CN114685416B - Separation method of cannabinoid - Google Patents

Separation method of cannabinoid Download PDF

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Publication number
CN114685416B
CN114685416B CN202011613795.5A CN202011613795A CN114685416B CN 114685416 B CN114685416 B CN 114685416B CN 202011613795 A CN202011613795 A CN 202011613795A CN 114685416 B CN114685416 B CN 114685416B
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cannabinol
separating
raw material
solid phase
phase extraction
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CN114685416A (en
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柳旭
赵丽梅
高伟博
常坦然
李如彥
李庆中
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Yunnan Hanmeng Pharmaceutical Co ltd
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Yunnan Hanmeng Pharmaceutical Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/78—Ring systems having three or more relevant rings
    • C07D311/80—Dibenzopyrans; Hydrogenated dibenzopyrans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/94—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems condensed with rings other than six-membered or with ring systems containing such rings

Abstract

The application provides a separation method of cannabinoid. The cannabinoid separation method specifically comprises the following steps: (1) Purifying and concentrating the raw material sample through solid phase extraction to obtain a section I containing tetrahydrocannabinol and cannabinol and a section II containing cannabichromene and cannabinol; (2) Separating and purifying the I section containing tetrahydrocannabinol and cannabinol and the II section containing cannabinol and cannabinol by column chromatography to obtain primary tetrahydrocannabinol, primary cannabinol and primary cannabinol. The application adopts a solid phase extraction purification mode to effectively remove wax, grease and pigment, the sectional purification effect is superior to other technical means, and the application combines the conventional column chromatography separation purification preparation to realize the purposes of recovering and separating cannabidiol, cannabinol, cannabidene and cannabinol with high purity and high yield, and reduces the destruction cost of the tail material containing tetrahydrocannabinol.

Description

Separation method of cannabinoid
Technical Field
The application relates to the technical field of extraction and separation of natural phytochemicals, in particular to a separation method of magnetic cannabinoids.
Background
Industrial Cannabis (academic name: cannabis sativa L.) is a plant of Cannabiaceae and Cannabis, and Tetrahydrocannabinol (THC) content in flowers and leaves in growth period is less than three thousandths, so that large-scale planting and industrialized development and utilization can be legally performed. The flowers and leaves of industrial cannabis contain cannabinoids and non-cannabinoids and are widely used in the medical field. Wherein the cannabinoid family is mainly classified into Tetrahydrocannabinol (THC), cannabinol (CBN), cannabidiol (CBD), cannabidiol (CBG), hypocreosol (CBDV), cannabidene (CBC), hypocreosol (THCV), tetrahydrocannabinol, and the like; the main classes of non-cannabinoids are: ephedrones, other terpenoids, beta sitosterols; these extracts are widely used in the medical industry.
With the continued development of Cannabidiol (CBD), the medicinal value of other related cannabinoids is also becoming increasingly recognized. For example, patent US8470874B2 indicates that natural or synthetic CBCs and derivatives thereof prepared therefrom are particularly effective in the treatment of mood disorders such as depression and show a strong effect in anti-inflammatory and antibacterial aspects. Cannabinol (CBL) is more reported as an active ingredient, and there is no pharmacological efficacy data of a single component. Patent US20030158191A1 indicates the use of Cannabinol (CBN) and its derivatives in the treatment of diseases of the immune system such as hiv and oncological diseases.
However, the fraction of hemp extract tails often contains large amounts of oil waxes and pigments and is complex in composition. Currently, the most advanced treatment is molecular distillation, from which cannabinoids can be separated as the light phase, but provided that the distillation must be carried out with preferential removal of the wax and that it only gives a mixture of one boiling range. At present, no industrial preparation method for materials with the characteristics exists.
CN112010738A discloses a chromatographic separation method of cannabinoid compounds, comprising chromatographic separation of cannabis extract with aqueous ethanol solution, comprising the steps of: (1) washing impurities, namely washing impurities by adopting an ethanol water solution; (2) Gradient elution, namely, adopting an ethanol water solution as a gradient elution solvent, and respectively collecting the eluent of 1-2BV, the eluent of 2.5-4BV and the eluent of 4-6 BV; (3) Concentrating, namely concentrating the eluent collected in the step (2) respectively. The method can directly separate high-purity cannabidiol, cannabidiol and tetrahydrocannabinol; the method is carried out in a reverse separation mode in the whole process for improving the purity, but certainly the reverse separation is not applicable to the factory tailings containing a large amount of wax, pigment and grease, and the reverse separation mode in the whole process also brings great difficulty to post-treatment concentration, so that the method is not applicable to industrial production.
CN111315459a discloses a process for purifying and separating cannabinoids from dried cannabis and cannabis leaves, which uses simulated moving bed chromatography for continuous purification of cannabinoids extracted from dried cannabis and cannabis leaves, which purifies cannabinoids, in particular cannabidiol and tetrahydrocannabinol, by filtration, decolorization, activation or decarboxylation, dewaxing, polishing and crystallization purification steps as well as continuous simulated moving bed process recovery steps, which is complicated for waxy, pigment and grease removal processes and fails to recover pure products of cannabinol, cannabichromene and cannabinol.
Therefore, development of a process suitable for industrial treatment of process plant tailings and efficient recovery and separation of various cannabinoids is a focus of research in the art.
Disclosure of Invention
Aiming at the defects existing in the prior art, the application aims to provide a separation method of cannabinoid. The separation method provided by the application realizes the purposes of recovering and separating cannabidiol, cannabinol, cannabidene and cannabinol with high purity and high yield, and reduces the destruction components of the tail material containing tetrahydrocannabinol.
