Fructus citri sarcodactylis extract and preparation method thereof

Abstract

The invention relates to the technical field of plant extraction, in particular to a fingered citron extract and a preparation method thereof.The preparation method comprises the steps that fingered citron fruits are smashed and then subjected to wet granulation pretreatment, and fingered citron particles are formed; extracting the fingered citron particles by adopting supercritical carbon dioxide to obtain extract liquor; carrying out clathration treatment on the extraction liquid by adopting cyclodextrin to form clathration liquid; and carrying out vacuum concentration, cooling standing and centrifugal treatment on the inclusion liquid, taking a precipitate after centrifugation, and drying to obtain the fingered citron extract. According to the preparation method, the content and the transfer rate of nomilin in the fingered citron extract can be effectively improved, and the content and the yield of nomilin in the prepared fingered citron extract are higher.

Description

Technical Field

[0001] The invention relates to the technical field of plant extraction, in particular to a bergamot extract and a preparation method thereof. Background Art

[0002] Buddha's hand is an evergreen shrub or small tree of the genus Citrus in the Rutaceae family. It is a variant of citron (Citrus medica L.). Buddha's hand has a long history of cultivation in China and is mainly planted in Guangdong, Sichuan, Zhejiang and Fujian. In addition, it is also cultivated in Guangxi, Anhui, Yunnan and other places. Due to different origins, it is mainly divided into varieties such as Guangdong Buddha's hand, Sichuan Buddha's hand, Golden Buddha's hand and Jian Buddha's hand. The fruit of Buddha's hand tastes spicy, bitter, sour, warm in nature, and non-toxic; it enters the spleen, liver, stomach, and lung meridians, and has multiple functions such as regulating qi and resolving phlegm, stopping vomiting and relieving swelling, soothing the liver and strengthening the spleen, and harmonizing the stomach and relieving pain. As a traditional Chinese medicinal material in my country, Buddha's hand has extremely high medicinal value and can be used in combination with a variety of Chinese medicinal materials. It has a good effect on diseases such as liver and stomach qi stagnation, chest and flank distension and pain, stomach fullness, poor appetite and vomiting, cough and sputum. Modern medical research has proved that Buddha's hand has anti-depressant, antibacterial, anti-inflammatory, anti-cancer, anti-tumor, anti-aging, and blood pressure lowering effects.

[0003] At present, many chemical components have been extracted from the fruit of the medicinal part of Buddha's hand, which can be roughly divided into the following categories: polysaccharides, flavonoids, limonoids, polyphenols, etc. Among them, Nomilin is a limonoid compound. Nomilin has a complex structure and is expensive because its content in natural plants is generally low. The market price is as high as more than 20,000. At present, there are many studies on limonoids, but few studies on Nomilin. Among them, the method of extracting Nomilin from natural plants is mainly to directly crush the citrus seeds such as honey pomelo, kumquat, sour orange or lemon peel residue and other plant powder materials, and then extract them by solution reflux extraction, supercritical extraction or ultrasonic extraction. However, the extracts obtained by these extraction methods contain more limonoids, and the purity of the Nomilin they contain is low and the yield is low.

[0004] It has been reported that limonoid compounds can be extracted from bergamot. Among them, the alcohol extract, acetone extract and ethyl acetate extract of bergamot mostly contain limonoids and flavonoids, and the purity of the nomilin contained therein is low and the yield is low. Summary of the invention

[0005] The purpose of the present invention is to provide a bergamot extract and a preparation method thereof in view of the existing technical status.

[0006] The present invention performs wet granulation pre-treatment on crushed bergamot fruits, and then extracts the obtained bergamot particles using supercritical carbon dioxide technology to better replace the fat-soluble component Nomilin. Subsequently, cyclodextrin is used to carry out inclusion treatment on the extract enriched with Nomilin, and the inclusion liquid is reduced-pressure concentrated and cooled and allowed to stand, so that subsequent centrifugal treatment can separate more Nomilin inclusion compounds, thereby effectively improving the content and transfer rate of Nomilin in the bergamot extract, and the content and yield of Nomilin in the obtained bergamot extract are higher.

[0007] In order to achieve the above object, the present invention adopts the following technical scheme:

[0008] First, the present invention provides a method for preparing a bergamot extract, comprising:

[0009] After the bergamot fruit is crushed, it is subjected to wet granulation pre-treatment to form bergamot granules;

[0010] Extracting the bergamot particles with supercritical carbon dioxide to obtain an extract;

[0011] The extract is subjected to inclusion treatment by using cyclodextrin to form an inclusion liquid;

[0012] The inclusion solution is concentrated under reduced pressure, cooled and allowed to stand, and centrifuged. After centrifugation, the precipitate is taken and dried to obtain the bergamot extract.

[0013] In some embodiments, the step of pre-treatment before wet granulation comprises:

[0014] The crushed bergamot fruit is used as a powder material raw material, and an ethanol solution is used as a binder to prepare a soft material, and then the soft material is made into wet granules, and the wet granules are low-temperature dried and granulated to form the bergamot granules.

[0015] In some embodiments, the ethanol solution is 50% to 95% ethanol, and the weight ratio between the powder raw material and the adhesive is 1:0.5 to 2.

[0016] In some embodiments, the particle size of the bergamot particles is 30 mesh to 80 mesh.

