Tobacco root and stem extract, preparation method and use

Abstract

A preparation method for a tobacco root and stem extract, comprising the following steps: S1, cleaning tobacco roots and stems, removing impurities and then drying same; S2, crushing the dried tobacco roots and stems and sieving same to obtain tobacco root and stem powder; S3, weighing the tobacco root and stem powder, uniformly mixing same with a solvent, and subjecting same to ultrasonic cell disruption so as to obtain a solid-liquid mixed sample; and S4, subjecting the solid-liquid mixed sample to supercritical CO2 extraction, and, after the extraction is finished, mixing extract solutions obtained from a first-stage separation vessel and a second-stage separation vessel, so as to obtain a tobacco root and stem crude extract. The preparation method for obtaining the tobacco root and stem crude extract achieves higher yield compared with traditional preparation methods, and achieves higher purity compared with existing supercritical CO2 extraction methods. In addition, also provided is a method for further purifying the tobacco root and stem crude extract, which can obtain nicotine having higher purity.

Description

Tobacco rhizome extract, preparation method and application Technical Field

[0001] The invention belongs to the technical field of tobacco extracts, and in particular relates to a tobacco rhizome extract, a preparation method and an application thereof. Background Art

[0002] As we all know, nicotine is the main substance in tobacco that meets the physiological needs of consumers. At this stage, the main method is to add flavoring substances and nicotine to e-cigarette oil to simulate traditional cigarettes. While providing similar aroma and smoking experience, it avoids harmful substances produced by heating and combustion, such as carbon monoxide, tar, phenol, etc., and reduces the potential harm that e-cigarettes may cause to consumers and the public environment.

[0003] In recent years, with the steady development of the market for electronic heated cigarette atomizers, the demand for nicotine, or pure nicotine, has gradually increased worldwide.

[0004] The traditional industrial raw material for nicotine extraction is tobacco leaves, which are also the main raw material of traditional cigarettes. Generally, nicotine is extracted from ordinary tobacco, with a yield of between 2.5-3%.

[0005] Tobacco rhizomes are the discarded parts of tobacco plants and are generally considered tobacco waste. Currently, the disposal of tobacco rhizomes mainly involves burning and landfilling.

[0006] First, tobacco rhizomes can be processed by burning. This method converts tobacco rhizomes into heat energy and uses the organic matter in the rhizomes as fuel. However, burning produces a large amount of smoke and harmful gases, polluting the environment and wasting energy.

[0007] Secondly, tobacco rhizomes can also be disposed of through landfill. This method involves burying the rhizomes in the soil to allow them to gradually decompose. However, landfilling takes up land resources, and the decomposition process of tobacco rhizomes produces greenhouse gases such as methane, which has an impact on the atmospheric environment.

[0008] In summary, the current tobacco rhizomes not only fail to provide commercial value, but also require additional manpower and material costs for waste disposal, and the waste disposal process will still have an impact on the environment.

[0009] However, tobacco rhizomes contain a certain amount of nicotine. Extracting nicotine from tobacco rhizomes can not only reduce waste disposal costs, but also increase the comprehensive utilization rate of tobacco.

[0010] However, at present, there are very few means to utilize the tobacco rhizomes in tobacco waste, and the industry lacks relevant technologies to obtain nicotine with a high yield and utilization value from tobacco rhizomes.

[0011] Physical and Chemical Testing - Chemical Section, 2023, 59(05): 547-549, Ultrasonic-assisted extraction-high performance liquid chromatography was used to determine the content of nicotine in tobacco waste. The traditional method of extracting nicotine from tobacco leaves was disclosed. The nicotine extraction from tobacco rhizomes usually has a yield of about 0.1%.

[0012] Patent CN110437203A discloses a combined process of large-scale continuous countercurrent extraction and supercritical carbon dioxide extraction of nicotine, specifically discloses a combined process of large-scale continuous countercurrent extraction and supercritical carbon dioxide extraction of nicotine.

[0013] Patent CN112321564A discloses a process for extracting nicotine from waste tobacco leaves. Using supercritical CO₂ as the extraction agent, supercritical CO₂ is applied to the dried, pulverized, granulated, and dried waste tobacco pellets to produce a nicotine extract. This process simplifies the nicotine extraction process, requiring only drying, pulverization, and granulation, making it suitable for industrial, large-scale production.