To achieve the purpose, the application adopts the following technical scheme:
in a first aspect, the present application provides a method for separating cannabinoids, comprising the steps of:
(1) Purifying and concentrating the raw material sample through solid phase extraction to obtain a section I containing tetrahydrocannabinol and cannabinol and a section II containing cannabichromene and cannabinol;
(2) Separating and purifying the I section containing tetrahydrocannabinol and cannabinol and the II section containing cannabinol and cannabinol by column chromatography to obtain primary tetrahydrocannabinol, primary cannabinol and primary cannabinol.
In the application, the wax, grease and pigment are effectively removed by adopting a solid phase extraction purification mode, the sectional purification effect is superior to that of other technical means (such as molecular distillation), and then the method is combined with the conventional column chromatography separation purification preparation, so that the purposes of recovering and separating cannabidiol, tetrahydrocannabinol, cannabinol, cannabinene and cannabinol with high purity and high yield are realized, the destruction components of the tail material containing tetrahydrocannabinol are reduced, and the method can realize production amplification.
Preferably, in the step (1), the mass content of tetrahydrocannabinol in the raw material sample is 0.3% or less, for example, 0.3%, 0.25%, 0.2%, 0.15%, 0.1%, 0.05%, etc.
Preferably, in the step (1), the mass content of cannabidiol in the raw material sample is 2-20%, the mass content of cannabidiol is 1-10%, the mass content of cannabidene is 0.5-5%, and the mass content of cannabinol is 2-20%.
The cannabidiol content is 2-20% by mass, based on 100% by mass of the raw material sample, and may be, for example, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20% or the like.
The mass content of the cannabinol is 1-10% based on 100% of the mass of the raw material sample, and may be, for example, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or the like.
The mass content of the cannabidene is 0.5-5% based on 100% of the mass of the raw material sample, and may be, for example, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5% or the like.
The mass content of the cannabinol is 2-20% based on 100% of the mass of the raw material sample, and may be, for example, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20% or the like.
Preferably, in step (1), the feedstock sample is cannabidiol extraction plant tailings or industrial cannabis extract feedstock.
Wherein, the cannabidiol extraction plant tailings refer to the residual parts collectively called extraction plant tailings after CBD extraction is complete; industrial cannabis extract starting material refers to an alcoholic extract of CBD or other organic solvent extract. I.e. the raw material sample can be industrial hemp with THC content below 0.3%, and the residual tailing is after CBD is extracted by the pharmaceutical extraction factory of China Union; or mature hemp flowers and leaves with 7-9 months of flower and leaf raw materials, and is primarily dried by farmers, and is also suitable for preparing industrial hemp alcohol extract with THC content below 0.3%.
Preferably, in step (1), the feedstock sample is selected from any one or a combination of at least two of industrial hemp, intermediate hemp or medicinal hemp;
preferably, in the step (1), the extracted part of the industrial hemp extract raw material is any one or a combination of at least two of hemp flowers, hemp leaves, hemp roots, hemp stalk cores and hemp seed meal.
Preferably, in the step (1), the solid phase extraction is performed under a low temperature condition of-10 to 10 ℃, for example, -10 ℃, -5 ℃, 0 ℃, 5 ℃, 10 ℃ and the like.
The low temperature condition is that cold circulation liquid at the temperature of minus 10 ℃ to 10 ℃ is introduced into the chromatographic column jacket, so that wax, pigment and most of grease are deposited at the top end of column packing, and the subsequent treatment is easy.
Preferably, in the step (1), the filler used for the solid phase extraction purification comprises any one or a combination of at least two of activated clay, diatomaceous earth, calcium phosphate, calcium carbonate, polyamide powder and silica gel.
The filler adsorbent used in the application can be recycled after being dried (such as clay and calcium phosphate can be used for picking out the top wax and other polluted parts, and can be used repeatedly after being treated for 1-5 hours at 300-500 ℃), and the theoretical time can reach more than 100 times, and basically no production waste is generated.
Preferably, the particle size of the polyamide powder is 90 to 120 mesh, for example, 90 mesh, 95 mesh, 100 mesh, 105 mesh, 110 mesh, 115 mesh, 120 mesh, etc.
Preferably, the silica gel has a particle size of 100 to 500 mesh, for example, 100 mesh, 200 mesh, 300 mesh, 400 mesh, 500 mesh, etc.
Preferably, the particle size of the above-mentioned other fillers, except for the polyamide powder and the silica gel, is 30 to 300 mesh, for example, 30 mesh, 40 mesh, 50 mesh, 60 mesh, 80 mesh, 100 mesh, 150 mesh, 200 mesh, 250 mesh, 300 mesh, etc. each independently.
Preferably, in step (1), the solid phase extraction purification is performed using a solid phase extraction column.
Preferably, in step (1), the diameter-to-height ratio of the solid phase extraction column is 1 (1-3), and may be, for example, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, etc.
In the application, the diameter-to-height ratio of the solid phase extraction column is limited to be 1 (1-3), the requirement of preliminary separation can be met, if the extraction column is smaller in the shorter diameter height, the separation degree is reduced, and the crossed overlapping limit of each separation section is not clear; if the extraction column is smaller in diameter and larger in height, the separation effect is improved, but the separation is slow, the column blockage risk is more likely to occur, and the auxiliary flushing with larger positive air pressure is required, so that the waste of the filler adsorbent is caused.