[0017] In some embodiments, the step of using cyclodextrin to include the extract comprises:

[0018] dissolving cyclodextrin in water to prepare a saturated cyclodextrin aqueous solution;

[0019] A saturated cyclodextrin aqueous solution is added to the extract while stirring, followed by high-speed shearing with a shearing machine or a colloid mill to form the inclusion liquid.

[0020] In some embodiments, the cyclodextrin is one or more combinations of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

[0021] In some embodiments, after the reduced pressure concentration, high-speed shearing is performed using a shearing machine or a colloid mill, followed by cooling and standing; in the reduced pressure concentration step, the concentration temperature is controlled below 55° C., and the concentrate is concentrated to a solid content mass fraction greater than 10%.

[0022] In some embodiments, in the cooling and standing step, the cooling and standing time is more than 8 hours, and the cooling and standing temperature is 0°C to 8°C.

[0023] In some embodiments, in the step of extracting the bergamot particles with supercritical carbon dioxide, the extraction time is 1h to 3h, the extraction temperature is 40°C to 50°C, the extraction pressure is 20MPa to 30MPa, the entraining agent is ethanol, and the amount of the entraining agent is 2 to 3 times the weight of the bergamot particles.

[0024] Secondly, the present invention also provides a bergamot extract, which is prepared by the above-mentioned preparation method of the bergamot extract.

[0025] The beneficial effects of the present invention are:

[0026] In the present invention, first, after the bergamot fruit is powdered, wet granulation is performed to prepare granular bergamot particles, and then the bergamot particles are subjected to supercritical carbon dioxide extraction. Compared with powder, the bergamot particles can be relatively loosely accumulated in the extraction kettle, so that during the supercritical extraction process, the bergamot raw material can be fully contacted with the carbon dioxide extractant, and the carbon dioxide extractant can better infiltrate the bergamot raw material cells during the high-pressure flow process, thereby better replacing the fat-soluble component Nomilin; in addition, the bergamot raw material contains a large amount of water-soluble substances, such as polysaccharides, proteins, polyphenols, etc., and Nomilin is a small molecule fat-soluble substance. The extraction of flavonoids and limonoid fat-soluble substances by supercritical carbon dioxide can greatly enrich the effective components of Nomilin, thereby effectively improving the yield. Subsequently, cyclodextrin was used to carry out inclusion treatment on the extract enriched with Nomilin. Nomilin molecules contain more ester bonds and methyl groups, which are easy to form hydrogen bonds and van der Waals forces with the hydrophobic cavity in cyclodextrin. Therefore, the inclusion of Nomilin with cyclodextrin can not only significantly improve the purity, but also promote its solubility in the aqueous phase. Since the inclusion treatment was carried out first and then the reduced pressure concentration was carried out, the inclusion effect of cyclodextrin can effectively prevent the thermal decomposition of the heat-sensitive substance Nomilin from being heated for too long, and the inclusion liquid was reduced pressure and concentrated again, and precipitation was continuously precipitated during the concentration process. At the same time, the purity of the volatile solvent was reduced, which prompted the fat-soluble Nomilin to better enter the hydrophobic cyclodextrin cavity, which can improve the effect of inclusion separation. After reduced pressure concentration, the solubility of the inclusion compound in the inclusion liquid was reduced by cooling and standing, and the inclusion compound was more likely to aggregate or precipitate between molecules at low temperatures, which was beneficial to the subsequent centrifugal treatment, thereby effectively improving the content and transfer rate of Nomilin.

[0027] Therefore, the present invention performs wet granulation pre-treatment on the crushed bergamot fruit, and then extracts the obtained bergamot particles using supercritical carbon dioxide technology to better replace the fat-soluble component Nomilin. Subsequently, cyclodextrin is used to carry out inclusion treatment on the extract enriched with Nomilin, and the inclusion liquid is reduced pressure, concentrated and cooled to stand, so that subsequent centrifugation can separate more Nomilin inclusion complexes, thereby effectively improving the content and transfer rate of Nomilin in the bergamot extract. The content of Nomilin in the obtained bergamot extract can be as high as 8% or more, and the yield can be as high as 60% or more. Compared with the existing process for extracting Nomilin from plants, the content and yield of Nomilin are higher. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The present invention is a flow chart of a method for preparing a bergamot extract. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention is described in further detail below.

[0030] First, see Figure 1 As shown, the present invention provides a method for preparing a bergamot extract, comprising:

[0031] S100. After the bergamot fruit is crushed, it is subjected to wet granulation pre-treatment to form bergamot granules;

[0032] S200. extracting the bergamot particles using supercritical carbon dioxide to obtain an extract;

[0033] S300. The extract is subjected to inclusion treatment using cyclodextrin to form an inclusion liquid;

[0034] S400. The inclusion solution is concentrated under reduced pressure, cooled and allowed to stand, and centrifuged. After centrifugation, the precipitate is collected and dried to obtain a bergamot extract.

