom wall 106 and adapted to make the stirbar 113 rotate on itself around the axis represented by the dotted line 126.

The control unit 112 is connected to the stirring system 108 via the connection 129 and to the heating system 125 and is adapted to send or receive signals therefrom. In Figure 1 , the control unit 112 is connected to the heating devices 109 and 110 of the heating system 125 through the connections 127 and 128.

The control unit 112 is adapted to cause temperature variations of the oily dispersion 124 on the basis of signals sent from the temperature sensor 115 by sending signals to the heating system 125. The heating system 125 regulates the temperature of the oily dispersion 124 through the heating devices 109 and 110.

The seat 102 is adapted to contain the oily dispersion 124 of an unfertilized female cannabis inflorescence in O.P. olive oil (XII edition of the Official Pharmacopoeia of the Italian Republic).

Before switching on the extractor 100, the oily dispersion 124 is fed into the seat 102.

The stirbar 113 is then inserted and the plug 107 is housed on the opening 105, so as to hermetically close said seat 102. The stirring system 108 is then started and the magnetic stirrer 114 rotates the stirbar 113. Rotating on itself around the axis represented by the dashed line 126, the stirbar 113 keeps the oily dispersion 124 under stirring.

An inert gas can be fed into the seat 102 through the valve means 111, so as to increase the pressure inside the seat 102 itself or, alternatively, a vacuum can be applied inside the seat 102 itself by means of a vacuum pump.

When the extractor is used to distil and extract terpenes, terpenoids and flavonoids from raw materials of plant origin of various kinds, a condenser of the Graham condenser type is then applied to the valve means 111 for distillation with counter- current or other type refrigerant.

During the operation of the extractor 100, the heating system 125 is also started which, by means of the heating device 130, heats the oily dispersion 124 contained in the seat 102, causing the temperature thereof to increase, monitored by the temperature sensor 115. The control unit 112 is adapted to determine temperature variations of the oily dispersion 124 on the basis of signals sent from the temperature sensor 115 by sending signals to the heating system 125, which regulates the temperature of the oily dispersion 124 by varying the power of the heating device 130.

Once the use of the heating system 125 is finished, it is turned off and the temperature of the oily dispersion 124 is allowed to gradually drop. Subsequently, the oily dispersion 124 is recovered by the extractor 100 through the opening 105.

Step b) of the process can be carried out in two equal manners, i.e. , in the same extractor shown in Figure 1 provided with an induction heating system or in the container illustrated in Figure 2 provided with a microwave heating system. Figure 5 shows a schematic longitudinal-section view not to scale of an embodiment of an extractor 300 adapted to be used in step a) of the method according to the present invention, which also has a filtering and collection accessory 302.

The extractor 300 is composed of a hollow body 301 , similar to the extractor 100 of Figure 1 , and is shown in Figure 5 upside down and without showing all its parts, for the sake of simplicity. The filtering and collection accessory 302 is applied at the opening of the hollow body 301, which includes: a flange 303 suitable to be hermetically coupled with said opening of the hollow body 301 ; a sealing gasket 304, a metal mesh filter with metal grilles 305, and a fitting 306 equipped with a valve means suitable for connecting a suction pump which extracts the air and creates a vacuum in the collection volume (not shown in Figure 5).

For the use thereof, the filtering and collection accessory 302 is applied to the extractor 300 containing the oily dispersion 307, at the opening of the hollow body 301 , after removing the appropriate plug, at the end of the extraction step. The extractor 300 is then turned upside down, so as to have the filtering and collection accessory 302 below the hollow body 301 , in a position such as to bring the oily dispersion 303 into contact with the filter 305. The fitting 306 is then connected to a vacuum pump which is switched on and generates a pressure gradient such as to allow the separation of the oily liquid phase 308 from said oily dispersion 303. Figure 2 shows a schematic longitudinal-section view not to scale of a hermetically sealed reactor 200 according to the present invention.

The reactor 200 includes a hollow body 201 , a piston 208, a fixing element 225, a filtering element 211, a discharge valve means 212, a linear mechanical actuator 215 adapted to move the piston, a device 217 adapted to generate a microwave electromagnetic radiation, a temperature sensor 218, and a control unit 219.

The hollow body 201 defines a seat 202 and has two side walls 203, 204 and a bottom wall 206. The two side walls 203, 204 delimit an opening 205 at which the fixing element 225 is positioned.

In the apparatus of Figure 2, the fixing element 225 is a ring nut, the two parts 209 and 210 of which are shown in the schematic sectional view.

In the apparatus of Figure 2, the hollow body is cylindrical and entirely made of Teflon.

The bottom wall 206 has a discharge channel 207. The discharge valve means 212 is fixed to said bottom wall 206 at the discharge channel 207. The valve means 213 is housed on the piston 208, which is adapted to allow the supply of at least one inert gas inside the seat 202 and a safety valve 214. The piston 208 is slidably housed in the seat 202 and is adapted to hermetically slide against the side walls 203, 204 and hermetically close the seat 202.