Preferably, in the step (1), the mass ratio of the total amount of the filler to the raw material sample in the solid phase extraction column is 1 (0.25-5), for example, may be 1:0.25, 1:0.3, 1:0.4, 1:0.5, 1:1, 1:1.2, 1:1.4, 1:1.6, 1:1.8, 1:2, 1:2.5, 1:3, 1:4, 1:5, and the like, preferably 1 (0.5-2).
Preferably, in step (1), the solid phase extraction is performed by dry loading or wet loading.
Preferably, the dry loading method specifically comprises the following steps: mixing part of the filler with the raw material sample to form a solid dispersion system, and transferring the solid dispersion system into a solid phase extraction column filled with the filler; the mass ratio of the partial filler to the raw material sample is 1 (0.25-2), for example, 1:0.25, 1:0.3, 1:0.4, 1:0.5, 1:1, 1:1.2, 1:1.4, 1:1.6, 1:1.8, 1:2, and the like, and preferably 1 (0.5-1).
Preferably, the wet loading specifically includes: mixing an organic solvent with a raw material sample to form a liquid dispersion system, and transferring the liquid dispersion system into a solid phase extraction column filled with a filler; the volume/mass ratio of the organic solvent to the raw material sample is 1mL (0.25-5) g, for example, 1mL (0.25-2) g, 1mL (0.5-5) g, 1mL (1 mL) 1g, 1mL (1.5-5) g, 1mL (2 g), 1mL (3 g), 1mL (4 g), 1mL (5-2) g, etc.
Preferably, the organic solvent for wet loading is an eluting solvent, preferably any one or a combination of at least two of hexane, heptane or an aqueous ethanol solution with a concentration of 40-65v% (for example, 40v%, 45v%, 50v%, 55v%, 60v%, 65v%, etc.).
Preferably, the wet sample application is assisted by heating during dissolution or slurry mixing at 50-75deg.C, such as 50deg.C, 55deg.C, 60deg.C, 65deg.C, 70deg.C, 75deg.C, etc.
Preferably, in step (1), the eluting solvent used for the solid phase extraction purification comprises any one or a combination of at least two of alkanes, halogenated alkanes, esters, alcohols, ketones, organic acids or water.
Preferably, the alkane is selected from any one or a combination of at least two of petroleum ether, diethyl ether, n-hexane or n-heptane.
Preferably, the halogenated alkane is selected from dichloromethane and/or chloroform.
Preferably, the ester is selected from ethyl acetate and/or butyl acetate.
Preferably, the alcohol is selected from any one or a combination of at least two of methanol, ethanol or isopropanol.
Preferably, the ketone is selected from acetone and/or butanone.
Preferably, the organic acid is selected from any one or a combination of at least two of formic acid, acetic acid or trifluoroacetic acid.
In the present application, the eluting solvent used may be a binary solvent system such as a mixture of n-hexane and ethyl acetate, a mixture of ethyl acetate and acetone, or 45 to 100% by volume (for example, 45% by volume, 50% by volume, 60% by volume, 70% by volume, 80% by volume, 90% by volume, 100% by volume, etc.) of an aqueous ethanol solution, 85% by volume or more (for example, 85% by volume, 90% by volume, 95% by volume, etc.) of an aqueous isopropanol solution; the eluting solvent used may also be a ternary solvent system, such as mixtures of n-hexane, ethyl acetate and formic acid, ethanol, formic acid and water, and the like.
Preferably, the elution is carried out under medium-low pressure of 0.05-15Mpa, for example, 0.05Mpa, 0.06Mpa, 0.08Mpa, 0.1Mpa, 0.2Mpa, 0.4Mpa, 0.6Mpa, 0.8Mpa, 1Mpa, 2Mpa, 4Mpa, 6Mpa, 8Mpa, 10Mpa, 12Mpa, 14Mpa, 15Mpa, etc.
Preferably, in the step (1), in the solid phase extraction and purification process, the solvent I is used for eluting to obtain the residual cannabidiol in the raw material sample, the solvent II is used for eluting to obtain the I section containing tetrahydrocannabinol and cannabinol, and the solvent III is used for eluting to obtain the II section containing cannabichromene and cannabinol.
Preferably, in step (1), the solvent one is a mixed solution of alcohol or ketone, organic acid and water. That is, the solvent I may be a mixed solution of alcohol, organic acid and water or a mixed solution of ketone, organic acid and water.
Preferably, in step (1), the first solvent comprises, in percentage by volume, based on 100% of the total volume of the first solvent: 45-90% (e.g., 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, etc.), 0.05-5% (e.g., 0.05%, 0.1%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, 5%, etc.), and the balance water.
Preferably, the organic acid is formic acid, and the volume content of the formic acid is 0.5-3%, for example, may be 0.5%, 1%, 2%, 3%, etc. The organic acid may promote more definite elution fractions and more concentrated target components.
Preferably, in the step (1), the solvent two is a mixed solution of alcohol and ester, a mixed solution of ketone and ester, an aqueous solution of alcohol or an aqueous solution of ketone.
Preferably, the volume ratio of the alcohol to the ester is (30-60): 1, which may be, for example, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, etc., and the volume ratio of the ketone to the ester is (30-60): 1, which may be, for example, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, etc.
Preferably, the volume of the alcohol in the aqueous solution of the alcohol is 90-100%, for example, 90%, 92%, 94%, 96%, 98%, 100% or the like.
Preferably, the volume of ketone in the aqueous solution of ketone is 70-90%, for example, 70%, 75%, 80%, 85%, 90%, etc.
Preferably, in step (1), the solvent III is any one or a combination of at least two of alkane, halogenated alkane or ester.