[0035] In the present invention, first, after the bergamot fruit is powdered, wet granulation is performed to prepare granular bergamot particles, and then the bergamot particles are subjected to supercritical carbon dioxide extraction. Compared with powder, the bergamot particles can be relatively loosely accumulated in the extraction kettle, so that during the supercritical extraction process, the bergamot raw material can be fully contacted with the carbon dioxide extractant, and the carbon dioxide extractant can better infiltrate the bergamot raw material cells during the high-pressure flow process, thereby better replacing the fat-soluble component Nomilin; in addition, the bergamot raw material contains a large amount of water-soluble substances, such as polysaccharides, proteins, polyphenols, etc., and Nomilin is a small molecule fat-soluble substance. The extraction of flavonoids and limonoid fat-soluble substances by supercritical carbon dioxide can greatly enrich the effective components of Nomilin, thereby effectively improving the yield. Subsequently, cyclodextrin was used to carry out inclusion treatment on the extract enriched with Nomilin. Nomilin molecules contain more ester bonds and methyl groups, which are easy to form hydrogen bonds and van der Waals forces with the hydrophobic cavity in cyclodextrin. Therefore, the inclusion of Nomilin with cyclodextrin can not only significantly improve the purity, but also promote its solubility in the aqueous phase. Since the inclusion treatment was carried out first and then the reduced pressure concentration was carried out, the inclusion effect of cyclodextrin can effectively prevent the thermal decomposition of the heat-sensitive substance Nomilin from being heated for too long, and the inclusion liquid was reduced pressure and concentrated again, and precipitation was continuously precipitated during the concentration process. At the same time, the purity of the volatile solvent was reduced, which prompted the fat-soluble Nomilin to better enter the hydrophobic cyclodextrin cavity, which can improve the effect of inclusion separation. After reduced pressure concentration, the solubility of the inclusion compound in the inclusion liquid was reduced by cooling and standing, and the inclusion compound was more likely to aggregate or precipitate between molecules at low temperatures, which was beneficial to the subsequent centrifugal treatment, thereby effectively improving the content and transfer rate of Nomilin.

[0036] Therefore, the present invention performs wet granulation pre-treatment on the crushed bergamot fruit, and then extracts the obtained bergamot particles using supercritical carbon dioxide technology to better replace the fat-soluble component Nomilin. Subsequently, cyclodextrin is used to carry out inclusion treatment on the extract enriched with Nomilin, and the inclusion liquid is reduced pressure, concentrated and cooled to stand, so that subsequent centrifugation can separate more Nomilin inclusion complexes, thereby effectively improving the content and transfer rate of Nomilin in the bergamot extract. The content of Nomilin in the obtained bergamot extract can be as high as 8% or more, and the yield can be as high as 60% or more. Compared with the existing process for extracting Nomilin from plants, the content and yield of Nomilin are higher.

[0037] In addition, the nomilin in the prepared product is included, and the performance is more stable, which is conducive to subsequent applications.

[0038] The bergamot extract prepared by the invention contains high content of nomilin, is soluble in water, and can be applied to health food for preventing pancreatic cancer, liver cancer, bladder cancer and other cancers, or to the preparation of common anti-oxidative food.

[0039] In some embodiments, in step S100, the wet granulation pre-treatment step includes:

[0040] The crushed bergamot fruit is used as a powder material raw material, and an ethanol solution is used as a binder to prepare a soft material, and then the soft material is made into wet granules, and the wet granules are low-temperature dried and granulated to form the bergamot granules.

[0041] In the wet granulation process, ethanol solution is used as a binder, which can not only ensure that the material is dried at a low temperature and protect nomilin from being easily oxidized, but also obtain a relatively loose granular material. In the process of supercritical extraction, the bergamot particles can not only be loosely stacked in the extraction kettle, but the bergamot particles themselves are also relatively loose, ensuring that the bergamot raw material is in more complete contact with the carbon dioxide extractant. During the high-pressure flow process, the carbon dioxide extractant can better infiltrate into the bergamot raw material cells, thereby better replacing the fat-soluble component nomilin, effectively improving the yield.

[0042] In some embodiments, the bergamot fruit is crushed and sieved, and then subjected to a pre-wet granulation treatment.

[0043] In some embodiments, in the step of pre-treatment of wet granulation, the low-temperature drying is fluidized bed drying below 50°C, and the moisture content of the wet granules is below 10%. Low-temperature drying can effectively avoid thermal decomposition of the heat-sensitive material Nomilin during the drying process.

[0044] In some embodiments, the ethanol solution is ethanol with a volume fraction of 50% or more.

[0045] In some embodiments, preferably, the ethanol solution is 50% to 95% ethanol, and the weight ratio between the powder material and the adhesive is 1:0.5 to 2.

[0046] Using a higher concentration of ethanol solution as a binder is more conducive to ensuring that the material can be dried at a low temperature, protecting the nomilin from being easily oxidized, and at the same time obtaining a looser granular material, further improving the yield.

[0047] Exemplarily, the weight ratio between the powder raw material and the adhesive is 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2, but is not limited thereto.

[0048] In some embodiments, the particle size of the bergamot particles is 30 to 80 meshes. Within this particle size range, during the supercritical extraction process, the bergamot particles can be more fully contacted with the carbon dioxide extractant, and the fat-soluble component Nomilin can be better replaced, further improving the yield. When the particle size of the bergamot particles is too large, the inside of the bergamot particles is not easy to dry and is not easy to fully contact with the carbon dioxide extractant.

[0049] In some embodiments, in the step of pre-treatment of wet granulation, the granules are sieved through a 30-mesh sieve and then a 80-mesh sieve.

[0050] In some embodiments, in step S300, the step of using cyclodextrin to include the extract comprises:

[0051] S310. Dissolving cyclodextrin in water to prepare a saturated cyclodextrin aqueous solution;

[0052] S320. Adding a saturated cyclodextrin aqueous solution to the extract while stirring, followed by high-speed shearing with a shearing machine or a colloid mill to form the inclusion liquid.

