EXTRACTION AND STABILIZATION OF PHYTO CANABINOIDS AND TERPENS ON THE BASIS OF LIPIDE ASSISTED BY ENZ

专利摘要:
the present application provides a safe, efficient, environmentally safe and convenient method for extraction and stabilization based on lipids, aided by enzymes, phyto-cannabinoids and terpenes / terpenoids from plant materials. the lipid-based extract has a high content of cannabinoids and increases the stability of phyto-cannabinoids and terpenes / terpenoids, and can be valued in multiple applications in the health, cosmetics, food and agriculture sectors.
公开号:BR112019014463A2
申请号:R112019014463-3
申请日:2018-01-15
公开日:2020-02-11
发明作者:Venturini del Greco Giovanni
申请人:Herbolea Biotech S.R.L.；
IPC主号:

专利说明:

EXTRACTION AND STABILIZATION OF PHYTO CANABINOIDS AND TERPENS ON THE BASIS OF LIPID ASSISTED BY ENZYME AND PRODUCTS OBTAINED
BACKGROUND [0001] Cannabis refers to a genus of annual herbaceous plants in the family Cannabaceae. Human beings have cultivated Cannabis throughout recorded history as a source of industrial fiber, seed oil, food and medicine and for religious, spiritual and recreational purposes.
[0002] Although the main psychoactive component of Cannabis is tetrahydrocannabinol (Δ9-ΤΗΟ, the plant is known to contain more than 400 compounds, including at least 60 cannabinoids. In addition to THC, another cannabinoid produced in high concentrations by some plants is cannabidiol (CBD). CBD is considered to have a broad scope of potential medical applications - due to clinical reports showing the absence of side effects, particularly the lack of psychoactivity (as is typically associated with A9-THC) and not interference with various functions of psychomotor and psychological learning In addition to cannabinoids, hemp or cannabis contain several terpenes, such as terpineol, limonene, myrcene, terpinolene, humulene and sesquiterpenes. When terpenes are chemically modified, this is as by oxidation or rearrangement of the carbon skeleton, the resulting compounds are generally referred to as terpenoids. es are terpenoids, but not all terpenoids are cannabinoids. Terpenes and terpenoids, including
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2/52 cannabinoids, are generally supportive substances and, therefore, soluble in lipids. Some authors use the term terpene more widely, to include terpenoids.
[0003] Terpenes exhibit unique therapeutic effects that can contribute significantly to the effects of grouping cannabis-based medicinal extracts. It was hypothesized that some terpenes could mitigate the undesirable effects of THC (Russo, 2011). Terpenes, not cannabinoids, are responsible for the aroma of cannabis. Monoterpenes generally predominate (limonene, mircene, pinene), but these free space volatiles (Hood et al., 1973), although only lost at a rate of about 5% before processing (Gershenzon, 1994), suffer decreased yields with drying and storage (Turner et al., 1980; Ross and ElSohly, 1996), resulting in a higher relative proportion of sesquiterpenoids (especially karyophylene), as also occurs frequently in extracts.
[0004] The synergistic interaction of different cannabinoids and terpenes is known as the clustering effect introduced in the science of cannabinoids in 1998 by S. Ben-Shabat, with Raphael Mechoulam, to represent a new route of molecular regulation of endogenous cannabinoids.
[0005] The endocannabinoid system consists of endogenous cannabinoids (endocannabinoids), cannabinoid receptors and enzymes that synthesize and degrade endocannabinoids. Many of the effects of cannabinoids and endocannabinoids are mediated by two G protein-coupled receptors (GPCRs), CB1 and CB2, although receptors
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3/52 additional may be involved. CB1 receptors are present at very high levels in various regions of the brain and in smaller amounts in a more widespread manner. These receptors mediate many of the psychoactive effects of cannabinoids. CB2 receptors have a more restricted distribution, being found in several immune cells and in some neurons. CB1 and CB2 are mainly coupled to inhibitory G proteins and are subject to the same pharmacological influences as other GPCRs. Thus, partial agonism, functional selectivity and inverse agonism play an important role in determining the cellular response to specific cannabinoid receptor ligands.
[0006] Interaction with the endocannabinoid system, exogenous cannabinoids, such as cannabis, are used to reduce nausea and vomiting during chemotherapy, to improve appetite in people with HIV / AIDS and to treat chronic pain and muscle spasms. Cannabis, its constituent cannabinoids and terpenes are used to treat disease or improve symptoms. Cannabinoids are being researched preliminarily for their potential to affect stroke or childhood epilepsy. Cannabis sativa L. is a prolific, but not exclusive, producer of a diverse group of isoprenylated resorcinyl polyketides collectively known as cannabinoids (Hanus et al., 2016), and cannabis cannabinoids are the only exogenous lipid compounds that interact with the endocannabinoid system. In recent years, other plants have been found to produce cannabinoid-like compounds and various natural products from non-cannabinoid plants
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Traditional 4/52 have been reported as cannabinoid receptor ligands. Consequently, a natural plant-derived product capable of interacting directly with cannabinoid receptors or sharing chemical similarity with cannabinoids or both is described as 'phytocannabinoid' (Gertsch et al., 2010).
[0007] Among phyto-cannabinoids other than traditional cannabis cannabinoids, which have been reported to interact with the endocannabinoid system, Nalkylamides (N-alkylamides) of unsaturated fatty acids from the medicinal plant Echinacea, a species of perennial herbaceous plant of the family Asteraceae, demonstrated that endogenous cannabinoids bind more strongly to the CB2 receptor (Raduner et al., 2006 and Woelkart et al., 2005). The interaction of N-alkylamides with the endocannabinoid system proved to modulate the induced immune response. Other constituents of purple Echinacea act as weak CB1 antagonists (Hohmann et al., 2011).
[0008] Salvinorin A, a diterpene in Salvia divinorum, produces CB1-mediated effects on the gastrointestinal tract of rodents. Salvinorin A acts mainly as a kappa-opioid receptor agonist and is inactive as a ligand for CB1 and CB2 (Capasso et al., 2008); can interact with a putative CBl-kappa-opioid receptor heterodimer (Fichna et al., 2012).
[0009] Bitter acid humulone, a terpenoid contained in Humulus lupulus (hops) from the same Canabinaceae family, is considered responsible for the sedative effect. Humulone is subject to degradation in about 12 weeks at room temperature (Darby, 2015).
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Curiously, in 2017, The Isodiol, an company of biotechnology, claimed to have extracted CBD of strains modified hops. [0010 ] Hyperforin is also one terpenoid produced by herb of Are John (Hypericum perforatum).
Cannabinoids, bitter acids and hyperforins are polyketides with terpenoid blocks and are lipophilic due to their terpenoid portion (Osburn and Lanzotti, 2009).
[0011] Pyrethrins are terpenoids produced by plants of the genus Chrysanthemum. Pyrethrins can be found in cannabis plant material and extracts, since pyrethrins are used in the formulation of natural pesticides for protection against cannabis. Interestingly, in 2017, Devitt-Lee and others, reported the possibility that pyrethrins can be endogenously synthesized by cannabis, so they can become an additional component of cannabis extractable phyto-complex.
[0012] Among the plants that produce cannabinoid-like compounds, Helichrysum umbraculigerum, a South African species of the ever-living, is also a major producer of CBG (Bohlmann et al., 1979). Other plants that contain cannabinoid-like compounds are Chinese rhododendron and liver radula Marginata in New Zealand (Toyota et al., 2002).
[0013] Several patents and scientific publications are available, providing different dosages of phytocannabinoids for treating diseases or symptoms. For example, doses in the range of 10-20 mg / kg / day of CBD are expected to treat epilepsy (GB2548873). For
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6/52 person weighing 50 kg, that would mean 500-1,000 mg / day of CBD. Assuming a concentration of 4% CBD in the extract, this would require a daily assumption of 12.5-23 mL of extract. In the case of 0.5% concentration of CBD in the extract, the dosage of the extract would be in the range of 100-200 mL, a significant amount to be consumed daily.
[0014] Various processes to extract phytocannabinoids and / or terpenes / terpenoids have been developed. The following main extraction processes are known:
1. Cold pressing for the production of hemp seed oil. Hemp seed oil is rich in nutrients and is a good addition to any diet, but contains only small amounts of cannabinoids (<2%, in the case of industrial hemp), since it is made only with the seeds of the plant. Hemp seed oil can certainly be added to CBD supplements as a basis for these products. However, cold pressing is not useful for producing a cannabinoid-rich oil, as cannabinoids are mainly contained in stems and buds that cannot be processed directly by a normal press or expeller.
2. The Rick Simpson Method for Cannabis Oil is a popular extraction method for extracting CBD oil, which uses petroleum or naphtha as a solvent. This method, although efficient in extracting the active compounds from the cannabis plant (usually made with plants rich in THC), generally leads to products with a lower concentration of terpenoids and other cannabinoids such as CBD, while
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7/52 effectively produces higher concentrations of THC. The main disadvantage of such a method is that solvent residues can remain and potentially interfere with immune function, as described by Romano and Hazekamp (Cannabis Oil: Chemical evaluation of na upcoming canabisbased-medicine 2013).
3. Ethanol extraction can be used to extract the entire range of cannabinoids from the cannabis plant, and is safer than the Rick Simpson method. On the other hand, ethanol has low selectivity and extracts chlorophyll and undesirable waxes, so that the final product has an unpleasant taste. Chlorophyll can be removed by filtering the extract, but this additional step also removes a significant proportion of cannabinoids, leading to a less potent extract. In addition, the stability of cannabinoids, as well as N-alkylamides in ethanolic extracts, is low (Citti et al., 2015 and Spelman, 2009).
4. Extraction with Sonication / ultrasonic waves: C. Da Porto, (Ultrasound-assisted extraction of volatile compounds from industrial Cannabis sativa L. Inflorescences, 2014) describes the procedures for extracting THC and hemp terpenes using ultrasonic waves. The use of ultrasound increased THC extraction, but after 15 min. of treatment the overall extraction efficiency has not yet been satisfactory.
5. Supercritical CO2 extraction (US 9186386 B2, US 6403126 Bl) can be an efficient method to obtain a highly enriched cannabinoid oil (> 60%). At such a concentration level, the product is not directly
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8/52 consumed, but is diluted with vegetable oils, such as olive oil, to reach 3-5%. The method uses safe solvents, but requires complex equipment and expertise, in addition to requiring high energy, so the product obtained is very expensive, making the potential health benefits obtained thanks to the therapeutic use of cannabinoids not accessible to everyone. In addition, it requires that the initial cannabis material be dried, adding a time-consuming step and having negative effects on important compounds, such as volatile monoterpenes. In addition, the process itself is subject to significant losses in terms of monoterpene extraction yield, making it difficult to group the extracts. In addition, it has high selectivity for toxic components that may be present in pesticides, so there may be a risk associated with its presence in a concentrated form in the final product. In addition, the CO2-SC extraction product may have a chemotypical impression significantly different from the cannabis flower (Sexton, 2017). Finally, the stability of cannabinoids extracted with CO2 diluted in olive oil is lower than that obtained with their direct extraction in olive oil, as described by Cannazza (Medicinal cannabis: Principal canabinoids concentration and their stability evaluated by a highperformance liquid chromatography coupled with diode array and quadrupole time off light mass spectrometry method, 2016).