[0014] The purity of the nicotine extract obtained in the specific embodiments of the above two patents is 40-55%.

[0015] It can be seen that the current technology for extracting nicotine from tobacco rhizomes is developing slowly, and nicotine from tobacco rhizomes cannot be extracted well, the yield is low, and the purity of the extracted nicotine is also relatively low.

[0016] In summary, how to provide a method for extracting nicotine from tobacco rhizomes is a technical problem that urgently needs to be solved.

[0017] Summary of the Invention

[0018] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a tobacco rhizome extract, a preparation method and an application thereof, comprising the following steps:

[0019] S1: Clean the tobacco rhizomes, remove impurities and then dry them;

[0020] S2 crushes and sieves the dried tobacco rhizomes to obtain tobacco rhizome powder;

[0021] S3: Weigh tobacco rhizome powder, mix it with solvent, and then disrupt the cells by ultrasonication to obtain a solid-liquid mixed sample;

[0022] S4 extracts the solid-liquid mixed sample through supercritical CO2. After the extraction is completed, the extract obtained from the first separation kettle is mixed with the extract obtained from the second separation kettle to obtain a crude tobacco rhizome extract.

[0023] The present invention can extract nicotine from tobacco rhizomes to obtain a crude tobacco rhizome extract.

[0024] The present invention provides a method for extracting nicotine from tobacco rhizomes, comprising the following steps:

[0025] S1: Clean the tobacco rhizomes, remove impurities and then dry them;

[0026] S2 crushes and sieves the dried tobacco rhizomes to obtain tobacco rhizome powder;

[0027] S3: Weigh tobacco rhizome powder, mix it with solvent, and then disrupt the cells by ultrasonication to obtain a solid-liquid mixed sample;

[0028] S4 extracts the solid-liquid mixed sample through supercritical CO2. After the extraction is completed, the extract obtained from the first separation kettle is mixed with the extract obtained from the second separation kettle to obtain a crude tobacco rhizome extract.

[0029] Furthermore, the solvent in S3 is ethanol, and the volume-to-mass ratio of the ethanol is between 1:1 and 1:5.

[0030] Furthermore, S3 ultrasonic cell disruption includes adding the sample to an ultrasonic cell disruption instrument and disrupting the sample at a frequency of 20-40 KHz for 5-10 minutes.

[0031] Furthermore, the extraction pressure during the supercritical CO2 extraction process is 16-25 MPa, the extraction temperature is 45-65°C, and the separation pressure is 4.5-6 MPa.

[0032] The present invention provides a tobacco rhizome crude extract, which is prepared by any one of the methods described above.

[0033] The present invention provides a method for purifying the above-mentioned crude tobacco rhizome extract, comprising the following steps: subjecting the crude tobacco rhizome extract to rotary evaporation and concentration to obtain a paste sample, and then subjecting the sample to purification to obtain a purified nicotine product.

[0034] Furthermore, the rotary evaporation concentration treatment of the tobacco rhizome crude extract includes:

[0035] The crude extract of tobacco rhizome is concentrated by rotary evaporation at 30-60°C until the solution turns dark brown, silica gel powder is added, and the mixture is rotary evaporated to dryness to obtain a mixed dry sample of nicotine extract and silica gel; the mixture is then infiltrated with triethylamine to obtain a paste sample.

[0036] Furthermore, the volume ratio of the triethylamine to the mixed dry sample is 1:5-1:10.

[0037] Furthermore, the purification process described in S5 includes purification process using column chromatography, comprising the following steps:

[0038] The column is packed with silica gel, the paste sample is added, and the mixture is eluted to obtain the purified nicotine extract;

[0039] The mixed liquid includes a methanol-dichloromethane mixed liquid, wherein the volume ratio of methanol to dichloromethane is 1:10-1:30.

[0040] The present invention provides nicotine, which is prepared by any one of the above methods for purifying a crude tobacco rhizome extract.

[0041] The present invention also provides a tobacco product containing the nicotine as described above.