[0053] After the saturated cyclodextrin aqueous solution is mixed with the extract, it is subjected to high-speed shearing by a shearing machine or a colloid mill. The high-speed shearing method breaks the agglomeration between the molecules, increases the frequency of collision between the two molecules, thereby increasing the chances of contact and inclusion of cyclodextrin and nomiline, and improving the inclusion separation effect.

[0054] In some embodiments, the high-speed shearing treatment time of the shearing machine or colloid mill is 10 minutes to 30 minutes to increase the chance of contact and inclusion of cyclodextrin and nomiline. Preferably, the high-speed shearing treatment time of the shearing machine or colloid mill is 15 minutes.

[0055] In some embodiments, in step S310, water is first heated to 45°C to 55°C, and cyclodextrin is added under stirring conditions to prepare a saturated cyclodextrin aqueous solution. The water temperature should not be too high to avoid subsequent mixing with the extract causing thermal decomposition of nomiline. Exemplarily, the water is heated to 50°C.

[0056] In some embodiments, the cyclodextrin is one or more combinations of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin.

[0057] Preferably, the cyclodextrin is β-cyclodextrin. Compared with other types of cyclodextrins, β-cyclodextrin has a cavity and solubility that are more suitable for inclusion of nomiline, so that β-cyclodextrin can obtain a better inclusion and separation effect.

[0058] In some embodiments, after the reduced pressure concentration, high-speed shearing is performed using a shearing machine or a colloid mill, followed by cooling and standing; in the reduced pressure concentration step, the concentration temperature is controlled below 55° C., and the concentrate is concentrated to a solid content mass fraction greater than 10%.

[0059] After the inclusion treatment, the inclusion liquid is subjected to reduced pressure concentration and high-speed shearing to break the intermolecular agglomeration, increase the collision frequency between the two molecules, thereby improving the chance of contact and inclusion, and at the same time reduce the purity of ethanol, so as to promote the fat-soluble nomilin to better enter the hydrophobic cyclodextrin cavity, further improving the inclusion separation effect; in the reduced pressure concentration step, the concentration temperature is controlled below 55°C to avoid thermal decomposition of the heat-sensitive substance nomilin, thereby further increasing the content and transfer rate of nomilin in the bergamot extract.

[0060] In some embodiments, in the cooling and standing step, the cooling and standing time is more than 8 hours, and the cooling and standing temperature is 0°C to 8°C.

[0061] Illustratively, the cooling and standing temperature is 0° C., 2° C., 4° C. or 8° C., but is not limited thereto.

[0062] By means of cooling and standing, the solubility of the inclusion compound in the inclusion solution is reduced, and the inclusion compound is more likely to aggregate or precipitate molecules at low temperatures, thereby increasing the transfer rate of Nomilin.

[0063] In some embodiments, in the step of extracting the bergamot particles with supercritical carbon dioxide, the extraction time is 1h to 3h, the extraction temperature is 40°C to 50°C, the extraction pressure is 20MPa to 30MPa, the entraining agent is ethanol, and the amount of the entraining agent is 2 to 3 times the weight of the bergamot particles.

[0064] After wet granulation, supercritical carbon dioxide extraction is carried out. The granulated bergamot particles are extracted and separated using carbon dioxide fluid under low temperature conditions, and ethanol is used as an entrainer. This can better replace the fat-soluble component nomilin, effectively increasing the content and transfer rate of nomilin in the bergamot extract.

[0065] In some embodiments, the precipitate is dried by freeze drying.

[0066] Through freeze-drying, the thermal decomposition of nomilin during the drying process can be effectively avoided, and the content and transfer rate of nomilin in bergamot extract can be further increased.

[0067] Preferably, the freeze-drying step comprises:

[0068] The precipitate was pre-frozen at -50°C for more than 6 hours, vacuumized, and then subjected to programmed cooling and sublimation drying. After drying, the freeze-dried powder was crushed to obtain a bergamot extract.

[0069] Secondly, the present invention also provides a bergamot extract, which is prepared by the above-mentioned preparation method of the bergamot extract.

[0070] The present invention will be further described below in conjunction with the accompanying drawings and embodiments:

[0071] Example 1

[0072] This embodiment discloses a method for preparing a bergamot extract, comprising:

[0073] S100. After the bergamot fruit is crushed, it is subjected to wet granulation pre-treatment to form bergamot granules:

[0074] S111. Screening 2kg of bergamot fruit that is not moldy, crushing the bergamot fruit and passing it through a 60-mesh sieve;

[0075] S112. Use crushed bergamot fruit as powder material and 75% ethanol solution as adhesive, with the weight ratio of the powder material to the adhesive being 1:1.5, to prepare soft material, extrude and form, pass through a 30-mesh sieve, make the soft material into wet granules, dry the wet granules in a fluidized bed below 50°C until the moisture content of the wet granules is below 10%, and pass through a 30-mesh sieve and an 80-mesh sieve to form bergamot granules.

[0076] S200. Extract the bergamot particles with supercritical carbon dioxide to obtain an extract:

[0077] The bergamot particles were placed in an extraction kettle with an extraction temperature of 40°C, an extraction pressure of 30MPa, a separation kettle pressure of 5Mpa, and a carbon dioxide extractant flow rate of 20L / h. After the parameters were stable, ethanol was added as an entrainer with an ethanol flow rate of 2L / h. The extraction was started for 2h. The amount of the entrainer was twice the weight of the bergamot particles. The extract was collected from the separation kettle during the extraction process and combined after the extraction was completed.