6. Microwave extraction. Koturevic et al., (A rapid method for the extraction of cannabionoids from cannabis sativa using microwave heating technique, 2014) described the possibility of using microwaves to assist
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9/52 the extraction of cannabinoids by organic solvents. Few organizations, such as the NBIRC (New Brunswick Innovation Research Chair) in Medical Technologies, Radient Technologies and Scientus Pharma have announced partnerships with cannabis producers to develop microwave-assisted cannabinoid extraction methods. The technical data are still limited, however technical limitations may derive from the step of separating the solvent from the plant material, the recovery of the solvent that remains adsorbed on the plant matrix, the solvent ratio for the plant material and, finally, the possibility of reaching high concentration in extracts, if non-volatile solvents are used (eg vegetable oils).
7. The Romano-Hazekamp method is based on the extraction of cannabinoids from dry and preheated cannabis inflorescences using vegetable oils (ie olive oil) as solvents. The method can be used to extract the entire range of cannabinoids from the cannabis plant and has the advantage of being very safe for consumption. In addition, it is considered the most environmentally sustainable process. (Cannabis Oil: chemical evaluation of an upcoming cannabis-based medicine, Luigi L Romano, Arno Hazekamp, 2013). The disadvantages of this simple and increasingly popular method are that, in order to obtain a satisfactory cannabinoid extraction yield, extraction with vegetable oils must take place at 98 ° C for a prolonged time (1-2 h) and the amount of oil for to be added as a solvent to the plant material is 4 to 10 times the amount of plant material, consequently, the cannabinoid content in the attainable oil is less than 1% and more than
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50% of volatile monoterpenes are lost due to prolonged treatment at high temperature. Finally, the stability of cannabinoids in vegetable oil is very low, with a degradation in just two weeks of more than 15% and more than 20% for storage at 4 ° C and room temperature, respectively, as described by Pacific! ((Evaluation of canabinoids concentration and stability in standardized preparations of cannabis oil by ultra-high performance liquid chromatography tandem mass spectrometry 2017).
8. Steam distillation and hydrodistillation are traditional methods of extracting monoterpenes. Steam distillation involves hanging a basket of herbs above a container of boiling water. The steam passes through the perforated basket and enters the plant material. Only volatile compounds, such as monoterpenes, are soluble in steam. Hydrodistillation is similar to steam distillation, except that the herb is placed directly in boiling water. The methods are not suitable
for substances not volatile, such as cannabinoids or more terpene compounds heavy. [0015] So, it is necessary to develop new and more efficient processes for extraction and stabilization in
phyto-cannabinoids and / or lipophilic terpenes / terpenoids, even from freshly harvested plant material, using green and / or food solvents and excipients, such as vegetable oils and the or other of these lipids.
SUMMARY OF THE INVENTION [0016] This document provides a useful process for lipid-based extraction aided by
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11/52 enzyme of lipophilic cannabinoids and / or terpenes / terpenes from plant material, such as hemp, cannabis, hops, echinacea, sage dinivorum, chrysanthemum, helichrysum and hypericum biomass. Thus, in one aspect, the document provides a process for producing a fat-soluble extract from plant material containing phytocannabinoids and terpenes / terpenoids, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with enzymes to form a mixture to which water and lipids are optionally added;
ç. stirring the mixture at a temperature between 1 and 80; and
d. separating the mixture into a lipid phase, an aqueous phase and a solid phase;
wherein the lipid phase comprises the fat-soluble extract.
[0017] The process according to the present invention can be carried out by proceeding with steps a. and
B. only, preserving the mixture resulting from step b. and proceeding with the addition of lipids and the separation step d. subsequently, with or without stirring the mixture. The separation step d. it can be performed after one or more days, even in a different laboratory or facility.
[0018] In another aspect, this document provides a fat-soluble extract that can be obtained from the process according to the invention, in which the extract has a total phyto-cannabinoid content of at least 2, 3, 4 or
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12/52 weight percent.
[0019] In yet another aspect, this document provides a lipid-soluble extract that can be obtained from the process according to the invention, in which the ratio between the two main cannabinoids in the fat-soluble extract differs by less than 10%, preferably less than 5%, of the ratio between the two main cannabinoids in the plant material.
[0020] In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, in which less than 10%, preferably less than 5%, more preferably less than 2% of cannabinoids are decarboxylated. during the process.
[0021] In yet another aspect, the present document provides that the stability of cannabinoids in the lipid phase is significantly increased by maintaining at least 90% of the cannabinoid content after two weeks at room temperature. In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, wherein the extract has a terpenoid content of at least 75% of the initial content of plant material by weight.
[0022] In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, in which the monoterpene content is at least 30% of the total terpene content.
[0023] In yet another aspect, this document provides that lipids can be vegetable oils
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13/52 and / or glycerin and / or any other green solvents (obtained from renewable sources) and / or food grade.
[0024] In yet another aspect, this document provides a solid extract or a phase of cannabis or hemp plant material that can be obtained from the process according to the present invention, wherein the cannabinoid content of the plant material is reduced by at least 75%, 80% or preferably 90% by weight.
[0025] In yet another aspect, the present document provides the use of the solid extract or phase of cannabis or hemp plant material that can be obtained from the process according to the present invention for the formulation of food or feed.
[0026] In another aspect, the aqueous phase can be used in the production of nutraceuticals, antimicrobials, antibacterials or biopesticides.
[0027] In yet another aspect, the present document provides that the use of fat-soluble extract for the preparation of a cream containing at least 0.5% cannabinoids presenting an increased stability of
cannabinoids from at least 90% of initial content after 10 weeks. [0028] Still in other aspect, the gift document provides the use of extract fat soluble for The preparation in a gum containing at least 0.5% in
cannabinoids exhibiting increased cannabinoid stability of at least 90% of the initial content after 10 weeks.
[0029] In yet another aspect, the present
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14/52 this document provides for the use of fat-soluble extract for the preparation of a gel containing at least 0.5% cannabinoids exhibiting an increased cannabinoid stability of at least 90% of the initial content after 10 weeks.
[0030] In yet another aspect, this document provides a liposome-based material containing at least 0.5% cannabinoids, exhibiting an increased cannabinoid stability of at least 90% of the initial content after 10 weeks.
DETAILED DESCRIPTION OF THE INVENTION [0031] The present invention discloses a safe, ecological and convenient method for extracting phytocannabinoids and lipid-based terpenes / terpenoids.
[0032] The present invention discloses a process for producing a lipid-soluble extract from plant material containing phyto-cannabinoids, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with enzymes to form a mixture to which water and lipids or solvents can optionally be added;
ç. stirring the mixture at a temperature between 1 and 80 ° C; and
d. separating the mixture into a lipid phase, an aqueous phase and a solid phase;
wherein the lipid phase comprises the fat-soluble extract.
[0033] The present invention further describes a process for the production of a fat-soluble extract
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15/52 containing cannabinoids and / or terpenes / terpenoids from plant material, chosen from the group consisting of hemp, cannabis, hops, echinacea, sage dinivorum, chrysanthemum, helichrysum and hypericum biomass, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with enzymes to form a mixture to which water and lipids or solvents can be optionally added;
ç. stirring the mixture at a temperature between 1 and 80 ° C; and
d. separation of the mixture in a lipid phase, a
aqueous phase and a solid phase; on what the lipid phase comprises the extract fat-soluble. [0034] A lipid fraction generated has a content in phyto-cannabinoid (i.e. CBD, CBD-A, THC , THC-A) in
at least 0.1%, preferably 2%, more preferably 3%, even more preferably at least 4%. Phyto-cannabinoids containing plant material, such as hemp or cannabis, can be fresh or dry. The plant material is ground to increase surface contact. Then, water, enzymes and oil are added to the plant material to form a homogeneous mixture or mud; Temperature and pH conditions may vary according to the specific enzyme or enzyme cocktail used to dissolve the plant material. The mixture can be mixed by stirring or other stirring methods for at least 30 min. to let enzymes degrade plant material. Ultrasound / sonication or microwave or
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16/52 steam blast can be used before or after adding enzymes to the mixture to reduce the time needed to achieve the dissolution of plant material and the high lipid extraction yield of cannabinoids. The proportion of water to plant is important to achieve degradation of plant material through enzymatic activity; freshly harvested plant material can also be used directly, avoiding the pre-drying step during which degradation and / or loss of phytocannabinoids and terpenes, especially monoterpenes, may occur; in that case, little or no water can be used. Lipids can be added to the mixture at any time without significantly modifying the enzyme activity; an adequate proportion of lipid material for plant to obtain high phyto-cannabinoid content and high extraction yield (at least 70%, preferably at least 80%, more preferably at least 90%) is in the range of 50 to 200%, preferably 50 to 150%. The obtained mixture is then separated by density separation (ie, centrifugation) or pressing (French press) and / or filtration to recover a lipid fraction highly enriched with cannabinoids and free waxes. In the case of lipid extract obtained from cannabis, the extract can be heated to form cannabinoids in the desired form of decarboxylate acid.
[0035] Surprisingly, it has been found that the use of enzymes dramatically increases the extraction of phytocannabinoids and terpenes / terpenoids based on lipids, including volatile monoterpenes, allowing for a significant reduction in the solvent-plant lipid ratio (ie 10
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17/52 times compared to the traditional RomanoHazekamp method), although they still achieve a high cannabinoid extraction yield (90%), hence the possibility of obtaining a safe and direct wax-free lipid extract with a phyto-cannabinoid content and terpene suitable and compatible with the dosage of therapeutic applications, where the fingerprint of the terpene of plant material is faithfully reproduced. In addition, it has also been discovered that the use of enzymes dramatically increases the stability of phyto-cannabinoids and terpenes / terpenoids in the extract, allowing to reach an appropriate validity and compatible with pharmaceutical applications without the addition of preservatives.