[0042] Due to the adoption of the above technical solution, the present invention has the following advantages and positive effects compared with the prior art, as an example:

[0043] The crude tobacco rhizome extract prepared by the method provided by the present invention has a higher yield than that of traditional preparation methods, and has a higher purity than that of existing supercritical CO2 extraction methods.

[0044] At the same time, the present invention also provides a method for further purifying the obtained tobacco rhizome crude extract, which can obtain nicotine with higher purity.

[0045] The present invention helps to improve the utilization efficiency of tobacco resources, reduce the demand for traditional tobacco leaves, and open up new ways to utilize tobacco rhizomes. At the same time, it eliminates the need for extra manpower and material costs to treat tobacco rhizomes, helps to reduce the generation of tobacco waste and lower the impact on the environment.

[0046] In addition, the experimental method of the present invention can be expanded to a ton-level factory design, with high system efficiency, simple steps, and yield controlled within a range with corresponding commercial value, which is convenient for wide promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] FIG1 is a GC-MS diagram of the crude extract sample in Example 1 provided by the present invention

[0048] FIG2 is a GC-MS diagram of the crude extract sample in Example 2 provided by the present invention

[0049] FIG3 is a GC-MS diagram of the crude extract sample in Example 3 provided by the present invention

[0050] FIG4 is a GC-MS graph of pure nicotine in Example 4 provided by the present invention [0050.1] [Corrected 14.10.2024 according to Rule 91] Figure 5 shows the purity of the sample before purification in Example 4 provided by the present invention. DETAILED DESCRIPTION

[0051] The technical solutions disclosed in the present invention are described in detail below with reference to specific embodiments.

[0052] Techniques and methods known to those of ordinary skill in the relevant art may not be discussed in detail, but where appropriate, such techniques and methods should be considered part of the specification. In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0053] The present invention provides a method for extracting nicotine from tobacco rhizomes, comprising the following steps:

[0054] S1: Clean the tobacco rhizomes, remove impurities and then dry them.

[0055] Specifically, deionized water is usually used to clean the tobacco rhizomes, and after cleaning, the tobacco rhizomes are placed in an oven and dried at 80° C. to 100° C. for 8 to 12 hours.

[0056] S2 crushes and sieves the dried tobacco rhizomes to obtain tobacco rhizome powder.

[0057] Specifically, the dried tobacco rhizomes after drying are pulverized until they can pass through a 60-mesh sieve.

[0058] S3: Weigh tobacco rhizome powder, mix it evenly with the solvent, and then disrupt the cells by ultrasonication to obtain a solid-liquid mixed sample.

[0059] The solvent in S3 is ethanol, and the volume-to-mass ratio of ethanol is between 1:1 and 1:5, that is, the volume fraction is 60-100%.

[0060] The tobacco rhizome powder is evenly mixed with a solvent to obtain a tobacco rhizome powder extract or solution.

[0061] S3 ultrasonic cell disruption includes adding the sample to an ultrasonic cell disruption instrument and disrupting the sample at a frequency of 20-40 KHz for 5-10 minutes.

[0062] At this point, the pre-processing process is completed.

[0063] S4 extracts the solid-liquid mixed sample through supercritical CO2. After the extraction is completed, the extract obtained from the first separation kettle is mixed with the extract obtained from the second separation kettle to obtain a crude tobacco rhizome extract.

[0064] The instrument parameters in the supercritical CO2 extraction process are generally set as follows:

[0065] The extraction pressure is 25Mpa, the extraction temperature is 45-65℃, the separation pressure is 4.5-6Mpa, and the CO2 cylinder valve frequency is 16-32KHz.

[0066] S5: The crude tobacco rhizome extract is concentrated by rotary evaporation to obtain a paste sample, which is then purified to obtain a purified nicotine product.

[0067] Concentration is usually performed by rotary evaporation, which generally includes the following steps:

[0068] The crude extract of tobacco rhizome is concentrated by rotary evaporation at 30-60°C until the solution turns dark brown, silica gel powder is added, and the mixture is rotary evaporated to dryness to obtain a mixed dry sample of nicotine extract and silica gel; the mixture is then infiltrated with triethylamine to obtain a paste sample.