[0078] S300. The extract is subjected to inclusion treatment using cyclodextrin to form an inclusion solution:

[0079] S310. Weigh cyclodextrin according to the weight of 50% of the dry paste of the extract, dissolve the cyclodextrin in water at 50°C to prepare a saturated cyclodextrin aqueous solution, wherein the cyclodextrin is α-cyclodextrin;

[0080] S320. Add a saturated cyclodextrin aqueous solution to the extract while stirring, and then perform high-speed shearing with a shearing machine or a colloid mill for 15 minutes to form the inclusion liquid.

[0081] S400. The inclusion solution is concentrated under reduced pressure, cooled, allowed to stand and centrifuged, and the precipitate is taken after centrifugation and dried to obtain a bergamot extract:

[0082] The inclusion liquid is concentrated under reduced pressure, and the concentration temperature is controlled below 55° C., and concentrated to a concentrated liquid with a solid content mass fraction greater than 10%. A shearing machine or a colloid mill is used for high-speed shearing for 15 minutes, and then the liquid is transferred to a cold storage at 4° C. and refrigerated for 10 hours. The precipitate is obtained by centrifugation, and the precipitate is pre-frozen at minus 50° C. for 10 hours, vacuumed, and then programmed cooling and sublimation drying is performed. After drying, the freeze-dried powder is taken and crushed to obtain a bergamot extract.

[0083] This embodiment also discloses a bergamot extract, which is prepared by the above-mentioned preparation method of the bergamot extract.

[0084] Example 2

[0085] This embodiment discloses a method for preparing a bergamot extract, comprising:

[0086] S100. After the bergamot fruit is crushed, it is subjected to wet granulation pre-treatment to form bergamot granules:

[0087] S111. Screening 2kg of bergamot fruit that is not moldy, crushing the bergamot fruit and passing it through a 60-mesh sieve;

[0088] S112. Use crushed bergamot fruit as powder material and 75% ethanol solution as adhesive, with the weight ratio of the powder material to the adhesive being 1:1.5, to prepare soft material, extrude and form, pass through a 30-mesh sieve, make the soft material into wet granules, dry the wet granules in a fluidized bed below 50°C until the moisture content of the wet granules is below 10%, and pass through a 30-mesh sieve and an 80-mesh sieve to form bergamot granules.

[0089] S200. Extract the bergamot particles with supercritical carbon dioxide to obtain an extract:

[0090] The bergamot particles were placed in an extraction kettle with an extraction temperature of 40°C, an extraction pressure of 30MPa, a separation kettle pressure of 5Mpa, and a carbon dioxide extractant flow rate of 20L / h. After the parameters were stable, ethanol was added as an entrainer with an ethanol flow rate of 2L / h. The extraction was started for 2h. The amount of the entrainer was twice the weight of the bergamot particles. The extract was collected from the separation kettle during the extraction process and combined after the extraction was completed.

[0091] S300. The extract is subjected to inclusion treatment using cyclodextrin to form an inclusion solution:

[0092] S310. Weigh cyclodextrin according to the weight of 50% of the dry paste of the extract, dissolve the cyclodextrin in water at 50°C to prepare a saturated cyclodextrin aqueous solution, wherein the cyclodextrin is β-cyclodextrin;

[0093] S320. Add a saturated cyclodextrin aqueous solution to the extract while stirring, and then perform high-speed shearing with a shearing machine or a colloid mill for 15 minutes to form the inclusion liquid.

[0094] S400. The inclusion solution is concentrated under reduced pressure, cooled, allowed to stand and centrifuged, and the precipitate is taken after centrifugation and dried to obtain a bergamot extract:

[0095] The inclusion liquid is concentrated under reduced pressure, and the concentration temperature is controlled below 55° C., and concentrated to a concentrated liquid with a solid content mass fraction greater than 10%. A shearing machine or a colloid mill is used for high-speed shearing for 15 minutes, and then the liquid is transferred to a cold storage at 4° C. and refrigerated for 10 hours. The precipitate is obtained by centrifugation, and the precipitate is pre-frozen at minus 50° C. for 10 hours, vacuumed, and then programmed cooling and sublimation drying is performed. After drying, the freeze-dried powder is taken and crushed to obtain a bergamot extract.

[0096] This embodiment also discloses a bergamot extract, which is prepared by the above-mentioned preparation method of the bergamot extract.

[0097] Example 3

[0098] This embodiment discloses a method for preparing a bergamot extract, comprising:

[0099] S100. After the bergamot fruit is crushed, it is subjected to wet granulation pre-treatment to form bergamot granules:

[0100] S111. Screening 2kg of bergamot fruit that is not moldy, crushing the bergamot fruit and passing it through a 60-mesh sieve;

[0101] S112. Use crushed bergamot fruit as powder material and 50% ethanol solution as adhesive, with the weight ratio of the powder material to the adhesive being 1:1.5, to prepare a soft material, extrude it into shape, pass it through a 30-mesh sieve, make the soft material into wet granules, dry the wet granules in a fluidized bed below 50°C until the moisture content of the wet granules is below 10%, and pass it through a 30-mesh sieve and an 80-mesh sieve to form the bergamot granules.