[0036] In addition, the solid fraction generated by the process shows a significantly reduced phyto-cannabinoid content. In the case of hemp seeds, the cannabinoid content was considerably reduced compared to the mechanical bagasse, making the solid fraction rich in proteins compatible with the safety guidelines for applications in food and food products.
[0037] Aspects of the embodiments of the present disclosure are described in more detail below.
DEFINITIONS [0038] Listed below are definitions of various terms used to describe this invention. These definitions apply to terms as they are used throughout this specification and claims, unless otherwise limited in specific cases, individually or as part of a larger group.
[0039] Unless otherwise defined, all technical and scientific terms used herein
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18/52 document generally have the same meaning as that normally understood by one skilled in the art to which this invention belongs. Generally, the nomenclature used in this document and laboratory procedures in cell culture, molecular genetics, organic chemistry and peptide chemistry are well known and commonly used in the art.
[0040] As used in this document, articles a, o and one, one refer to one or more of one (that is, at least one) of the grammatical object of the article. For example, an element means an element or more than one element. In addition, the use of the term including, as well as other forms such as include, include and included, is not limiting.
[0041] As used herein, the term cannabinoids includes, but is not limited to, cannabinol (CBN), cannabinolic acid (CBNA), Δ (9) tetrahydrocannabinol (A (9) -THC), Δ (9) tetrahydrocannabinolic acid (Δ (9) -THCA), Δ (9) - cannabidiol (Δ (9) -CBD), Δ (9) - tetrahydrocannabidiolic acid (Δ (9) CBDA), Δ (8) -tetrahydrocannabinol (A (8) -THC), Δ (8) tetrahydrocannabinolic acid (Δ (8) -THCA), Δ (8) tetrahydrocannabidiol (Δ (8) -CBD), Δ (8) tetrahydrocannabidiolic acid (Δ (8) -CBDA), Δ (9) tetrahydrocannabivarin (Δ (9) -THV), cannabigerol (CBG), cannabigerolic acid (CBGA), cannabichromene (CBC), cannabichromic acid (CBCA), cannabicyclol (CBL), cannabicyclic acid (CBLA), cannabidivarin (CBDV) and tetrahydrocannabivarin (THCV).
[0042] N-alkylamides include, but are not
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19/52 limited to dodeca-2E, 4E, 8Z, 10Ztetraenoic acid isobutylamide and dodeca-2E acid isobutylamide, 4Edienic acid.
[0043] As used herein, the term phyto-cannabinoids includes, but is not limited to, Cannabis cannabinoids and Echinacea N-alkylamides.
[0044] As used herein, the term terpenes includes, but is not limited to, pinene, limonene, terpinene, terpinen-4-ol, carvacrol, carvona, 1,8-cineol, p-cymene, fenchone, β-myrcene , canaflavin A, canaflavin B, nerolidol, phytol and squalene.
[0045] As used herein, the term terpenoids includes, but is not limited to, cannabinoids, limonene oxide, pulegone-1,2-epoxide, salviorin A, hyperforin and pyrethrins.
[0046] As used herein, the term lipids includes, but is not limited to, olive oil, coconut oil, vegetable oil, milk, butter, liposomes, glycerin, polyethylene glycol, ethyl acetate, d-limonene, butylene glycol, propylene glycol, ethylhexyl palmitate.
[0047] As used in this document, the term about will be understood by those versed in the common technique and will vary to some extent in the context in which it is used. As used in this document, when referring to a measurable value such as a quantity, a time duration and the like, the term about is intended to cover variations of + 20% or + 10%, including + 5%, + 1%, and + 0.1% of the specified value, as such variations are appropriate to carry out the revealed methods.
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Process and Compositions:
[0048] In one aspect, this document provides a process for producing a fat-soluble extract containing cannabinoids and terpenoids from cannabis or hemp plants or plant material containing phytocannabinoids and / or terpenes / terpenoids, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with enzymes to form a mixture to which water and lipids or solvents can be optionally added;
ç. stirring the mixture at a temperature between 1 and 80 ° C; and
d. separating the mixture into a lipid phase, an aqueous phase and a solid phase;
wherein the lipid phase comprises the fat-soluble extract.
[0049] In a preferred aspect of the process according to the present invention, said cannabis or hemp is chosen from the group consisting of buds, flowers, leaves, stems, roots and seeds or a mixture thereof. In one embodiment, the plant material includes seeds. In another embodiment, when the plant material includes seeds, no lipid is added. In another embodiment, when the plant material includes seeds, a lipid is added. Plant material including seeds may be rich in lipids, and thus may not need the addition of lipids.
[0050] In one embodiment, the plant material is a mixture comprising buds, flowers, leaves,
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21/52 stems, stems, roots and seeds. In another embodiment, when the plant material is a mixture comprising buds, flowers, stems, leaves, roots and seeds, a lipid is added to obtain an optimal proportion of lipid-to-plant material for the effective extraction of cannabinoids. In another embodiment, when the plant material is a mixture comprising seeds, buds, flowers, stems, roots and leaves, a lipid is not added.
[0051] In one embodiment, the plant material is freshly harvested and contains a high level of moisture; in that case, adding extra water to the plant material is unnecessary.
[0052] In one embodiment, the plant material has a lipid content of at least 1% by weight.
[0053] In another embodiment, the addition of lipids reaches a lipid content of at least 5 percent by weight of the mixture.
[0054] In a preferred aspect, in the process according to the present invention, said plant material containing phyto-cannabinoids is derived from cannabis, echinacea or hemp plants that are pure, hybrid or their genetically modified variants.
[0055] Still in a preferred aspect, in the process according to the present invention, said plant material derives from the genus Cannabis of plants, which includes the species C. sativa, C. indica and C. ruderalis. Said cannabis or hemp is preferably industrial hemp of the species C. sativa.
[0056] Still in a preferred aspect, in the process
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22/52 according to the present invention said plant material derives from the plant genus Canabinaceae, which includes the species Cannabis and Humulus lupulus.
[0057] Still in a preferred aspect, in the process according to the present invention said plant material derives from the plant genus Echinacea, which includes the species E. purpurea, E. angustifolia, E. pallida.
[0058] Still in a preferred aspect, in the process according to the present invention, said plant material derives from the genus Chrysanthemum, which includes the species Tanacetum cinerariifolium and Chrysanthemum coccineum.
[0059] Still in a preferred aspect, in the process according to the present invention, said plant material is derived from Salvia divinorium.
[0060] Still in a preferred aspect, in the process according to the present invention, said plant material is derived from a mixture of different plant materials containing different terpenes / terpenoids.
[0061] In one embodiment, in the process according to the present invention, said plant material contains at least 0.1, 1 or 2% phyto-cannabinoids by weight.
[0062] In one embodiment, in the process according to the present invention, said plant material contains at least 0.5% terpenoids by weight.
[0063] In one embodiment, the plant material is hemp comprising less than 0.2% - 0.6% THCtot.
[0064] In yet another embodiment, the
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23/52 plant material is cannabis comprising more than 0.2% 0.6% THCtot.
[0065] In another embodiment, the plant material is a hybrid or genetically modified variant of hemp.
[0066] In yet another embodiment, the plant material is a hybrid or genetically modified variant of cannabis.
[0067] In another aspect, in the process according to the present invention, said plant material has a moisture content of at least 20%, preferably at least 30%. In addition, the plant material preferably has a total cannabinoid content greater than 0.2%, preferably greater than 1%.
[0068] In one embodiment, the enzyme is one or more enzymes selected independently from the group consisting of cellulase, beta-glucosidase, hemicellulase, xylanase, glucanase, beta-glucanase, pectinase, amylase, alpha-amylase, phospholipase, beta- mannanase, arabinanase, phytase and protease. In one embodiment, the enzyme is cellulose. In another embodiment, the enzyme is betaglycosidase. In another embodiment, the enzyme is hemicellulase. In another embodiment, the enzyme is xylanase. In yet another embodiment, the enzyme is glucanase. In yet another embodiment, the enzyme is pectinase. In yet another embodiment, the enzyme is amylase. In yet another embodiment, the enzyme is phospholipase. In yet another embodiment, the enzyme is arabinanase. In yet another embodiment, the enzyme is phytase. In another embodiment, the enzyme is protease.
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24/52 [0069] In a preferred embodiment, the enzyme is a mixture or cocktail of cellulase, beta-glucanase, pectinase, beta-mannanase, alpha-amylase and protease; wherein the amount of enzyme is 3% by weight of the plant material; and the pH of the mixture is adjusted to pH 5.6 with citric acid monohydrate.
[0070] In one embodiment, the amount of enzyme is in the range of 0.2%, 0.5% to 10% of the weight of the plant material. In another embodiment, the pH of the mixture is 3-10. In a particular embodiment, the enzyme concentration and pH level of the mixture produce optimal enzyme activity. In one embodiment, the lipid is one or more lipids selected independently from the group consisting of olive oil, coconut oil, sesame oil, vegetable oil, milk, butter, liposomes and hemp seed oil and / or other vegetable solvents and / or food, such as glycerin, polyethylene glycol, ethyl acetate, d-limonene, butylene glycol, propylene glycol, ethylhexyl palmitate and / or with the addition of lecithin. In one embodiment, the lipid is olive oil. In another embodiment, the lipid is coconut oil. In another embodiment, the lipid is sesame oil. In another embodiment, the lipid is vegetable oil. In yet another embodiment, the lipid is milk. In another embodiment, the lipid is butter.
[0071] In one embodiment, the weight ratio of lipid to plant material is in the range of 0.01: 1 to 4: 1 and the weight ratio of water to plant material is in the range of 0.01: 1 to 10: 1. In another embodiment, the weight ratio of lipid to material
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25/52 plant is in the range of 0.1: 1 to 2: 1 and the weight ratio of water to plant material is in the range of 1: 1 to 5: 1. In a particular embodiment, the weight ratio of lipid to plant material is in the range of 0.5: 1 to 1.5: 1 and the weight ratio of water to plant material is in the range of 2: 1 to 3 :1. The weight ratio of the lipid to the plant material is preferably in the range of 2: 3 and the weight ratio of water to plant material in the dry matter is in the range of 0.01: 1 to 10: 1, preferably in the range of 2: 1.
[0072] In one embodiment, the mixture is treated with ultrasound before adding the enzymes. In one embodiment, the mixture is microwaved before adding the enzymes.
[0073] In one embodiment, the mixture is treated with ultrasound after adding the enzymes. In one embodiment, the mixture is treated with a microwave after adding the enzymes.
[0074] In one embodiment, lipids, water and enzymes are added in any different order combinations.
[0075] In a particular embodiment, the displacement of the vegetable matter, adding the lipids, adding the water and adding the enzymes is done in any combination of different order.
[0076] In one embodiment, the mixture is stirred for at least 10 minutes, preferably 30 or 60 minutes.