Preferably, in the step (1), the temperature of the concentration is 50-65 ℃, for example, 50 ℃, 52 ℃, 55 ℃, 58 ℃, 60 ℃, 63 ℃, 65 ℃ and the like, and the pressure of the concentration is-0.10 to-0.065 MPa, for example, -0.10MPa, -0.09MPa, -0.08MPa, -0.07MPa, -0.065MPa and the like.
Preferably, in the step (1), the concentration is performed to a dry extract with a relative density of 1.05-1.15, for example, 1.05, 1.06, 1.07, 1.08, 1.09, 1.10, 1.12, 1.14, 1.15, etc.
The term "dry extract" refers to a state without distillate in 5 minutes of vacuum concentration.
Preferably, in step (2), the sections I and II are separated and purified independently by forward column chromatography or reverse column chromatography.
Preferably, the reverse column chromatography adopts any one of reverse ODS silica gel (such as C4, C8, C18, amino, glycosyl, phenyl series, etc.), polyamide powder or MCI resin powder containing special bonding.
Preferably, the reverse-phase column chromatography eluent is selected from any one or a combination of at least two of alcohol, ketone, ester or water.
Preferably, step (2) further comprises step (3): after cracking and destroying the primary tetrahydrocannabinol product, neutral adjustment or inactivation is carried out and then the primary tetrahydrocannabinol product is discarded; concentrating and drying primary cannabinol, primary cannabinol and primary cannabinol respectively.
Preferably, in the step (3), the destruction is performed by a chemical reaction method of oxidation, reduction, enzymatic method, acid-base cleavage or thermal cleavage.
Preferably, in the step (3), the temperature of the concentration is 50-65deg.C, for example, 50 ℃, 52 ℃, 55 ℃, 58 ℃, 60 ℃, 63 ℃, 65 ℃, etc., and the pressure of the concentration is-0.10-0.065 MPa, for example, -0.10MPa, -0.09MPa, -0.08MPa, -0.07MPa, -0.065MPa, etc.
Preferably, in step (3), the drying includes any one or a combination of at least two of vacuum drying, freeze drying, near infrared drying, air drying or microwave drying.
Preferably, in the step (3), the drying temperature is 30 to 65 ℃, for example, 30 ℃, 35 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃ and the like can be used.
Compared with the prior art, the application has the following beneficial effects:
(1) The application provides a separation and purification process of high-purity CBC, CBN and CBL, which has higher transfer rate and purity of more than 95 percent, and simultaneously, the THC part to be destroyed is purified and enriched again, and the destruction cost of Tailing (THC) of more than 80 percent is reduced.
(2) The application adopts the solid phase extraction with controllable temperature, can effectively remove wax, grease and pigment, has the sectional purification effect superior to other technical means (such as molecular distillation), can realize production amplification by combining with the conventional chromatographic preparation, and has excellent industrial prospect.
(3) The adsorbent used in the application can be recycled after being dried (such as clay and calcium phosphate can be used for drawing out the top wax and other polluted parts, and can be used repeatedly after being treated for 1-5 hours at 300-500 ℃), and the theoretical time can reach more than 100 times, and basically no production waste is generated.
Drawings
FIG. 1 is a high performance liquid chromatogram of the CBD extraction plant tailings provided in example 1.
Fig. 2 is a high performance liquid chromatogram of section I containing THC and CBN provided in example 1.
FIG. 3 is a high performance liquid chromatogram of section II containing CBC and CBL provided in example 1.
Fig. 4 is a high performance liquid chromatogram of the finished CBC product provided in example 1.
Fig. 5 is a high performance liquid chromatogram of the finished CBL product provided in example 1.
Fig. 6 is a high performance liquid chromatogram of the finished CBN product provided in example 1.
Detailed Description
The technical scheme of the application is further described below by the specific embodiments with reference to the accompanying drawings. It should be apparent to those skilled in the art that the detailed description is merely provided to aid in understanding the application and should not be taken as limiting the application in any way.
Example 1
The embodiment provides a method for separating cannabinoid, which specifically comprises the following steps:
(1) Taking 100g of CBD extraction plant tailing (wherein the CBD content in the CBD extraction plant tailing is 2.75wt%, the THC content is 8.85wt%, the CBN content is 10.57wt%, the CBC content is 2.92wt%, and the CBL content is 21.43wt%, as shown in figure 1), adding 50mL of n-hexane, pulping under the assistance of 60 ℃, adding 100g of activated clay while the mixture is hot, uniformly mixing, and cooling to room temperature to form a solid dispersion system; the above solid dispersion was transferred to a solid phase extraction column (diameter to height ratio 4:5.5, 240 mL) containing 75g of activated clay and 25g of diatomaceous earth, and the column was washed with a 65v% aqueous ethanol solution at a column temperature of 10℃for 300mL to recover 4.2g of CBD fraction (CBD content 65.53%) for use; then, flushing 500mL of the column with 80v% ethanol water solution to obtain an I section containing THC and CBN; then compressed air is introduced to press the mixture to be dry, and 350mL of the column is punched by normal hexane to obtain a section II containing CBC and CBL; finally, concentrating at 60 ℃ and minus 0.08mpa respectively to obtain 40.36g of target section I eluate (shown in figure 2) and 26.88g of section II eluate (shown in figure 3).