[0102] S200. Extract the bergamot particles with supercritical carbon dioxide to obtain an extract:

[0103] The bergamot particles were placed in an extraction kettle with an extraction temperature of 40°C, an extraction pressure of 30MPa, a separation kettle pressure of 5Mpa, and a carbon dioxide extractant flow rate of 20L / h. After the parameters were stable, ethanol was added as an entrainer with an ethanol flow rate of 2L / h. The extraction was started for 2h. The amount of the entrainer was twice the weight of the bergamot particles. The extract was collected from the separation kettle during the extraction process and combined after the extraction was completed.

[0104] S300. The extract is subjected to inclusion treatment using cyclodextrin to form an inclusion solution:

[0105] S310. Weigh cyclodextrin according to the weight of 50% of the dry paste of the extract, dissolve the cyclodextrin in water at 50°C to prepare a saturated cyclodextrin aqueous solution, wherein the cyclodextrin is γ-cyclodextrin;

[0106] S320. Add a saturated cyclodextrin aqueous solution to the extract while stirring, and then perform high-speed shearing with a shearing machine or a colloid mill for 15 minutes to form the inclusion liquid.

[0107] S400. The inclusion solution is concentrated under reduced pressure, cooled, allowed to stand and centrifuged, and the precipitate is taken after centrifugation and dried to obtain a bergamot extract:

[0108] The inclusion liquid is concentrated under reduced pressure, and the concentration temperature is controlled below 55° C., and concentrated to a concentrated liquid with a solid content mass fraction greater than 10%. A shearing machine or a colloid mill is used for high-speed shearing for 15 minutes, and then the liquid is transferred to a cold storage at 4° C. and refrigerated for 10 hours. The precipitate is obtained by centrifugation, and the precipitate is pre-frozen at minus 50° C. for 10 hours, vacuumed, and then programmed cooling and sublimation drying is performed. After drying, the freeze-dried powder is taken and crushed to obtain a bergamot extract.

[0109] This embodiment also discloses a bergamot extract, which is prepared by the above-mentioned preparation method of the bergamot extract.

[0110] Comparative Example 1

[0111] This comparative example discloses a method for preparing a bergamot extract, comprising:

[0112] S11. Screen 2kg of mildew-free materials and crush them into 20-mesh particles.

[0113] S12. Place the particles in a water bath reflux extractor, add 95% ethanol, and reflux extract twice. The first time, add 10 times the ethanol and reflux extract for 2 hours, and the second time, add 8 times the ethanol and reflux extract for 2.0 hours.

[0114] S13. The extract is filtered, and the filtrate is concentrated under reduced pressure. The concentration temperature is controlled below 55° C. and the concentration is performed until there is no obvious ethanol smell to obtain a concentrated solution.

[0115] S14. The concentrated solution is subjected to inclusion treatment using cyclodextrin to form an inclusion solution:

[0116] S141. Weigh cyclodextrin according to the weight of 50% of the dry paste of the concentrate, dissolve the cyclodextrin in water at 50°C to prepare a saturated cyclodextrin aqueous solution, wherein the cyclodextrin is β-cyclodextrin;

[0117] S142. Add a saturated cyclodextrin aqueous solution to the concentrated solution while stirring, and then perform high-speed shearing with a shearing machine or a colloid mill for 15 minutes to form the inclusion liquid.

[0118] S5. The inclusion solution is concentrated under reduced pressure, cooled, allowed to stand and centrifuged, and the precipitate is taken after centrifugation and dried to obtain a bergamot extract:

[0119] The inclusion liquid is concentrated under reduced pressure, and the concentration temperature is controlled below 55° C., and concentrated to a concentrated liquid with a solid content mass fraction greater than 10%. A shearing machine or a colloid mill is used for high-speed shearing for 15 minutes, and then the liquid is transferred to a cold storage at 4° C. and refrigerated for 10 hours. The precipitate is obtained by centrifugation, and the precipitate is pre-frozen at minus 50° C. for 10 hours, vacuumed, and then programmed cooling and sublimation drying is performed. After drying, the freeze-dried powder is taken and crushed to obtain a bergamot extract.

[0120] This embodiment also discloses a bergamot extract, which is prepared by the above-mentioned preparation method of the bergamot extract.

[0121] Comparative Example 2

[0122] This comparative example discloses a method for preparing a bergamot extract, comprising:

[0123] S21. Screen 2kg of mildew-free materials and crush them into 30-mesh particles.

[0124] S22. Extract the particles using supercritical carbon dioxide to obtain an extract:

[0125] The particles were placed in an extraction kettle with an extraction temperature of 40°C, an extraction pressure of 30MPa, a separation kettle pressure of 5Mpa, and a carbon dioxide extractant flow rate of 20L / h. After the parameters were stable, ethanol was added as an entrainer with an ethanol flow rate of 2L / h. The extraction was started for 2h. The amount of entrainer was twice the weight of the bergamot particles. The extract was collected from the separation kettle during the extraction process and combined after the extraction was completed.

[0126] S23. Take the extract and concentrate under reduced pressure and dry it:

[0127] S231. The extract is concentrated under reduced pressure, the concentration temperature is controlled below 55 ° C, and concentrated to a concentrated solution having a solid content of greater than 10% by mass to obtain a concentrated solution;

[0128] S232. Pre-freeze the concentrated solution at -50°C for 10 hours, evacuate the solution, and then perform programmed cooling and sublimation drying. After drying, take the freeze-dried powder and grind it to obtain the bergamot extract.