[0077] In one embodiment, the mixture is stirred at a temperature between 40 and 70 ° C.
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26/52 [0078] In one embodiment, the mixture is separated by density. In another embodiment, the mixture is separated by pressing and / or filtering.
[0079] In another embodiment, the mixture is separated into a lipid phase and a wet solid phase. In one embodiment, the fat-soluble extract is recirculated any number of times to achieve a higher cannabinoid or terpene content.
[0080] In one embodiment, the fat-soluble extract is recirculated any number of times to achieve greater stability for the cannabinoid or terpene.
[0081] In another embodiment, at least 50%, preferably 70% of the terpenoids, at least 70% of the diterpenoids and at least 50%, preferably 70% of the monoterpenes contained in the plant material are extracted into the fat-soluble extract.
[0082] In yet another embodiment, at least 70% of the sesquiterpenes and at least 50% of the monoterpenes contained in the plant material are extracted for the fat-soluble extract.
[0083] In one embodiment, the fat-soluble extract has a total cannabinoid content of at least 2 weight percent. In another embodiment, the lipid-based extract has a total cannabinoid content of at least 3 weight percent. In yet another embodiment, the lipid-based extract has a total cannabinoid content of at least 5 weight percent.
[0084] In one embodiment, the two main cannabinoids in the fat-soluble extract are
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27/52 preferably, THC and CBD, or any other cannabinoids.
[0085] In another embodiment, the invention provides a fat-soluble extract obtained from the process of producing a fat-soluble extract containing cannabinoids and terpenoids from plant material, chosen from the group consisting of hemp, cannabis, hops, echinacea, sage dinivorum, chrysanthemum , helichrysum and hypericum biomass and plant material from Tanacetum cinerariifolium, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with enzymes to form a mixture to which water and lipids or solvents can be optionally added;
ç. stirring the mixture at a temperature between 1 and 80 ° C; and
d. separating the mixture into a lipid phase, an aqueous phase and a solid phase;
wherein the lipid phase comprises the fat-soluble extract.
[0086] The extract is surprisingly stable 60 days after extraction.
[0087] Preferably, in fat-soluble extract, the degradation of cannabinoids after four weeks in the dark at 25 ° C is less than 5% and has a total cannabinoid content of at least 90%, preferably at least 95%, the ratio between the two main cannabinoids in the fat-soluble extract differs by less than 10%, preferably less than 5%, the ratio between the two main cannabinoids in plant material and less than 10%, preferably less
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28/52 of 5%, more preferably less than 2%, of cannabinoids are decarboxylated during the process, and the monoterpene / diterpene ratio after four weeks in the dark at 25 ° C is less than 5%, and the monoterpene content is at 80% of the initial content.
[0088] In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, in which the fingerprint of plant material is reproduced in the extract, meaning that the variation in the proportion of the two main cannabinoids compared to the starting plant material ratio is less than 10%.
[0089] In another embodiment, the THCtot: CBDtot ratio in the fat-soluble extract differs by less than 5% from THCtot: CBDtot in plant material.
[0090] In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, in which the decarboxylated forms of cannabinoids represent less than 10% of the total cannabinoid content.
[0091] In yet another aspect, this document provides the stability of cannabinoids in the lipid phase which is significantly increased by maintaining at least 90% of the cannabinoid content after two weeks at room temperature and in the dark.
[0092] In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, in which the extract has a content of terpenes / terpenoids of at least 75% of the initial content of material vegetable by weight.
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29/52 [0093] In yet another aspect, the present document provides a fat-soluble extract that can be obtained from the process according to the invention, in which the monoterpene content is at least 30% of the total terpene content.
[0094] The fat-soluble extract according to the invention has surprisingly a cannabinoid content of:
• DELTA-9-TETRAIDROCANABINOL (THC-ACID): in a range of 1,000 to 6,000 mg / kg • DELTA-9-TETRAIDROCANABINOL (THC-NEUTRAL): in a range of 100 to 500 mg / kg • DELTA-9-TETRAIDROCANABINOL (THC-TOTAL EXPRESSED AS NEUTRAL THC): in the range of 1,000 to 7,000 mg / kg • CANABIDIOL (CBD): in the range of 1,000 to 5,000 mg / kg • CANABIDIOL ACID (CBD-A): in the range of 20,000 to 80,000 mg / kg.
[0095] In one embodiment, the plant material is heated before forming the mixture for the decarboxylation of cannabinoids. In one embodiment, the mixture is heated for the decarboxylation of cannabinoids. In one embodiment, the lipid extract is heated for the decarboxylation of cannabinoids.
[0096] In one embodiment, the cannabinoid content of the solid phase is reduced by at least 75 weight percent. In another embodiment, the cannabinoid content of the solid phase is reduced by at least 80 weight percent. In another embodiment, the cannabinoid content of the solid phase is reduced by at least 90 weight percent.
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30/52 [0097] In one embodiment, the solid phase and the fat-soluble extract are used for the formulation of food and feed products.
[0098] In another embodiment, the aqueous phase and the lipid-soluble extract are used in the production of the pharmaceutical substance, nutraceutical products, cosmetics, food or feed, antimicrobial, antibacterial products, insecticides or biocides.
[0099] In another aspect, this document provides a fat-soluble extract, in which the extract has a total phyto-cannabinoid content of at least 5 percent by weight.
[00100] In one embodiment, the plant material includes seeds. In another embodiment, when the plant material includes seeds, no lipid is added. In another embodiment, when the plant material includes seeds, a lipid is added.
[00101] In one embodiment, the plant material is a mixture comprising buds, flowers, stems, leaves, roots and seeds. In another embodiment, when the plant material is a mixture comprising buds, flowers, stems, leaves, roots and seeds, a lipid is added to obtain an optimal proportion of lipid-to-plant material for the effective extraction of cannabinoids. In another embodiment, when the plant material is a mixture comprising buds / flowers and seeds, a lipid is not added.
[00102] In one embodiment, the plant material is freshly harvested and contains a high level of humidity; in that case, adding extra water to the plant material is unnecessary.
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31/52 [00103] In one embodiment, the plant material has a lipid content of at least 1 weight percent.
[00104] In another embodiment, the addition of lipids reaches a lipid content of 5 weight percent of the mixture.
[00105] In one embodiment, the mixture is stirred for at least 10 minutes.
[00106] In one embodiment, the mixture is separated by density. In another embodiment, the mixture is separated by pressing and filtering.
[00107] In one embodiment, the mixture is stirred at a temperature between 40 and 70 ° C.
[00108] In one embodiment, the plant material is hemp comprising less than 0.6% THCtot. In yet another embodiment, the plant material is marijuana comprising more than 0.6% THCtot. In another embodiment, the plant material is a hybrid or genetically modified variant of hemp. In yet another embodiment, the plant material is a hybrid or genetically modified variant of cannabis.
[00109] In one embodiment, the enzyme is one or more enzymes selected independently from the group consisting of cellulase, beta-glucosidase, hemicellulase, xylanase, glucanase, pectinase, amylase, phospholipase, betamananase, arabinanase, phytase and protease. In one embodiment, the enzyme is cellulase. In another embodiment, the enzyme is beta-glycosidase. In another embodiment, the enzyme is hemicellulase. In another embodiment, the enzyme is xylanase. In yet another form of
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32/52
achievement, The enzyme is glucanase. Still in another form in achievement, The enzyme is pectinase. Still in another form in achievement, The enzyme is amylase. In yet another form in achievement, The enzyme is phospholipase. Still in another modality, The enzyme is beta-mannanase. Still in another way
the enzyme is arabinanase. In yet another embodiment, the enzyme is phytase. In another embodiment, the enzyme is protease.
[00110] In one embodiment, the amount of enzyme is 0.5% to 10%, preferably 3%, of the weight of the plant material. In another embodiment, the pH of the mixture is 3-10, preferably pH 5.6. In a particular embodiment, the enzyme concentration and pH level of the mixture produce optimal enzyme activity.
[00111] In one embodiment, the lipid is one or more lipids selected independently from the group consisting of olive oil, coconut oil, sesame oil, vegetable oil, milk, butter, liposomes, hemp seed oil and / or others vegetable and / or food solvents such as glycerin, polyethylene glycol, ethyl acetate. In one embodiment, the lipid is olive oil. In another embodiment, the lipid is coconut oil. In another embodiment, the lipid is vegetable oil. In yet another embodiment, the lipid is milk. In another embodiment, the lipid is butter.
[00112] In one embodiment, the weight ratio of lipid to plant material is in the range of 0.01: 1 to 4: 1 and the weight ratio of water to plant material is in the range of 0.01: 1 to 10: 1. In another form of
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In this embodiment, the weight ratio of lipid to plant material is in the range of 0.1: 1 to 2: 1 and the weight ratio of water to plant material is in the range of 1: 1 to 5: 1. In a particular embodiment, the weight ratio of lipid to plant material is in the range of 0.5: 1 to 1: 1.5 and the weight ratio of water to plant material is in the range of 2: 1 to 3 :1.
[00113] In one embodiment, the mixture is treated with ultrasound before adding the enzymes. In one embodiment, the mixture is microwaved before adding the enzymes.
[00114] In one embodiment, the mixture is treated with ultrasound after adding the enzymes. In one embodiment, the mixture is treated with a microwave after adding the enzymes.
[00115] In one embodiment, lipids, water and enzymes are added in any different order combinations.
[00116] In a particular embodiment, the displacement of plant matter, adding lipids, adding water and adding enzymes is done in any combination of different order.
[00117] In one embodiment, the fat-soluble extract is recirculated any number of times to achieve a higher phyto-cannabinoid content.
[00118] In one embodiment, the fat-soluble extract is recirculated any number of times to achieve superior phyto-cannabinoid stability.
[00119] In one embodiment, the fat-soluble extract has a total phyto-cannabinoid content of at least
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34/52 minus 2 weight percent. In another embodiment, the lipid-based extract has a total cannabinoid content of at least 3 weight percent. In yet another embodiment, the lipid-based extract has a total cannabinoid content of at least 5 weight percent.
[00120] In A way of achievement, the material vegetable is heated before forming the mixture to The decarboxylation in cannabinoids. In a form in the mixture is heated for decarboxylation in cannabinoids. In A way of achievement, the extract fat-soluble is heated to decarboxylation in cannabinoids.[00121] In A way of achievement, the content in phyto-cannabinoids gives solid phase is reduced by at least
percent by weight. In another embodiment, the phyto-cannabinoid content of the solid phase is reduced by at least 80 weight percent. In another embodiment, the phyto-cannabinoid content of the solid phase is reduced by at least 90 weight percent.
[00122] In one embodiment, the solid phase is used for the formulation of food products and feed.