(2) Subjecting the eluate of the section I to 200-300 mesh silica gel column (diameter-height ratio is 3:45, 320 mL), and sequentially eluting with mixed solution (450 mL) of n-hexane, ethyl acetate and formic acid at volume ratio of 99.45:0.5:0.05 to obtain THC primary product; obtaining a CBN primary product solution by mixing n-hexane, ethyl acetate and formic acid (550 mL) with the volume ratio of 98.45:1.5:0.05;
loading the eluate of section II onto silica gel column (diameter-to-height ratio of 2:35, 105 mL), and sequentially eluting with n-hexane (200 mL) to obtain CBL primary product solution; the CBC primary product solution is obtained by eluting with a mixed solution (150 mL) of normal hexane and ethyl acetate with the volume ratio of 98.5:1.5.
(3) Combining THC-containing components, adding 11.20g in total, destroying by nitric acid pyrolysis, adjusting pH to 7.0, and discarding; concentrating the other three primary products into paste at 65 ℃ and minus 0.09MPa, and vacuum drying the three primary products at 50 ℃ and minus 0.10MPa for 48 hours to obtain 9.66g of CBN, 20.01g of CBL and 2.65g of CBC finished product. Samples were taken and assayed by HPLC for a CBC purity of 98.25% (as shown in FIG. 4), a CBL purity of 99.26% (as shown in FIG. 5) and a CBN purity of 98.87% (as shown in FIG. 6).
Example 2
The embodiment provides a method for separating cannabinoid, which specifically comprises the following steps:
(1) 1000g of an industrial hemp leaf alcohol extract (the industrial hemp leaf alcohol extract contains 14.55wt% of CBD, 0.25wt% of THC, 7.55wt% of CBN, 1.51wt% of CBC and 1.41wt% of CBL) is added into 1000mL of a 60v% ethanol water solution, and the mixture is homogenized at 45 ℃ to form a liquid dispersion system; transferring the above liquid dispersion to a solid phase extraction column (diameter-height ratio 40:55, 6L) filled with 2000g polyamide powder (90-120 mesh), flushing column 12L with 45v% ethanol water solution at 0deg.C, and recovering 278g (CBD content 52.18%) rich in CBD and CBDV; flushing the column by using a 90v% ethanol water solution for 10L to obtain an I section containing THC and CBN; finally, introducing compressed air, drying, and flushing a column 9L with ethyl acetate to obtain a section II containing CBC and CBL; concentrating at 50 ℃ and-0.09 MPa respectively to obtain 121.76g of target section I eluate and 75.83g of target section II eluate.
(2) The eluate of the section I is put on an MCI column (75-120 μm, diameter-to-height ratio is 6:45, 1.28L), and 2.8L is eluted with 85v% isopropyl alcohol water solution at equal degree, so as to obtain THC primary product solution and CBN primary product solution in sequence; the above-mentioned eluate of section II is put on MCI column (75-120 μm, diameter-height ratio is 6:45, 1.28L), and 1.8L is isocratically eluted with 85v% isopropyl alcohol aqueous solution to obtain CBC primary product solution, and 1.6L is flushed with isopropyl alcohol to obtain CBL primary product solution.
(3) Combining all THC-containing components to obtain 57.52g, thermally cracking at 350 ℃, destroying, and discarding; concentrating the other three primary product solutions at 65 deg.C and-0.09 MPa to obtain paste, vacuum drying at 50 deg.C and-0.10 MPa for 48h, and drying to obtain final product of CBN 60.72g,CBL 13.18g and CBC 12.75 g. Samples were taken, and the purity of CBN was 99.05%, CBL 98.97% and CBC 98.86% as determined by HPLC.
Example 3
The embodiment provides a method for separating cannabinoid, which specifically comprises the following steps:
(1) 1000g of CBD extraction plant tailing (wherein the CBD content in the CBD extraction plant tailing is 2.82wt%, the THC content is 9.75wt%, the CBN content is 11.57wt%, the CBC content is 2.96wt%, and the CBL content is 21.43 wt%) is added with 500mL of n-heptane, and the mixture is homogenized under the assistance of 50 ℃ to form a liquid dispersion system; the above liquid dispersion was transferred to a solid phase extraction cartridge (aspect ratio 10:30, 1.2L) containing 1.62kg of calcium phosphate and 0.18kg of diatomaceous earth (120-200 mesh); after the temperature of the column is reduced to-10 ℃, 1.8L of ethanol is used for flushing the column, 48.06g (57.98 percent of CBD content) of components rich in CBD and the like are recovered for standby; introducing compressed air to press dry the chromatographic column, and flushing the column with a mixed solution (2.5L) of n-heptane, ethyl acetate and formic acid in a volume ratio of 99:0.5:0.5 to obtain a section I containing CBC and CBL; flushing a column by using a mixed solution (2L) of n-heptane and ethyl acetate with the volume ratio of 90:10 to obtain a section II containing THC and CBN; concentrating at 50 ℃ and-0.09 MPa respectively to obtain 376.58g of target section I eluate and 240.27g of target section II eluate.
(2) The eluate of the section I is put on an MCI column (75-120 μm, length-diameter ratio is 6:45, 1.28L), 2.4L is eluted with 90v% ethanol water solution isocratically to obtain a CBC primary product solution, and 1.8L is washed with ethyl acetate to obtain a CBL primary product solution. Loading the eluate of the section II onto YMC 4 silica gel chromatographic column (60-210 μm, length-diameter ratio of 6:45, 1.2L), and isocratically eluting with a mixed solution of n-heptane, ethyl acetate and formic acid with volume ratio of 98:1.5:0.5 for 3.2L to obtain THC primary product solution and CBN primary product solution;
(3) Combining all THC-containing components, adding 146.25g in total, cracking and destroying with acidic potassium permanganate at 70 ℃, and discarding after neutral adjustment; concentrating the other three primary products at 65 deg.C and-0.09 Mpa to obtain paste, vacuum drying the above three primary products at 50 deg.C and-0.10 MPa for 48h to obtain 112.30gCBN,202.11g CBL and 26.85g CBC. Samples were taken and the purity of CBC was 98.61%, CBL 99.66% and CBN 99.19% as determined by HPLC.