[0129] Comparative Example 3

[0130] S31. Screen 2 kg of mildew-free materials and crush them through a 30-mesh sieve.

[0131] S32. Extracting the particles with supercritical carbon dioxide to obtain an extract:

[0132] The particles were placed in an extraction kettle with an extraction temperature of 40°C, an extraction pressure of 30MPa, a separation kettle pressure of 5Mpa, and a carbon dioxide extractant flow rate of 20L / h. After the parameters were stable, ethanol was added as an entrainer with an ethanol flow rate of 2L / h. The extraction was started for 2h. The amount of entrainer was twice the weight of the bergamot particles. The extract was collected from the separation kettle during the extraction process and combined after the extraction was completed.

[0133] S33. The extract is subjected to inclusion treatment using cyclodextrin to form an inclusion solution:

[0134] S331. Weigh cyclodextrin according to the weight of 50% of the dry paste of the extract, dissolve the cyclodextrin in water at 50°C to prepare a saturated cyclodextrin aqueous solution, wherein the cyclodextrin is β-cyclodextrin;

[0135] S332. Add a saturated cyclodextrin aqueous solution to the extract while stirring, and then perform high-speed shearing with a shearing machine or a colloid mill for 15 minutes to form the inclusion liquid.

[0136] S34. The inclusion solution is concentrated under reduced pressure, cooled, allowed to stand and centrifuged, and the precipitate is taken after centrifugation and dried to obtain a bergamot extract:

[0137] The inclusion liquid is concentrated under reduced pressure, and the concentration temperature is controlled below 55° C., and concentrated to a concentrated liquid with a solid content mass fraction greater than 10%. A shearing machine or a colloid mill is used for high-speed shearing for 15 minutes, and then the liquid is transferred to a cold storage at 4° C. and refrigerated for 10 hours. The precipitate is obtained by centrifugation, and the precipitate is pre-frozen at minus 50° C. for 10 hours, vacuumed, and then programmed cooling and sublimation drying is performed. After drying, the freeze-dried powder is taken and crushed to obtain a bergamot extract.

[0138] Comparative Example 4

[0139] S41. Screen 2 kg of mildew-free material and crush through a 30-mesh sieve.

[0140] S42. Extracting the particles with supercritical carbon dioxide to obtain an extract:

[0141] The particles were placed in an extraction kettle with an extraction temperature of 40°C, an extraction pressure of 30MPa, a separation kettle pressure of 5Mpa, and a carbon dioxide extractant flow rate of 20L / h. After the parameters were stable, ethanol was added as an entrainer with an ethanol flow rate of 2L / h. The extraction was started for 2h. The amount of entrainer was twice the weight of the bergamot particles. The extract was collected from the separation kettle during the extraction process and combined after the extraction was completed.

[0142] S43. The extract is subjected to inclusion treatment using cyclodextrin to form an inclusion solution:

[0143] S431. Weigh cyclodextrin according to the weight of 50% of the dry paste of the extract, dissolve the cyclodextrin in water at 50°C to prepare a saturated cyclodextrin aqueous solution, wherein the cyclodextrin is β-cyclodextrin;

[0144] S432. Add a saturated cyclodextrin aqueous solution to the extract while stirring, and then perform high-speed shearing with a shearing machine or a colloid mill for 15 minutes to form the inclusion liquid.

[0145] S44. The inclusion solution is centrifuged, and the precipitate is taken after centrifugation and dried to obtain a bergamot extract:

[0146] The inclusion liquid is centrifuged to obtain a precipitate, which is pre-frozen at -50°C for 10 hours, evacuated, and then subjected to programmed cooling and sublimation drying. After drying, the freeze-dried powder is pulverized to obtain a bergamot extract.

[0147] Nomilin content and transfer rate detection:

[0148] 1. Experimental group: Example 1 to Example 3, Comparative Example 1 to Comparative Example 4;

[0149] 2. Experimental methods:

[0150] 2.1. Preparation of reference solution: Take an appropriate amount of Nomilin reference substance, weigh it accurately, and add methanol to make a solution containing 50 μg per 1 ml.

[0151] 2.2. Take 3g of the lyophilized powder prepared in each experimental group, accurately weigh it, put it in a stoppered conical flask, accurately add 25ml of methanol, seal it, weigh it, ultrasonically treat it for 30 minutes, let it cool, quantify it, accurately pipette 10μl of the reference solution and the test solution, and inject it into a liquid chromatograph, using octadecylsilane bonded silica gel as the filler (Agilent Eclipse XDB-C18250mm, 5μm, 4.6×250mm), acetonitrile as the mobile phase A, and 0.1% phosphoric acid solution as the mobile phase B. Gradient elution was performed according to the provisions in Table 1 below, with a flow rate of 0.8ml (medicinal material) per minute (lyophilized powder / concentrate: 0.6ml), a column temperature of 35°C, and a detection wavelength of 210nm.

[0152] Table 1 Gradient elution table

[0153]

[0154] 3. The experimental results are shown in Table 2 below:

[0155] Table 2 Nomilin content and transfer rate of each experimental group

[0156]

[0157]

[0158] It can be seen from the experimental results that the nomilin content and transfer rate of Examples 1 to 3 of the present invention are better than those of the comparative example. It can be seen that the embodiments of the present invention use wet granulation, supercritical extraction, inclusion treatment, reduced pressure concentration, cooling and standing to separate the crushed bergamot fruit to obtain a bergamot extract with a higher nomilin content.