[00123] In another embodiment, the aqueous phase can be used to obtain nutraceutical, antimicrobial, antibacterial or biopesticidal products.
[00124] In yet another aspect, this document provides a solid extract of cannabis or hemp plant material, in which the cannabinoid content of
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35/52 plant material is reduced by at least 90% by weight.
[00125] In one embodiment, in which the solid phase or extract is produced by a process to produce a liquid-soluble extract containing cannabinoids and terpenes of plant material, chosen from the group consisting of hemp, cannabis, hops, echinacea, salvia dinivorum, chrysanthemum, helichrysum and hypericum biomass, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with water and lipids to form a mixture;
ç. stirring the mixture at a temperature between 1 and 80 ° C; and
d. separating the mixture into a lipid phase, an aqueous phase and a solid phase;
wherein the lipid phase comprises the fat-soluble extract.
[00126] In one embodiment, the plant material includes seeds. In another embodiment, when the plant material includes seeds, no lipid is added. In another embodiment, when the plant material includes seeds, a lipid is added.
[00127] In one embodiment, the plant material is a mixture comprising buds, flowers, stems, leaves, roots and seeds. In another embodiment, when the plant material is a mixture comprising buds, flowers, stems, leaves, roots and seeds, a lipid is added to obtain an optimal proportion of lipid-to-plant material for the effective extraction of cannabinoids. In another embodiment, when the
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36/52 plant material is a mixture comprising seeds and buds, flowers, stems, leaves, roots and seeds, a lipid is not added.
[00128] On an form in achievement, the material vegetable has a content in seeds less than 98% the weight of material vegetable and O material vegetable (different from) seeds is superior to 2% of the weight of plant material.[00129] On an form in achievement, the material
vegetable has a lipid content of at least 1 weight percent of plant material.
[00130 ] In another embodiment, The addition in lipids reaches one 5 percent lipid content by weight gives mixture. [00131 ] In an embodiment, The mixture is agitated during fur minus 10, preferably 30 minutes •[00132 ] In an embodiment, The mixture is
separated by density. In another embodiment, the mixture is separated by pressing and filtering.
[00133] In one embodiment, the mixture is stirred at a temperature between 40 to 75 ° C, preferably 55 ° C.
[00134] In one embodiment, the plant material is hemp comprising less than 0.6% THC. In yet another embodiment, the plant material is cannabis comprising more than 0.6% THC. In another embodiment, the plant material is a hybrid or genetically modified variant of hemp. In yet another embodiment, the plant material is a hybrid or genetically modified variant of cannabis.
[00135] In one embodiment, the enzyme is
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37/52 one or more enzymes selected independently from the group consisting of cellulase, beta-glucosidase, hemicellulase, xylanase, glucanase, pectinase, amylase, phospholipase, betamananase, arabinanase, phytase and protease. In one embodiment, the enzyme is cellulose. In yet another embodiment, the enzyme is beta-glycosidase. In another form of the enzyme is hemicellulase. In another embodiment, the enzyme is xylanase. In yet another embodiment, the enzyme is glucanase. In yet another embodiment, the enzyme is pectinase. In yet another embodiment, the enzyme is amylase. In yet another embodiment, the enzyme is phospholipase. In yet another embodiment, the enzyme is beta-mannanase. In yet another embodiment, the enzyme is arabinanase. In yet another embodiment, the enzyme is phytase. In another embodiment, the enzyme is protease.
[00136] In one embodiment, the amount of enzyme is 0.5% to 10% of the weight of the plant material. In another embodiment, the pH of the mixture is 3-10. In a particular embodiment, the enzyme concentration and the pH level of the mixture produce optimal enzyme activity.
[00137] In one embodiment, the lipid is one or more lipids selected independently from the group consisting of olive oil, coconut oil, vegetable oil, milk, hemp seed oil and butter. In one embodiment, the lipid is olive oil. In another embodiment, the lipid is coconut oil. In another embodiment, the lipid is vegetable oil. In yet another embodiment, the lipid is milk. In yet another form of
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38/52 realization, the lipid is glycerin. In another embodiment, the lipid is butter.
[00138] In one embodiment, the weight ratio of lipid to plant material is in the range of 0.01: 1 to 4: 1 and the weight ratio of water to plant material is in the range of 0.01: 1 to 10: 1. In another embodiment, the weight ratio of lipid to plant material is in the range of 0.1: 1 to 2: 1 and the weight ratio of water to plant material is in the range of 1: 1 to 5: 1. In a particular embodiment, the weight ratio of lipid to plant material is in the range of 0.5: 1 to 1: 1.5 and the weight ratio of water to plant material is in the range of 2: 1 to 3 :1.
[00139] In one embodiment, the mixture is treated with ultrasound before adding the enzymes. In one embodiment, the mixture is microwaved before adding the enzymes.
[00140] In one embodiment, the mixture is treated with ultrasound after adding the enzymes. In one embodiment, the mixture is treated with a microwave after adding the enzymes.
[00141] In one embodiment, lipids, water and enzymes are added in any different order combinations.
[00142] In a particular embodiment, the grinding of plant matter, addition of lipids, addition of water and addition of enzymes are carried out in any combination of different order.
[00143] In one embodiment, the fat-soluble extract is recirculated any number of times to
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39/52 achieve a higher cannabinoid content.
[00144] In one embodiment, the fat-soluble extract has a total cannabinoid content of at least 2 weight percent. In another embodiment, the lipid-based extract has a total cannabinoid content of at least 3 weight percent. In yet another embodiment, the lipid-based extract has a total cannabinoid content of at least 5 weight percent.
[00145] In one embodiment, the plant material is heated before forming the mixture for the decarboxylation of cannabinoids. In one embodiment, the mixture is heated for the decarboxylation of cannabinoids. In one embodiment, the fat-soluble is heated for the decarboxylation of cannabinoids.
[00146] In one embodiment, the cannabinoid content of the solid phase that can be obtained from the process according to the present invention is reduced by at least 75% by weight. In another embodiment, the cannabinoid content of the solid phase is reduced by at least 80 weight percent. In another embodiment, the cannabinoid content of the solid phase is reduced by at least 90 weight percent.
[00147] In one embodiment, the solid phase is used for the formulation of food products and feed.
[00148] In another embodiment, the aqueous phase can be used in the production of nutraceuticals, antimicrobials, antibacterials or biopesticides.
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40/52 [00149] In yet another aspect, the present document provides that it is the use of the solid extract or phase of cannabis or hemp plant material that can be obtained from the process according to the present invention for the formulation of foods or rations.
[00150] In another aspect, the aqueous phase can be used in the production of nutraceuticals, antimicrobials, antibacterials or biopesticides.
[00151] In yet another aspect, this document provides the use of fat-soluble extract for the preparation of a cream or gel containing at least 0.5% cannabinoids, showing an increase in the stability of cannabinoids by at least 90% of the initial content after 10 weeks.
[00152] In yet another aspect, this document provides for the use of fat-soluble extract for the preparation of a gum or candy containing at least 0.5% cannabinoids exhibiting an increased cannabinoid stability of at least 90% of the initial content after 10 weeks.
[00153] In yet another aspect, this document provides the use of fat-soluble extract for the preparation of a gel containing at least 0.5% cannabinoids exhibiting an increased cannabinoid stability of at least 90% of the initial content after 10 weeks.
[00154] In yet another aspect, the use of the aqueous phase can be obtained from the process according to the invention, for the preparation of pharmaceutical or nutraceutical products, cosmetics, food products or
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41/52 food products, antimicrobials, antibacterials, insecticides or biopesticides.
EXAMPLES [00155] The invention is now described with reference to the following Examples. These Examples are provided for illustrative purposes only, and the invention is not limited to these Examples, but covers all variations that are evident as a result of the teachings provided in this document.
Example 1:
[00156] A quantity of 150 g of dry industrial hemp buds, comprising few seeds, variety Futura 75 were mixed in a domestic kitchen mixer Mulinex Companion with 340 g of water, 100 g of extra virgin olive oil offered by Azienda Agrícola Montesoli, Siena . The temperature of the mixture was raised and maintained at 55 ° C with constant stirring at 100 rpm for 3.5 h. The mixture was then centrifuged at 11,000 rpm for 10 min. No lipid extract could be obtained, as the lipids remained adsorbed on the plant matrix. The process was then repeated with the only difference that a cocktail of commercial food-grade enzymes was added to the mixture and the pH adjusted to pH 5.6 with 6 g of citric acid monohydrate. The enzyme cocktail comprised Celluclast 1.5 L (cellulase), Ultraflow Max (betaglucanase), Peclyve (pectinase, beta-glucanases, cellulases and beta-mannanases) and Ceremix 2XL (alpha-amylase, beta-glucanase, protease). The total enzyme concentration was 3% by weight of the material of the hemp plant. After 3.5 h the mixture became homogeneous. After centrifuging the
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42/52 mixture (11,000 rpm for 5 min.), 91 g of fat-soluble extract, 80 g of an intermediate aqueous phase and 410 g of a wet solid fraction were recovered. The solid fraction was dried in an oven at 50 ° C for 6 h. Futura 75 hemp buds and lipid extract were sent for analysis of cannabinoids and terpenes in an accredited laboratory. The solid fraction was analyzed only for cannabinoid content.
[00157] The methodology used for the analysis of cannabinoids is UPLC-MS / MS, with detection limit for di THC and THC acid not less than 1.0 mg / kg in oil and 0.10 mg / kg in hemp flour and seeds. The Δ-9tetrahydrocannabinol and its acid derivative were extracted with a mixture of methanol and dichloromethane for the solid material or another mixture based on methanol for the oil. Chromatographic conditions: phase A: water + 0.1% (v / v) formic acid, phase B: acetonitrile + 0% (v / v) formic acid. Flow: 0.5 mL / min., Column: .Waters® UPLC BEH C18 2.1 x 100 mm, 1.7 pm or equivalent. Column temperature: 35 ° C. Temperature self-sampling: 8 ° C. Conditions
mass Spectrometer: Source in temperature: 130 ° C. Temperature desolventization: 400 ° Ç. Capillary: 1 KV. Flow: 1,000 L / h. Cone Flow: 50 L / h. . 0 total of THC is calculated from according to the following formula:
C tot = Ca x PMn / PMa = Cn
C tot = analyte content (total THC) in the sample analyzed, in mg / kg
Cn = calculated THC neutrality concentration, in mg / kg
Ca = THC acid concentration calculated in mg / kg
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43/52
PMa = molecular weight THC acid
PMn = neutral molecular weight THC
The following concentrations of cannabinoids (mg / kg) were found on the buttons:
• DELTA-9-TETRA-HIDR0CANABIN0L (THC-ACID): 1,229 • DELTA-9-TETRAIDROCANABINOL (THC-NEUTRAL): 142,8 • CANABIDIOL (CBD): 2,328 • CANABIDIOL ACID (CBD-A): 21,230 • CANABINOL ( CBN): 10 [00158] The cannabinoid content in the lipid fraction was as follows (mg / kg):