[0069] Preferably, the volume ratio of the added triethylamine to the mixed dry sample is 1:1-1:3.

[0070] The purification process includes using column chromatography to perform purification, including the following steps: packing a column with silica gel as filler, adding a paste sample, and eluting with a mixed solution to obtain a purified nicotine extract.

[0071] The mixed liquid here can be a methanol-dichloromethane mixed liquid, wherein the volume ratio of methanol to dichloromethane is 1:10-1:30.

[0072] The present invention provides a tobacco rhizome crude extract, which is prepared by any one of the methods described above.

[0073] The present invention provides a method for purifying the above-mentioned crude tobacco rhizome extract, comprising the following steps: subjecting the crude tobacco rhizome extract to rotary evaporation and concentration to obtain a paste sample, and then subjecting the sample to purification to obtain purified nicotine.

[0074] The present invention also provides nicotine, which is prepared by any one of the above methods for purifying a crude tobacco rhizome extract.

[0075] The present invention also provides a tobacco product containing the above-mentioned pure nicotine.

[0076] Example 1:

[0077] The tobacco rhizomes were washed with deionized water to remove dirt and residue, and then placed in a 100°C oven for drying for about 8 hours. The dried tobacco rhizomes were crushed until they could pass through a 60-mesh sieve.

[0078] Weigh 60g of the obtained tobacco rhizome powder and mix it with anhydrous ethanol at a volume-to-mass ratio of 1:1 to obtain a tobacco rhizome powder extract or solution. Add the sample to an ultrasonic cell disruptor and disrupt it at a frequency of 20 kHz for 10 minutes.

[0079] The solid-liquid mixed sample after pretreatment was added to the supercritical CO2 extraction instrument. The instrument parameters were adjusted to set the extraction pressure to 25 MPa, the temperature to 65°C, the separation pressure to 5.6 MPa, the CO2 cylinder valve frequency to 16 KHz, and the entire extraction process time to 120 minutes.

[0080] After the extraction is completed, the fractions in the first separation kettle and the second separation kettle are collected and mixed to obtain about 57 ml of an ethanol solution of tobacco rhizome extract.

[0081] The crude extract was quantitatively analyzed by GC-MS. The results are shown in FIG1 . The nicotine concentration was approximately 2.31 mg / ml, and the yield was calculated to be approximately 0.22%.

[0082] Example 2:

[0083] The tobacco rhizomes were washed with deionized water to remove dirt and residue, and then placed in a 100°C oven for drying for about 8 hours. The dried tobacco rhizomes were crushed until they could pass through a 60-mesh sieve.

[0084] Weigh 25g of the obtained tobacco rhizome powder and mix it with anhydrous ethanol at a volume-to-mass ratio of 1:5 to obtain a tobacco rhizome powder extract or solution. Add the sample to an ultrasonic cell disruptor and disrupt it at a frequency of 20 kHz for 10 minutes.

[0085] The solid-liquid mixed sample after pretreatment was added to the supercritical CO2 extraction instrument. The instrument parameters were adjusted to set the extraction pressure to 25 MPa, the temperature to 65°C, the separation pressure to 5.6 MPa, the CO2 cylinder valve frequency to 16 KHz, and the entire extraction process time to 120 minutes.

[0086] After the extraction is completed, the fractions in the first separation kettle and the second separation kettle are collected and mixed to obtain about 135 ml of an ethanol solution of tobacco rhizome extract.

[0087] The obtained crude extract was quantitatively analyzed by GC-MS instrument. The results are shown in FIG2 . The nicotine concentration was about 0.71 mg / ml, and the yield was calculated to be about 0.38%.

[0088] Example 3:

[0089] The tobacco rhizomes were washed with deionized water to remove dirt and residue, and then placed in a 100°C oven for drying for about 8 hours. The dried tobacco rhizomes were crushed until they could pass through a 60-mesh sieve.

[0090] Weigh 25g of the obtained tobacco rhizome powder and mix it with 90% ethanol at a volume-to-mass ratio of 1:5 to obtain a tobacco rhizome powder extract or solution. Add the sample to an ultrasonic cell disruptor and disrupt it at a frequency of 20 kHz for 10 minutes.