The actuator 215 is adapted to make the piston 208 slide in the seat 202 in the direction indicated by the arrow represented by the reference number 220. The locking system 216 is connected to the actuator, and is adapted to make the piston 208 integral with the actuator 215.

The position of the piston 208 in the seat 202 determines the useful volume of the seat 202 itself.

Through the valve means 213 it is possible to feed an inert gas inside the seat 202 hermetically closed by the piston 208, so as to increase the pressure inside the seat 202 itself.

The control unit 219 is connected to the device 217 through the connection 222 and to the temperature sensor 218 through the connection 223 and is adapted to send or receive signals therefrom.

The seat 202 is adapted to contain an oily dispersion 224 of an unfertilized female cannabis inflorescence in O.P. olive oil (XII edition of the Official Pharmacopoeia of the Italian Republic).

Before switching on the reactor 200, the filtering element 211 is placed in the seat 202 at the bottom wall 206. Before feeding the oily dispersion 224 to the reactor 200, the discharge valve means 212 is brought into the closed position, so as not to allow the contents of the seat 202 to exit through the discharge 207.

Once the oily dispersion 224 has been fed to the reactor 200 and is contained in the seat 202, the piston 208 is housed in said seat 202 and the ring nut 225 is fixed, so as to lock the piston 208 in abutment and prevent it from exiting from said seat 202. The piston 208 hermetically closes said seat 202.

An inert gas can be fed into the seat 202 through the valve means 213, so as to increase the pressure inside the seat 202 itself. During the operation of the reactor 200, the device 217 is switched on and the microwave electromagnetic radiation thus generated irradiates the oily dispersion 224 contained in the seat 202, causing the temperature thereof to increase, monitored by the temperature sensor 218. The control unit 219 is adapted to determine temperature variations of the oily dispersion 224 on the basis of signals sent by the temperature sensor 218 by sending signals to the device 217, which regulates the temperature of the oily dispersion 224 by varying the average value of the power of the microwave electromagnetic radiation over time. In this preferred embodiment of the invention, the control unit 219 consists of a microprocessor suitably programmed with a specific firmware capable of communicating data through a Wi-Fi transmission channel, a WLAN network, software on a PC or tablet or smartphone, provided with a graphic interface.

Once the irradiation of the oily dispersion 224 has ended, the device 217 is switched off and the temperature of the oily dispersion 224 is allowed to gradually drop in a spontaneous or forced manner. Subsequently, the discharge valve means 212 is brought into the open position and the piston 208 is made to slide along the seat 202, moving the actuator 215 towards the bottom wall 206 in the direction indicated by the arrow represented with the reference number 220, thus making the oily dispersion 224 pass through the filtering element 211. The oily dispersion 224 is thus separated into a liquid component which is conveyed through the discharge 207 and collected at a point downstream of the discharge valve means 212, indicated by the arrow represented with the reference number 221, and a solid component which remains inside the seat 202 at the filtering element 211. The separation between the liquid component and the solid component, i.e. , spent inflorescence, may also occur equivalently by means of a filtering system with a standard chemical laboratory vacuum pump.

EXPERIMENTAL PART

The invention is now described by means of some examples to be considered for non-limiting illustrating purposes thereof. EXAMPLE 1

5 grams of Bedrocan® unfertilized female cannabis inflorescences were mixed with 40 millilitres of O.P. olive oil and placed in a Pyrex glass flask.

The oily dispersion obtained was homogenized by using a turboemulsifier MOD. KT MICCRA for 180 seconds inside the flask, so as to chop the cannabis inflorescences, while the container is wrapped with an ice-gel pad previously cooled in the freezer to -20°C, of the Thermogel Artsana (PIC) type.

A Teflon-coated stirbar was then placed on the bottom of the flask and the flask was closed by positioning the appropriate silicone plug in place, hermetically sealing the system.

The flask was placed on a magnetic thermo-stirrer at a temperature of 100°C for 120 minutes.

Subsequently, the flask was cooled in an ice bath and the contents were poured into a Teflon container shaped as in Figure 2, which was inserted into a Whirlpool ChefPlus Mod.MCP345 800W microwave oven suitably modified with the addition of a control module which detects the oil temperature by means of a thermocouple and controls the emission of microwaves.

The Teflon container is equipped with a discharge valve with a ball cock, kept in the closed position so as to prevent the oily dispersion from exiting. After feeding the oily dispersion into the container, the special piston was housed in the seat containing the oily dispersion itself, then fixing the ring nut so as to stop the piston from exiting and thus hermetically sealing said seat.

Subsequently, the apparatus was irradiated with a microwave electromagnetic radiation, so as to heat the oily dispersion to a temperature of 116°C for a time of 80 minutes.

The oily dispersion was then allowed to cool down to 25°C.