• DELTA-9-TETRAHYDROCANABINOL (THC-ACID): 1,983 • DELTA-9-TETRAHYDROCANABINOL (THC-NEUTRAL): 146.8 • CANABIDIOL (CBD): 3,750 • CANABIDIOL ACID (CBD-A): 23,246 • CANABINOL (CBN): 12 [00159] Considering the relationship between the oil-plant material (2 to 3), an efficiency for THC (TOTAL) of 2, 130 * (2/3) * 0, 91 / 1,372 = 94.2% was achieved. As can be seen, a CBD content of 2.3%, even higher than the initial materials of the plant, was achieved. Considering the rest of the test mass, a 93.8% cannabinoid extraction yield was obtained, using 15 times less solvent than alternative scientific lipid based extraction methodologies (Romano and Hazekamp, 2013; Cannazza, 2016) . In addition, the process was able to reproduce the fingerprint of the plant material in the extract, keeping the variation in the ratio between the two main cannabinoids CBD and THC below 5%. The THCtot: CBDtot ratio in plant material and extract was 0.058
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44/52 and 0.079, respectively. The variation in the ratio was therefore limited to 0.02 or 2%. Furthermore, the process did not decarboxylate or degrade cannabinoids as the ratio of CBDA: CBD and THCA: THC was not altered in the plant material and in the lipid extract, as well as the low content of CBN demonstrates.
[00160] The solid fraction had a cannabinoid content as follows (mg / kg):
• DELTA-9-TETRAIDROCANABINOL (THC-ACID): 21 • DELTA-9-TETRAIDROCANABINOL (THC-NEUTRAL): 9 • CANABIDIOL (CBD): 390 • CANABIDIOL ACID (CBD-A): 5,986 [00161] As can be seen , a significant reduction in THC in plant material was achieved.
[00162] The lipid extract was also analyzed for digital printing of terpene. GC-MS analyzes were performed in an Agilent 5973N selective mass detector coupled to an Agilent 6890 gas chromatograph (Paio Alto, CA), equipped with an HP5-MS capillary column (30 mx 0.25 mm x 0.25 qm) , operating in 70eV electronic ionization mode, with transfer line maintained at 260 ° C, while quadrupole and ion source temperature were maintained at 150 ° C and 230 ° C, respectively. Helium (1.0 mL min ^-1 ) was used as the carrier gas. The injector temperature was maintained at 250 ° c and the oven temperature program was from 60 ° C to 240 ° c at a rate of 3 min ^-1 . The detector (FID) was operated at 280 ° C. The lipid extract (0.03 μΕ) was injected in split mode (100: 1). A standard solution of n-alkanes (C7 - C26) was used to obtain the retention indices. Individual volatile components have been identified by
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45/52 comparison of their mass spectra (MS) and retention indices (IR) with those reported in the literature.
[00163] The main terpenes contained in the lipid extract were as follows:
Compound Percentage Alpha-Pineno 14.4 Beta-Pinene 5, 3 Beta-Myrcene 21.4 Limonene 5, 2 Terpinolene 10.5 Beta-Karyophylene 17, 1 Beta-Humulene 6, 1
[00164] The terpene fingerprint of the obtained extract is very comparable to that reported by Nissen et al. (2010). Taking into account that, in our case, the initial plant material was dried and was not harvested recently, it can be noted that no significant variation of the original fingerprint nor the degradation of cannabinoids and terpenes occurred during the process.
Example 2:
[00165] A quantity of 5 kg of hemp seeds (oil content of 34%) was processed with a Bracco Sri mechanical spindle type expeller, Italy, 7.5 kW, 50 kg / h. The extracted oil and the seed cake were analyzed for CBD-A content. The oil extracted with a mechanical expeller had a CBD-A content of 16.3 mg / Kg, while the seed cake had a CBD-A content of
17.8 mg / kg.
[00166] A quantity of 200 g of seeds was
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46/52 processed using the enzyme-assisted extraction protocol of Example 1 with the only difference that no external oil source was added as the oil was already included in the seeds. The oil extract and the solid fraction were collected. The solid fraction was kiln dried at 50 degrees Celsius. The oil and the solid fraction were sent for CBD-A analysis. The oil extracted with our protocol aided by enzymes had a CBD-A content of 21.1 mg / kg. The dry solid fraction had a CBD-A content of 4.3 mg / kg.
[00167] As can be seen, the proposed method increased the cannabinoid content in the lipid fraction, while significantly reducing the cannabinoid content in the residual protein-rich solid fraction.
Example 3:
[00168] A quantity of 100 g of hemp-based lipid extract prepared with the process described in the previous Example, having a CBD-A content of 21.3, was added to the Futura 75 hemp buds in replacement of olive oil using the same protocol of Example 1. Without enzymes, no lipid extract was recovered. With enzymes, 92 grams of lipid extract were recovered.
[00169] The cannabinoid content of the recovered lipid extract was (mg / kg):
• DELTA-9-TETRAHYDROCANABINOL (THC-ACID): 1,440 • DELTA-9-TETRAHYDROCANABINOL (THC-NEUTRAL): 133.2 • CANNABIDIOL (CBD): 3,819 • Acid CANNABIDIOL (CBD-A): 25,388 [00170] How can you be seen, using an enzymatic approach to produce a hemp seed oil and
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47/52
then using the same oil to extract sprouts in hemp, the content of CBD and CBD-A reachable in extract is even higher than the initial hemp. Example 4: [00171] 0 Example 1 was repeated with one sonication treatment of the mixture before addition in
enzymes. Treatment conditions: 250 W ultrasonic power, 30 min. ultrasonic time and 50 ° C ultrasonic temperature. It was found that the ultrasonic treatment allowed a reduction of the time from 3.5 hours to 2.5 to achieve the comparable cannabinoid extraction yield described in Example 1.
Example 5:
[00172] Analysis of the cannabinoid content in the lipid fraction of example 1 was repeated after 21 days and after 60 days. After 21 days, the cannabinoid content was:
• DELTA-9-TETRAHYDROCANABINOL (THC-Acid): 1,935 • DELTA-9-TETRAHYDROCANABINOL (THC-NEUTRAL): 148.9 • CANNABIDIOL (CBD): 3,754 • CANABIDIOL ACID (CBD-A): 23,238 [00173] After 60 days, the cannabinoid content was:
• DELTA-9-TETRAHYDROCANABINOL (THC-ACID): 1,937 • DELTA-9-TETRAHYDROCANABINOL (THC-NEUTRAL): 149.8 • CANNABIDIOL (CBD): 3,757 • CANABIDIOL ACID (CBD-A): 23,216 [00174] In both In cases, no significant reduction in cannabinoid content was found.
Example 6:
[00175] A quantity of 375 g of fresh buds recently harvested from Futura 75, having a content of
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48/52 humidity of 60%, was treated as in example 1 with the only difference that no water was added to the mixture, since the initial humidity was sufficient for enzymatic activity. The same amount of lipid extract was obtained (91 g). The lipid extract was sent for analysis of cannabinoids and terpenes.
[00176] Cannabinoid content (mg / kg):
• DELTA-9-TETRAIDROCANABINOL (THC-Acid): 2,130 • DELTA-9-TETRAIDROCANABINOL (THC-NEUTRAL): 126,2 • CANNABIDIOL (CBD): 1,950 • CANNABIDIOL ACID (CBD-A): 25,347 Terpene fingerprints the following:
Compound Percentage Alpha-Pineno 16.4 Beta-Pinene 6, 3 Beta-Myrcene 20.4 Limonene 5, 3 Terpinolene 10.4 Beta-Karyophylene 16.1 Alpha-Humulene 5, 9
[00177] In this way, it was possible to obtain not only a high yield of cannabinoid extraction, but also the extraction of the entire fingerprint of the terpene, including volatile monoterpenes.
Example 7:
[00178] A quantity of 150 g of dry roots of Echinacea purpurea (purple coneflora) was mixed in a Mulinex Companion kitchen mixer with 340 g of water, 100 g of extra virgin olive oil offered by Azienda Agrícola Montesoli, Siena and a cocktail of enzymes
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49/52 food quality commercials Celluclast 1.5 L (cellulase), Ultraflow Max (betaglucanase), Peclyve (pectinase, beta-glucanases, cellulases and beta-mannanases) and Ceremix 2XL (alpha-amylase, beta-glucanase, protease). The total concentration of enzymes was 3% of the weight of the Echinacea plant. The pH of the mixture was adjusted to pH 5.6 with 6 g of citric acid monohydrate, while the temperature of the mixture was raised and maintained to 55 with constant stirring at 100 rpm. After 3.5 h the mixture is homogeneous. After centrifuging the mixture (11,000 rpm for 5 min.), 92 g of fat-soluble extract, 80 g of an intermediate aqueous phase and 409 g of a wet solid fraction were recovered. The lipid fraction was sent for analysis of the total content of N-alkylamides (N-alkamides) by HPLC which consisted of a Beckman System Gold 126 solvent module, a Beckman model 508 autosampler, a Beckman model 168 detector (Beckman Coulter, Inc. , Fullerton, CA), and an identification of 250 x 10 mm, 5 pm ODC-AM-303 RP-ColumnCis (YMC, Inc., Wilmington, NC). The HPLC method for analysis
of alcamide was the same as reported by Senchina and others . [00179] The mobile phases for the gradient in alcamide were the following: (THE) Milli water -Q degassed and (B) acetonitrile. A linear gradient in
an increase of 40% B to 80% B was developed within 45 min. at a flow rate of 1.0 mL / min. with UV detection from 200 to 600 nm. The injection volume was 15 pL. All Echinacea extracts were filtered through 0.45 pm polytetrafluoroethylene filters (Alltech Associates Inc., Deerfield, IL) before being injected on the HPLC. The contents
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50/52 total N-alkylamides in the roots and lipid fractions were as follows (mg / kg):
• Roots: 3.016 mg / Kg • Lipid extract: 3.257 mg / kg [00180] After 4 weeks at room temperature, the analysis of N-alkylamides was repeated for the fat-soluble extract with the following result:
• 3.197 mg / kg [00181] As can be seen, the variation in the content of N-alkylamides in the lipid-based extract was very limited.
Example 8:
[00182] A quantity of 150 g of dried flowers of cinerariaefolium chrysanthemum (1.2% content) of pyrethrin I and pyrethrin II were purchased from local farms in Kenya and mixed in a domestic auxiliary mixer Mulinex Companion with 340 g of water, 100 g of extra-virgin olive oil and a cocktail of commercial food-grade enzymes including Celluclast 1.5 L (cellulase), Ultraflow Max (betaglucanase), Peclyve (pectinase, beta-glucanases, cellulases and beta-mannanases) and Ceremix 2XL (alpha-amylase , beta-glucanase, protease). The total enzyme concentration was 3% by weight of the chrysanthemum plant material. The pH of the mixture was adjusted to pH 5.6 with 6 g of citric acid monohydrate, while the temperature of the mixture was raised and maintained at 55 ° C with constant stirring at 100 rpm. After 3 hours the mixture is homogeneous. After centrifuging the mixture (11,000 rpm for 5 min.), 94 g of fat-soluble extract, 81 g of an intermediate aqueous phase and 405 g
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51/52 of a wet solid fraction. The initial plant material and lipid extract were sent for analysis of the Pyrethrin content by HPLC using a Beckman System Gold 126 solvent module, a Beckman model 508 automatic sampler, a Beckman model 168 detector (Beckman Coulter, Inc., Fullerton, CA ) and an id 250 x 10 mm, 5 pm ODC-AM-303 RP-Column C18 (YMC, Inc., Wilmington, NC). The HPLC method for pyrethrin analysis was the same proposed by Marr. The elution was carried out with a mixture of acetyl acetate and hexane in a proportion of 1:10 at a constant flow rate of 1.5 ml per minute, leading to an analysis of 15 min. The UV detector was adjusted to a wavelength of 242 nm. A refined pyrethrin sample was purchased from RdH laborchemikalien & Co. KG (Germany) for standardization of the analytical method. Its pyrethrin content was claimed to be 21.1%.
[00183] The total pyrethrin content (pyrethrin I and II) was 1.4%. Pyrethrin I to Pyrethrin II ratio was 1.65. After 4 weeks of darkness at room temperature, the analysis for pyrethrins was repeated for the fat-soluble extract. The pyrethrin content was 1.37% mg / kg, while the pyrethrin I to pyrethrin II ratio was 1.68.
[00184] As can be seen, not only was the extraction very efficient, but the stability of the pyrethrins in the lipid extract was also significant.
[00185] Although the invention has been disclosed with reference to specific embodiments, it is evident that other embodiments and variations of this invention can be designed by others skilled in the art without departing from the true spirit and scope of the invention. At
Petition 870190065723, of 12/07/2019, p. 58/79
52/52 attached claims are intended to be interpreted to include all such embodiments and equivalent variations.