Example 4
This example provides a method for separating cannabinoids, which differs from example 1 in that the aspect ratio of the solid phase extraction cartridge in step (1) is 1:0.8, and the other steps are the same as in example 1.
Example 5
This example provides a method for separating cannabinoids, which differs from example 1 in that the aspect ratio of the solid phase extraction cartridge in step (1) is 1:5, and the other steps are the same as in example 1.
Example 6
This example provides a method for separating cannabinoid, which is different from example 1 in that 100g of CBD extraction plant tailing is taken in step (1), 50mL of normal hexane is added, pulp is stirred under the assistance of 60 ℃, then 25g of activated clay is added while the mixture is hot, and the other steps are the same as example 1.
Example 7
This example provides a method for separating cannabinoid, which is different from example 1 in that 100g of CBD extraction plant tailing is taken in step (1), 50mL of normal hexane is added, pulp is stirred under the assistance of 60 ℃, then 200g of activated clay is added while the mixture is hot, and the other steps are the same as example 1.
Example 8
The present example provides a method for separating cannabinoid, which is different from example 1 in that in step (2), the above-mentioned eluate of section I is subjected to 200-300 mesh silica gel column (diameter-to-height ratio 3:45, 320 mL), and then eluted sequentially with a mixed solution (450 mL) of n-hexane and ethyl acetate in a volume ratio of 99.5:0.5 to obtain THC as a primary product, and the other steps are the same as in example 1.
Example 9
The present example provides a method for separating cannabinoid, which is different from example 1 in that in step (2), a CBN primary product solution is obtained by mixing (550 mL) n-hexane and ethyl acetate in a volume ratio of 98.5:1.5.
Comparative example 1
The comparative example provides a method for separating cannabinoid, which specifically comprises the following steps:
(1) Taking 100g of CBD extraction plant tailing, wherein the content of CBD is 2.75%, the content of THC is 8.85%, the content of CBN is 10.57%, the content of CBC is 2.92% and the content of CBL is 21.43%. Adding 50mL of n-hexane, pulping at 60 ℃ in an auxiliary way, adding 100g of activated clay while the mixture is hot, uniformly mixing, and cooling to room temperature to form a solid dispersion system; the above solid dispersion was transferred to a solid phase extraction column (diameter to height ratio 4:5.5, 240 mL) containing 75g of activated clay and 25g of diatomaceous earth, and the column was washed with a 65v% aqueous ethanol solution at a column temperature of 10℃for 300mL to recover 4.2g of CBD fraction (CBD content 65.53%) for use; compressed air is then introduced and dried by pressing with n-hexane 98: the solvent was recovered by washing with 780mL ethyl acetate, and concentrating under reduced pressure to obtain 68.62g of a mixed eluate containing THC, CBN, CBC and CBL.
(2) Loading the mixed eluate onto 200-300 mesh silica gel column (diameter-height ratio of 3:45, 320 mL) in sequence; eluting with n-hexane (200 mL) to obtain a CBL primary product solution; eluting with a mixed solution (450 mL) of n-hexane, ethyl acetate and formic acid in a volume ratio of 99.45:0.5:0.05 to obtain a THC primary product; eluting with a mixed solution (250 mL) of n-hexane, ethyl acetate and formic acid in a volume ratio of 98.45:1.5:0.05 to obtain a CBC primary product solution; a mixed solution (500 mL) of n-hexane, ethyl acetate and formic acid with the volume ratio of 98.45:1.5:0.05 is used for obtaining a CBN primary product solution.
(3) Combining THC-containing components, adding 20.17g in total, destroying by nitric acid pyrolysis, adjusting pH to 7.0, and discarding; in addition, the other three primary product solutions are concentrated into paste at 65 ℃ and minus 0.09MPa respectively, and the three primary product pastes are dried in vacuum at 50 ℃ and minus 0.10MPa for 48 hours, thus obtaining 13.78g of CBN, 25.79g of CBL and 4.29g of CBC finished product. Samples were taken, and the purity of CBC was 58.97%, the purity of CBL was 79.19% and the purity of CBN was 63.92% as determined by HPLC.
Comparative example 2
The comparative example provides a method for separating cannabinoid, which specifically comprises the following steps:
(1) Taking 100g of CBD extraction plant tailing, wherein the content of CBD is 2.75%, the content of THC is 8.85%, the content of CBN is 10.57%, the content of CBC is 2.92% and the content of CBL is 21.43%. Adding 150mL of n-hexane, pulping under the assistance of 60 ℃, adding a 200-300 mesh silica gel column (with the diameter-height ratio of 3:45, 320 mL) while the mixture is hot, cooling to room temperature (25.5 ℃) after the sample is completely loaded, and eluting with n-hexane (450 mL) in sequence to obtain a CBL primary product solution; eluting with a mixed solution (450 mL) of n-hexane, ethyl acetate and formic acid in a volume ratio of 99.45:0.5:0.05 to obtain a THC primary product; eluting with a mixed solution (320 mL) of n-hexane, ethyl acetate and formic acid in a volume ratio of 98.45:1.5:0.05 to obtain a CBC primary product solution; a mixed solution (500 mL) of n-hexane, ethyl acetate and formic acid with the volume ratio of 98.45:1.5:0.05 is used for obtaining a CBN primary product solution.