[0159] Among them, by comparing Examples 1 to 3, it can be seen that β-cyclodextrin has a cavity and solubility that is more suitable for inclusion of nomiline than α-cyclodextrin and γ-cyclodextrin.

[0160] Compared with Comparative Example 3, Example 2 adds a pre-treatment before wet granulation, and the content of Nomilin is increased by 60.8%, and the transfer rate is increased by 52.3%. After wet granulation, the contact area of ​​the bergamot raw material during supercritical extraction is increased, so that the carbon dioxide extractant can better infiltrate into the bergamot raw material cells during the high-pressure flow process, and then the fat-soluble component Nomilin is better replaced. In the wet granulation, a higher concentration of ethanol is used as a binder, which is more conducive to drying the medicinal material particles at low temperature and protecting Nomilin from oxidation.

[0161] It can be seen from Example 2, Comparative Example 1 and Comparative Example 2 that, compared with conventional water bath alcohol extraction, the use of supercritical carbon dioxide extraction and cyclodextrin inclusion separation can significantly increase the content and transfer rate of nomilin in bergamot extract.

[0162] It can be seen from Example 2, Comparative Example 3 and Comparative Example 4 that in the cyclodextrin inclusion process, the inclusion liquid is again reduced in pressure and concentrated, and high-speed shearing is used to break the intermolecular agglomeration, so that the collision frequency between the two molecules is increased, thereby improving the chance of contact and inclusion, and at the same time, the ethanol purity is reduced, which promotes the fat-soluble nomilin to better enter the hydrophobic cyclodextrin cavity; then the solution is cooled and allowed to stand before centrifugation, and the solubility of the inclusion complex is reduced during the cooling process. The inclusion complex is more prone to intermolecular aggregation or precipitation at low temperatures, thereby significantly improving the content and transfer rate of nomilin.

[0163] In summary, the present invention performs wet granulation pre-treatment on the crushed bergamot fruit, and then extracts the obtained bergamot particles using supercritical carbon dioxide technology to better replace the fat-soluble component nomilin. Subsequently, cyclodextrin is used to include the extract enriched with nomilin, and the inclusion liquid is reduced pressure, concentrated, cooled and allowed to stand, so that subsequent centrifugation can separate more nomilin inclusion complexes, thereby effectively improving the content and transfer rate of nomilin in the bergamot extract. The content of nomilin in the obtained bergamot extract can be as high as 8% or more, and the yield can be as high as 60% or more. Compared with the existing process for extracting nomilin from plants, the content and yield of nomilin are higher.

[0164] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technician familiar with the field can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. A method for preparing a bergamot extract, characterized in that: include: After the bergamot fruit is crushed, it is subjected to wet granulation pre-treatment to form bergamot granules; Extracting the bergamot particles with supercritical carbon dioxide to obtain an extract; The extract is subjected to inclusion treatment by using cyclodextrin to form an inclusion liquid; The inclusion solution is concentrated under reduced pressure, cooled and allowed to stand, and centrifuged. After centrifugation, the precipitate is taken and dried to obtain the bergamot extract.

2. The method for preparing a bergamot extract according to claim 1, characterized in that: The steps of pre-treatment of wet granulation include: The crushed bergamot fruit is used as a powder material raw material, and an ethanol solution is used as a binder to prepare a soft material, and then the soft material is made into wet granules, and the wet granules are low-temperature dried and granulated to form the bergamot granules.

3. The method for preparing a bergamot extract according to claim 2, characterized in that: The ethanol solution is 50% to 95% ethanol, and the weight ratio of the powder material to the adhesive is 1:0.5 to 2.

4. The method for preparing a bergamot extract according to claim 1, characterized in that: The particle size of the bergamot particles is 30 meshes to 80 meshes.

5. The method for preparing a bergamot extract according to claim 1, characterized in that: The step of using cyclodextrin to carry out inclusion treatment on the extract comprises: dissolving cyclodextrin in water to prepare a saturated cyclodextrin aqueous solution; A saturated cyclodextrin aqueous solution is added to the extract while stirring, followed by high-speed shearing with a shearing machine or a colloid mill to form the inclusion liquid.

6. The method for preparing a bergamot extract according to claim 1, characterized in that: The cyclodextrin is one or more combinations of α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin.

7. The method for preparing a bergamot extract according to claim 1, characterized in that: After the reduced pressure concentration, high-speed shearing is performed using a shearing machine or a colloid mill, followed by cooling and standing; In the step of reducing pressure and concentrating, the concentration temperature is controlled below 55° C., and the concentrated solution is concentrated to a solid content mass fraction greater than 10%.

8. The method for preparing a bergamot extract according to claim 1, characterized in that: In the cooling and standing step, the cooling and standing time is more than 8 hours, and the cooling and standing temperature is 0°C to 8°C.

9. The method for preparing a bergamot extract according to claim 1, characterized in that: In the step of extracting the bergamot particles with supercritical carbon dioxide, the extraction time is 1h to 3h, the extraction temperature is 40°C to 50°C, the extraction pressure is 20MPa to 30MPa, the entrainer is ethanol, and the amount of the entrainer is 2 to 3 times the weight of the bergamot particles.

10. A bergamot extract, characterized in that The bergamot extract is prepared by the preparation method according to any one of claims 1 to 9.

Images