权利要求:
Claims (57)
[1]
1. Process for the production of a fat-soluble extract from plant material containing phytocannabinoids and / or terpenoids and / or terpenes, comprising the steps of:
The. crushing plant material;
B. mixing the crushed plant material with water and lipids to form a mixture to which water and lipids are optionally added;
ç. stirring the mixture at a temperature between 1 and 80 ° C; and
d. separating the mixture into a lipid phase, an aqueous phase and a solid phase;
wherein the lipid phase comprises the fat-soluble extract.
[2]
A process according to claim 1, wherein the lipids are added to said mixing step b.
[3]
Process according to claim 1, in which water is added to said mixing step b.
[4]
Process according to any one of claims 1 to 3, wherein said plant material is chosen from the group consisting of buds, flowers, leaves, stems, roots and seeds or a mixture thereof.
[5]
Process according to any one of claims 1 to 4, wherein said plant material containing phyto-cannabinoids or terpenoids is chosen from the group consisting of hemp, cannabis, hops, echinacea, sage dinivorum, chrysanthemum, helichrysum and hypericum biomass. and in which said plants are pure hybrids
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2/10 or genetically modified.
[6]
Process according to any one of claims 1 to 5, wherein said plant material is derived from the genus Cannabis of plants, which comprises the species C. sativa, C. indica and C. ruderalis.
[7]
Process according to any one of claims 1 to 6, wherein said plant material is industrial hemp of the species C. sativa.
[8]
Process according to any one of claims 1 to 7, wherein said enzymes from step b. are one or more enzymes selected independently from the group consisting of cellulase, hemicellulase, xylanase, glucanase, beta-glucanase, pectinase, amylase, alpha-amylase, beta-amylase, phospholipase, arabanase, galacto-, beta-mannanase, protease and phytase.
[9]
Process according to any one of claims 1 to 8, wherein the plant material has a moisture content of at least 20% by weight of the plant material.
[10]
A process according to claims 1 to 9, wherein said plant material is collected recently and has a moisture content of at least 30%, preferably at least 40%.
[11]
A process according to any one of claims 1 to 10, wherein the plant material has a total phyto-cannabinoid content greater than 0.2% of the weight of the plant material.
[12]
Process according to any one of claims 1 to 11, in which the plant material is industrial hemp comprising less than 0.2-0.6% THC, or in which the plant material is cannabis comprising more than
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3/10 than 0.2-0.6% THC, or hybrids and their genetically modified variants.
[13]
Process according to any one of claims 1 to 12, wherein the plant material has a seed content of less than 98% of the weight of the plant material.
[14]
A process according to any one of claims 1 to 13, wherein the plant material includes a mixture of cannabis and other plants; in which in such a mixture the phyto-cannabinoid content is greater than 2%.
[15]
Process according to any one of claims 1 to 14, wherein the plant material other than seeds is greater than 2% of the weight of the plant material.
[16]
A process according to any one of claims 1 to 15, wherein the plant material has a lipid content greater than 1% by weight of the plant material.
[17]
A process according to any one of claims 1 to 16, wherein the addition of lipids reaches a lipid content of at least 5% by weight of the mixture.
[18]
A process according to any one of claims 1 to 17, wherein the mixture is stirred for at least 10 minutes.
[19]
Process according to any one of claims 1 to 18, wherein the mixture is stirred at a temperature of 40 to 75 ° C.
[20]
A process according to any one of claims 1 to 19, wherein the mixture is separated by density or by pressure and / or filtration of the mixture.
[21]
21. Process according to any one of claims 1 to 20, wherein the mixture is separated into a
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4/10 lipid phase and a solid hybrid phase.
[22]
22. The process according to any of claims 1 to 21, wherein the enzyme is one or more enzymes selected independently from the group consisting of cellulase, hemicellulase, xylanase, glucanase, betaglucanase, pectinase, amylase, alpha-amylase, phospholipase, arabanase, galacto-, beta-mannanase, protease and phytase; wherein the amount of enzyme is 0.5% to 10% by weight of the plant material; and the pH of the mixture is 3-10.
[23]
23. The process of claim 22, wherein the enzyme is a mixture of cellulase, beta-glucanase, pectinase, beta-mannanase, alpha-amylase and protease; wherein the amount of enzyme is 3% by weight of the plant material; and the pH of the mixture is adjusted to pH 5.6 with citric acid monohydrate.
[24]
24. The method of any one of claims 1 to 23, wherein the enzyme concentration and the pH level of the mixture produce optimal enzyme activity.
[25]
A process according to any of claims 1 to 24, wherein the lipid is one or more green and / or food solvents selected independently from the group consisting of olive oil, coconut oil, sesame oil, vegetable oil, milk, butter, liposomes, ethyl acetate, glycerin, d-limonene, butylene glycol, propylene glycol, polyethylene glycol, liposomes, lecithin, ethylhexyl palmitate and
hemp seed or a your mixture.26. Process in according to any of claims 1 to 25, in that the proportion by weight in lipid for plant material in dry matter is situated at
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5/10 range of 0.01: 1 to 4: 1, preferably 2: 3 and the weight ratio of water to plant material to dry matter is in the range of 0.01: 1 to 10: 1, preferably 2: 1.
[26]
27. The method of any one of claims 1 to 26, wherein the weight ratio of lipid to plant material in the dry matter is in the range of 0.1: 1 to 4: 1 and the weight ratio of water for plant material in the dry matter it is in the range of 1: 1 to 5: 1.
[27]
The process according to any of claims 1 to 27, wherein the weight ratio of lipid to plant material in the dry matter is in the range of 0.5: 1 to 1.5: 1 and the weight ratio of water for plant material in the dry matter in the range of 1: 1 to 3: 1.
[28]
29. The method of any one of claims 1 to 28, wherein the mixture is treated with ultrasound or microwave before adding the enzymes.
[29]
The process according to any one of claims 1 to 29, wherein the mixture is treated with ultrasound or microwave after the addition of the enzymes.
[30]
The process according to any one of claims 1 to 30, wherein the lipids, water and enzymes are added in any different order combinations.
32. Process in wake up with any an of claims 1 to 31, in that the steps a. and B. are inverted.33. Process in wake up with any an of
claims 1 to 32, wherein the fat-soluble extract is recirculated any number of times to reach a content
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6/10 higher phyto-cannabinoid and / or terpene / terpenoid.
34. Process in wake up with any an of claims 1 to 33, in that at minus 70% of one mode preferred, more than 80%, in a way more preferred, more of 90% of phyto-cannabinoids s are extracted in extract fat-soluble.35. Process in wake up with any an of
claims 1 to 34, wherein the fat-soluble extract has a total phyto-cannabinoid content of at least 2 weight percent.
[31]
36. The process of any one of claims 1 to 35, wherein the fat-soluble extract has a total phyto-cannabinoid content of at least 3 weight percent.
[32]
37. The process of any one of claims 1 to 36, wherein the fat-soluble extract has a total phyto-cannabinoid content of at least 5 weight percent.
[33]
38. The process according to any one of claims 1 to 37, wherein the ratio between the two main cannabinoids in the fat-soluble extract differs by less than 10%, preferably less than 5%, the ratio between the two main cannabinoids in the plant material .
[34]
39. The process according to any one of claims 1 to 38, wherein less than 10%, preferably less than 5%, more preferably less than 2%, of cannabinoids are decarboxylated during the process.
[35]
40. Process according to any one of claims 1 to 39, wherein at least 70% of the terpenes contained in the plant material are extracted in the extract
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7/10 fat soluble.
[36]
41. The process of any one of claims 1 to 40, wherein at least 70% of the diterpenes contained in the plant material are extracted in the fat-soluble extract.
[37]
42. The process of any one of claims 1 to 41, wherein at least 70% of the sesquiterpenes contained in the plant material are extracted in the fat-soluble extract.
[38]
43. The process of any one of claims 1 to 42, wherein at least 70% of the monoterpenes contained in the plant material are extracted in the fat-soluble extract.
[39]
44. Fat-soluble extract containing phytocannabinoids and / or terpenoids obtainable from the process according to any one of claims 1 to 43.
[40]
45. Fat-soluble extract according to claim 44, wherein the extract has a total cannabinoid content of at least 5 weight percent.
[41]
46. Fat-soluble extract according to either of Claims 44 or 45, with a total cannabinoid content of at least 90%, preferably at least 95% of the initial content after four weeks of browning at 25 ° C.
[42]
47. Fat soluble extract according to any one of claims 44 to 46, wherein the variation in the CBN: THCtot ratio after four weeks in the dark at 25 ° C is less than 5% of the initial proportion.
[43]
48. Fat-soluble extract according to any
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8/10 one of claims 44 to 47, wherein the CBN content is less than 0.1% after four weeks in the dark at 25 ° C.
[44]
49. Fat-soluble extract according to any of claims 44 to 48, wherein the ratio between the two main cannabinoids in the fat-soluble extract differs by less than 10%, preferably less than 5%, the ratio between the two main cannabinoids in the material vegetable.
[45]
50. Fat soluble extract according to any one of claims 44 to 49, wherein the decarboxylated forms of cannabinoids represent less than 10%, preferably less than 5%, more preferably less than 2% of the total cannabinoid content.
[46]
51. Fat-soluble extract according to any one of claims 44 to 50, wherein the terpenoid content is at least 75% of the terpene content of the initial plant material by weight.
[47]
52. Lipid-soluble extract according to any one of claims 44 to 51, wherein the variation in the monoterpene / diterpene ratio after four weeks in the dark at 25 ° C is less than 5% and the monoterpene content is 80% of the initial content.
[48]
53. Fat-soluble extract according to any one of claims 44 to 52, wherein the monoterpene content is at least 30% of the total terpene content.
[49]
54. Fat-soluble extract according to any one of claims 44 to 53, having a cannabinoid content of:
• DELTA-9-TETRAIDROCANABINOL (THC-Acid): in a range of 1,000 to 6,000 mg / kg • DELTA-9-TETRAIDROCANABINOL (THC-NEUTRAL): in a
Petition 870190065723, of 12/07/2019, p. 67/79
9/10 range from 100 to 500 mg / kg • DELTA-9-TETRAIDROCANABINOL (THC-TOTAL EXPRESSED AS NEUTRAL THC): in a range of 1,000 to 7,000 mg / kg • CANABIDIOL (CBD): in a range of 1,000 to 5,000 mg / kg • CANABIDIOL ACID (CBD-A): in a range of 20,000 to 80,000 mg / kg.
[50]
55. Use of the fat-soluble extract according to any one of claims 44 to 54 for the preparation of pharmaceutical or nutraceutical products, cosmetics, food or feed products, antimicrobials, antibacterials, insecticides or biopesticides.
[51]
56. The solid phase resulting from the process according to any of claims 1 to 43, wherein the phyto-cannabinoid content of the plant material is reduced by at least 75 weight percent.
[52]
57. Solid phase according to claim 56, wherein the phyto-cannabinoid content of the plant material is reduced by at least 80 weight percent.
[53]
58. Solid phase according to either of Claims 56 or 57, wherein the phyto-cannabinoid content of plant material is reduced by at least 90 percent by weight.
[54]
59. Use of the solid phase according to any one of claims 56 to 58, for the formulation of food and feed products.
[55]
60. Use of the fat-soluble extract according to any of claims 44 to 54 for the preparation of a cream or gel containing phyto-cannabinoids in which cannabinoids are present in more than 90% of their content
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Initial 10/10 after 10 months.
[56]
61. Use of fat-soluble extract according to any of claims 44 to 54 for the preparation
of a gum or candy containing at least 0.5% phytosterols
cannabinoids, in which phyto-cannabinoids are present in more than 90% of their initial content after 10 months.
[57]
62. Use of the fat-soluble extract according to any of claims 44 to 54 for the preparation of a liposome-based material with at least 0.5% cannabinoids which has been described as having an increased cannabinoid stability of at least 90% of the initial content after 10 weeks.
leave 63. Use of the aqueous phase that can be obtained from the process according to any of the
claims 1 to 43, for the preparation of pharmaceutical or nutraceutical products, cosmetics, antimicrobials, antibacterials, insecticides or biopesticides.