(2) Combining THC-containing components, adding 25.33g total, destroying by nitric acid pyrolysis, adjusting pH to 7.0, and discarding; in addition, the other three primary product solutions are concentrated into paste at 65 ℃ and minus 0.09MPa respectively, and then the three primary product pastes are dried in vacuum at 50 ℃ and minus 0.10MPa for 48 hours, thus obtaining 17.66g of CBN, 38.15g of CBL and 8.05g of CBC finished product. Samples were taken and the purity of CBC was 31.04%, CBL 53.85% and CBN 48.79% by HPLC.
Performance testing
The purity test and recovery calculations were performed on the finished products provided in examples 1-9 and comparative examples 1-2, respectively, using HPLC, as follows:
chromatographic conditions and system suitability test: octadecylsilane chemically bonded silica is used as a filler; acetonitrile as mobile phase a and 0.1% formic acid-water as mobile phase B, gradient elution was performed according to the following a (%): B (%) schedule: eluting for 0 to 11.5min at a ratio of 72:28; 11.5min to 24min up to 90:10; reducing the time from 24min to 26min to 72:28; the detection wavelength was 210nm. The theoretical plate number is not lower than 2500 calculated according to the CBN peak;
preparation of a control solution: accurately weighing CBN, CBC, CBL, THC, CBD reference substance, and adding methanol-water (1:1) to obtain mixed reference substance solution containing 0.03mg per L.
Preparation of test solution: taking raw materials (tailing sample/extract sample) or three finished cannabinoid products after drying, precisely weighing about 8-12mg, placing in a 250mL measuring flask, adding 20mL of acetonitrile, carrying out ultrasonic treatment for 10 minutes, adding acetonitrile again to dilute to a scale, shaking uniformly, filtering with a microporous filter membrane (0.45 pm), and taking the subsequent filtrate to obtain the final product.
Assay: respectively precisely sucking 10 μl of each of the reference solution and the sample solution, and injecting into a liquid chromatograph for measurement.
Raw material of cannabinoid XX purity= (sample XX peak area×0.03x250)/(reference sample XX peak area×sample actual weighing value) ×100%
The recovery of certain cannabinoid XX is calculated as follows:
recovery of certain cannabinoid XX = (finished product weight x purity of certain cannabinoid XX)/(weight of starting material x purity of certain cannabinoid XX) ×100%
Wherein, a cannabinoid XX refers to one of CBN, CBC, CBL, THC, CBD.
The specific test results are shown in table 1:
TABLE 1
As shown in the test data of Table 1, the purity of CBC, CBL and CBN obtained by the cannabinoid separation method can reach more than 98%, and the recovery rate can reach more than 89%, so that the temperature-controlled solid phase extraction technology of the application is demonstrated, the segmentation and fine separation of complex components can be realized by combining a specific chromatographic method, and finally, various cannabinoid finished products with the purity of more than 95% can be obtained, the recovery rate is more than 70%, and the process is relatively simple.
The applicant states that the present application describes the isolation method of said cannabinoids by means of the above examples, but the present application is not limited to, i.e. it is not meant that the present application must be carried out in dependence on the above examples. It should be apparent to those skilled in the art that any modification of the present application, equivalent substitution of each raw material sample of the product of the present application, addition of auxiliary components, selection of specific modes, etc., fall within the scope of the present application and the scope of disclosure.

Claims (23)

1. A method for separating cannabinoids, comprising the steps of:
(1) Purifying and concentrating the raw material sample through solid phase extraction to obtain a section I containing tetrahydrocannabinol and cannabinol and a section II containing cannabichromene and cannabinol; the solid phase extraction is carried out under the low temperature condition, wherein the low temperature condition is-10 ℃; the solid phase extraction and purification is carried out by adopting a solid phase extraction column, wherein the diameter-to-height ratio of the solid phase extraction column is 1 (1-3), and the mass ratio of the total amount of filler in the solid phase extraction column to the raw material sample is 1 (0.25-5); in the solid phase extraction and purification process, firstly eluting with a first solvent to obtain residual cannabidiol in a raw material sample, then eluting with a second solvent to obtain a section I containing tetrahydrocannabinol and cannabinol, and finally eluting with a third solvent to obtain a section II containing cannabichromene and cannabinol; the raw material sample is cannabidiol extraction factory tailings or industrial cannabis extract raw material; the first solvent is ethanol, 45v% ethanol water solution or 65v% ethanol water solution; the solvent II is an aqueous solution of 80% by volume and 90-100% by volume of alcohol or a mixed solution of n-heptane, ethyl acetate and formic acid with the volume ratio of 99:0.5:0.5; the solvent III is alkane and/or ester;
(2) Separating and purifying the I section containing tetrahydrocannabinol and cannabinol and the II section containing cannabinol and cannabinol by column chromatography to obtain primary tetrahydrocannabinol, primary cannabinol and primary cannabinol; the section I and the section II are independently separated and purified by adopting forward column chromatography or reverse column chromatography;
(3) After cracking and destroying the primary tetrahydrocannabinol product, neutral adjustment or inactivation is carried out and then the primary tetrahydrocannabinol product is discarded; concentrating and drying primary cannabinol, primary cannabinol and primary cannabinol respectively.
2. The method for separating cannabinoids as claimed in claim 1, wherein in the step (1), the mass content of tetrahydrocannabinol in the raw material sample is 0.3% or less.