Subsequently, the ball valve was put in the open position, the piston was pushed towards the bottom wall, and the oily dispersion exiting was collected to be subsequently filtered through a filtering system with a vacuum pump, so as to obtain the cannabis oil.

The resulting cannabis oil was analysed by means of GC-MS using an Orbitrap qExtractive mass spectrometer to determine the content of THC, CBD, THCA, CBDA and other non-cannabinoid active substances. The results are shown in Table 1.

The present method was replicated 5 more times and the results of said 5 replications are also shown in Table 1 , identified respectively with the words “Example 1 - replication 1”, “Example 1 - replication 2”, “Example 1 - replication 3“, “Example 1 - replication 4”, and “Example 1 - replication 5”.

TABLE 1

EXAMPLE 2

5 grams of Bedrocan® unfertilized female cannabis inflorescences were mixed with 40 millilitres of O.P. olive oil and placed in a stainless steel container shaped like the apparatus of Figure 1.

The oily dispersion was then homogenized by using a turboemulsifier MOD. KT MICCRA for 180 seconds inside the container, so as to chop the cannabis inflorescences, while the container was wrapped with an ice-gel pad previously cooled in the freezer to -20°C, of the Thermogel Artsana (PIC) type.

A Teflon-coated stirbar was placed on the bottom of the container.

The container was hermetically closed. The vacuum was then created inside the container by means of a commercial vacuum pump capable of reaching a vacuum of 50 Pa.

The container was inserted into a magnetic induction heating system with an integrated magnetic stirrer which keeps the stirbar rotating and provided with temperature control.

The temperature control system was programmed to impose the following temperature/time profile on the oily dispersion: 100°C for 120 minutes and then 116°C for 80 minutes.

At the end of this process, the container was cooled by an ice bath and the contents were poured into the filtering system with a vacuum pump.

The cannabis oil thus obtained was analysed using the same method used in Example 1 to determine the content of THC, CBD, THCA, CBDA and other non- cannabinoid active substances.

The results obtained are shown in the following TABLE 2. The present method was replicated 5 more times and the results of said 5 replications are also shown in Table 2, identified respectively with the words “Example 2 - replication 1”, “Example 2 - replication 2”, “Example 2 - replication 3“, “Example 2 - replication 4”, and “Example 2 - replication 5”.

TABLE 2

The efficiency of the method was confirmed by the high amounts of THC obtained through the indicated method, and by the completeness of the phytocomplex highlighted by the gas-chromatographic fingerprint performed with replicas 2 and 3, the chromatograms of which are shown respectively in Figures 3 and 4. In the cannabis oil samples obtained with replicas 2 and 3 a significant presence of terpenes was detected (Figures 3 and 4, retention time interval from 0 to 60 minutes), among which in particular evidence we find: pinene, carene, caryophyllene, and myrcene. Furthermore, from the comparison of the two figures 3 and 4 it is possible to note that the replicas 2 and 3 have essentially the same components in the same proportions.

In Figures 3 and 4, in the retention time interval from 80 to 115 minutes, compounds from the oil used for the extraction were detected, i.e. , oleic acid and squalene, in addition to the cannabinoids: CBD (retention time: 97 minutes), TFIC (retention time: 102 minutes), cannabinol (retention time: 104 minutes).

The two chromatograms obtained from the analysis of replicas 2 and 3 showed that the phytocomplex present in the oleolyte was preserved and was comparable with what was detected by analyses carried out on unfertilized female cannabis inflorescences available on the market under the commercial names Bedrocan and Pedanios, performed by the University of Milan by means of solid phase micro extraction coupled with gas chromatography and mass spectrometry (FIS-SPME- GC/MS), and presented at the Canapaforum 2018 Research Innovation Development event (Saturday 27 and Sunday 28 October 2018). This confirmed the substantial maintenance of the cannabis phytocomplex through the method according to the present invention.

Finally, from the results obtained in Examples 1 and 2, it is possible to appreciate how the method according to the present invention allows to obtain greater TFIC concentrations than those obtainable using the known extraction methods: Romano Flazekamp method, Cannazza method, and SIFAP method.

The following Table 3 shows the comparison between the yields of different preparations carried out with the different methods using the same Bedrocan® unfertilized female cannabis inflorescences, used in Examples 1 and 2 according to the present invention. TABLE 3

* data presented by the State University of Milan, Pharmacy Dept., Laboratory Managers Researchers Gabriella Roda, Veniero Gambaro, Eleonora Casagni at the “First Italian Conference on Cannabis as a possible drug” event of 15 May 2018.

As it can be noted, in Examples 1 and 2 according to the present invention, the THC concentration values in the cannabis oil are obtained at around 2%, values well above the values obtained with the SIFAP, Cannazza and Romano-Hazekamp methods. As evidenced in the present experimentation, the method according to the present invention allows to obtain higher THC concentrations in the cannabis oil, while simultaneously maintaining the phytocomplex of the plant.