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引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

US6403126B1|1999-05-26|2002-06-11|Websar Innovations Inc.|Cannabinoid extraction method|

---

US9186386B2|2014-04-17|2015-11-17|Gary J. Speier|Pharmaceutical composition and method of manufacturing|

---

US9895404B1|2015-10-09|2018-02-20|Paul T. Baskis|Cannabidiol extraction plant and processes|

---

US9655936B2|2015-10-19|2017-05-23|Aari Ruben|Extraction of cannabidiol|

---

GB2548873B|2016-03-31|2020-12-02|Gw Res Ltd|Use of Cannabidiol in the Treatment of SturgeWeber Syndrome|US10239808B1|2016-12-07|2019-03-26|Canopy Holdings, LLC|Cannabis extracts|

---

EP3745884A1|2018-01-31|2020-12-09|Canopy Holdings, Llc|Hemp powder|

---

US10946307B2|2018-07-12|2021-03-16|Bright Green Corporation|Extraction of cannabinoids, curcuminoids and ginsenosides|

---

US10946308B2|2018-07-12|2021-03-16|Bright Green Corporation|Enzymatic method for extraction and purification of phytocannabinoids|

---

EP3833368A1|2018-08-08|2021-06-16|Neptune Wellness Solutions Inc.|Cold extraction method for cannabinoids and terpenes from cannabis by polyunsaturated lipid-based solvents|

---

WO2020028992A1|2018-08-08|2020-02-13|Neptune Wellness Solutions Inc.|Cold extraction method for cannabinoids and terpenes from cannabis by organic solvents|

---

US11040932B2|2018-10-10|2021-06-22|Treehouse Biotech, Inc.|Synthesis of cannabigerol|

---

WO2020086291A1|2018-10-22|2020-04-30|Jones Lisa Marie|Topical compositions incorporating cannabis|

---

EP3894572A1|2018-12-10|2021-10-20|Loxley Systems, LLC|Systems, methods, and equipment for chemical extraction|

---

WO2021037343A1|2019-08-27|2021-03-04|Herbolea Biotech S.R.L.|Cannabinoid concentrate and isolate, method of obtaining the same and use thereof|

---

US10765965B1|2019-12-10|2020-09-08|Loxley Systems, Llc|Systems, methods, and equipment for chemical extraction|

---

WO2022043422A1|2020-08-26|2022-03-03|K.D. Pharma Bexbach Gmbh|Extraction method and mixture of substances|

法律状态:
2021-10-13| B350| Update of information on the portal [chapter 15.35 patent gazette]|

优先权:

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申请号 | 申请日 | 专利标题

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US201762446429P| true| 2017-01-14|2017-01-14|

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US62/446,429|2017-01-14|

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US201762524239P| true| 2017-06-23|2017-06-23|

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US62/524,239|2017-06-23|

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US201762546372P| true| 2017-08-16|2017-08-16|

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US62/546,372|2017-08-16|

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PCT/EP2018/050860|WO2018130682A1|2017-01-14|2018-01-15|Enzyme-assisted lipid-based extraction and stabilization of phyto-cannabinoids and terpens and products obtained thereof|

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