3. The method for separating cannabinoids according to claim 1, wherein in step (1), the raw material sample has a cannabidiol content of 2-20% by mass, a cannabinol content of 1-10% by mass, a cannabichromene content of 0.5-5% by mass, and a cannabinol content of 2-20% by mass.
4. The method according to claim 1, wherein in the step (1), the extracted part of the industrial cannabis extract material is any one or a combination of at least two of cannabis flowers, cannabis leaves, cannabis roots, cannabis stalk cores and cannabis seed meal.
5. The method according to claim 1, wherein in the step (1), the filler used for the solid phase extraction purification comprises any one or a combination of at least two of activated clay, diatomaceous earth, calcium phosphate, calcium carbonate, polyamide powder and silica gel.
6. The method for separating cannabinoids as claimed in claim 5, wherein the particle size of the polyamide powder is 90-120 mesh.
7. The method for separating cannabinoids as claimed in claim 5, wherein the particle size of the silica gel is 100-500 mesh.
8. The method for separating cannabinoids according to claim 1, wherein in the step (1), the mass ratio of the total amount of the filler to the raw material sample in the solid phase extraction column is 1 (0.5-2).
9. The method for separating cannabinoids as claimed in claim 1, wherein in step (1), the solid phase extraction is performed by dry loading or wet loading.
10. The method for separating cannabinoids as claimed in claim 9, characterized in that the dry loading is in particular: mixing part of the filler with the raw material sample to form a solid dispersion system, and transferring the solid dispersion system into a solid phase extraction column filled with the filler; the mass ratio of the partial filler to the raw material sample is 1 (0.25-2).
11. The method for separating cannabinoids as claimed in claim 10, characterized in that the mass ratio of the partial filler to the raw material sample is 1 (0.5-1).
12. The method for separating cannabinoids as claimed in claim 9, characterized in that the wet loading is in particular: mixing an organic solvent with a raw material sample to form a liquid dispersion system, and transferring the liquid dispersion system into a solid phase extraction column filled with a filler; the volume mass ratio of the organic solvent to the raw material sample is 1mL (0.25-5) g.
13. The method for separating cannabinoid according to claim 12, wherein the volume to mass ratio of the organic solvent to the raw material sample is 1mL (0.5-2) g.
14. The method for separating cannabinoids as claimed in claim 9, characterized in that the organic solvent for wet loading is an eluting solvent.
15. The method for separating cannabinoid according to claim 14, wherein the organic solvent for wet loading is any one or a combination of at least two of hexane, heptane or an aqueous ethanol solution having a concentration of 40 to 65 v%.
16. The method for separating cannabinoids as claimed in claim 1, wherein in the step (1), the concentration temperature is 50-65 ℃, and the concentration pressure is-0.10 to-0.065 MPa.
17. The method of claim 1, wherein in step (1), the concentration is to a dry extract having a relative density of 1.05-1.15.
18. The method for separating cannabinoid according to claim 1, wherein the reverse column chromatography is performed using any one of reverse ODS silica gel, polyamide powder or MCI resin powder containing a specific bond.
19. The method of claim 1, wherein the reverse-phase column chromatography eluent is selected from any one or a combination of at least two of alcohol, ketone, ester or water.
20. The method of claim 1, wherein in step (3), the destruction is performed by oxidation, reduction, enzymatic, acid-base cleavage or thermal cleavage.
21. The method for separating cannabinoids as claimed in claim 1, wherein in the step (3), the concentration temperature is 50-65 ℃, and the concentration pressure is-0.10 to-0.065 MPa.
22. The method of claim 1, wherein in step (3), the drying comprises any one or a combination of at least two of vacuum drying, freeze drying, near infrared drying, air drying or microwave drying.
23. The method for separating cannabinoids as claimed in claim 1, wherein in step (3), the drying temperature is 30-65 ℃.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111138400A (en) * 2020-01-20 2020-05-12 云南绿新生物药业有限公司 Extraction and separation method of cannabichromene
CN111315459A (en) * 2017-07-07 2020-06-19 奥罗克姆科技有限公司 Method for the purification and isolation of cannabinoids from dried cannabis sativa and cannabis leaves
WO2020220863A1 (en) * 2019-04-30 2020-11-05 云南汉盟制药有限公司 Method for preparing cannabis sativa l. oil and cannabis sativa l. oil prepared thereby
CN112010738A (en) * 2020-08-05 2020-12-01 晨光生物科技集团股份有限公司 An industrial method for producing cannabinoid compounds by chromatographic separation
WO2020242969A1 (en) * 2019-05-24 2020-12-03 ImMutriX Therapeutics, Inc. Chromatographic media and methods of making and using same

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3199233B1 (en) * 2014-09-25 2018-12-05 Shenzhen Cantonnet Energy Services Co. , Ltd. Preparation method of a solid phase extraction column comprising graphene or modified graphene

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111315459A (en) * 2017-07-07 2020-06-19 奥罗克姆科技有限公司 Method for the purification and isolation of cannabinoids from dried cannabis sativa and cannabis leaves
WO2020220863A1 (en) * 2019-04-30 2020-11-05 云南汉盟制药有限公司 Method for preparing cannabis sativa l. oil and cannabis sativa l. oil prepared thereby
WO2020242969A1 (en) * 2019-05-24 2020-12-03 ImMutriX Therapeutics, Inc. Chromatographic media and methods of making and using same
CN111138400A (en) * 2020-01-20 2020-05-12 云南绿新生物药业有限公司 Extraction and separation method of cannabichromene
CN112010738A (en) * 2020-08-05 2020-12-01 晨光生物科技集团股份有限公司 An industrial method for producing cannabinoid compounds by chromatographic separation

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