[0091] The solid-liquid mixed sample after pretreatment was added to the supercritical CO2 extraction instrument. The instrument parameters were adjusted to set the extraction pressure to 25 MPa, the temperature to 65°C, the separation pressure to 5.6 MPa, the CO2 cylinder valve frequency to 16 KHz, and the entire extraction process time to 120 minutes.

[0092] After the extraction is completed, the fractions in the first separation kettle and the second separation kettle are collected and mixed to obtain about 118 ml of an ethanol solution of tobacco rhizome extract.

[0093] The obtained crude extract was quantitatively analyzed by GC-MS instrument. The results are shown in FIG3 . The nicotine concentration was about 1.035 mg / ml, and the yield was calculated to be about 0.49%.

[0094] The above experimental results show that the crude tobacco rhizome extract obtained by the method provided by the present invention has a higher yield than the traditional preparation method. After optimization by the present invention, the maximum yield can reach 0.49%, and the extract has a high purity, with an average purity of about 61%.

[0095] At the same time, the present invention also provides a method for further purifying the obtained tobacco rhizome crude extract.

[0096] Example 4:

[0097] The crude tobacco rhizome extract was concentrated by rotary evaporation at 60°C for approximately 15-30 minutes until the solution turned dark brown. Silica gel powder was then added and rotary evaporation continued until dryness was achieved, yielding a dry mixture of nicotine extract and silica gel. This mixture was then infiltrated with 2 volumes of triethylamine to obtain a paste. A column was then packed with 200-400 mesh silica gel, and the extract was added. The column was then eluted with a mixture of methanol and dichloromethane at a volume ratio of 1:10. The eluate was collected and subjected to rotary evaporation with suction to obtain purified nicotine with a purity of approximately 98%.

[0098] The obtained pure nicotine was quantitatively analyzed by GC-MS instrument, and the results are shown in FIG4 .

[0099] [Corrected 14.10.2024 according to Rule 91] The purity of the sample before purification in Example 4 of the present invention is shown in Figure 5.

[0100] [Corrected 14.10.2024 in accordance with Article 91]

[0101] [Corrected 14.10.2024 according to Rule 91] Table 1 shows the purity of the sample after purification in Example 4 of the present invention

[0102] [Corrected 14.10.2024 according to Rule 91] Table 1 Purity of the purified samples

[0103] The above experimental results indicate that nicotine with a relatively high purity can be obtained by further purifying the crude tobacco rhizome extract obtained by the method provided by the present invention.

[0104] Within the scope of protection intended by the present disclosure, terms such as "including" and "comprising" should be interpreted as inclusive or open-ended rather than exclusive or closed by default, unless expressly defined to the contrary. All technical, scientific, or other terms have the meanings understood by those skilled in the art unless expressly defined to the contrary. Common terms found in dictionaries should not be interpreted in an overly idealistic or unrealistic manner in the context of relevant technical documents, unless expressly defined to that extent by the present disclosure.

[0105] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims, not the foregoing description, and all variations within the meaning and range of equivalents of the claims are intended to be included therein. Any reference sign in a claim should not be construed as limiting the claim to which it relates.

[0106] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A method for extracting nicotine from tobacco rhizomes, characterized in that, It includes the following steps: S1: Clean the tobacco rhizomes, remove impurities, and then dry them. S2: Crush the dried tobacco rhizomes and sieve them to obtain tobacco rhizome powder. S3: Weigh the tobacco rhizome powder, mix it evenly with a solvent, and then obtain a solid-liquid mixed sample through ultrasonic cell disruption. S4: Subject the solid-liquid mixed sample to supercritical CO2 extraction. After the extraction is completed, mix the extraction liquid phases obtained from the first-stage separation kettle and the second-stage separation kettle to obtain a crude tobacco rhizome extract.

2. The method for extracting nicotine from tobacco rhizomes according to claim 1, characterized in that: The solvent in S3 is ethanol, and the volume-mass ratio of the ethanol is between 1:1 and 1:

5. The ultrasonic cell disruption in S3 includes adding the sample to an ultrasonic cell disruption instrument and disrupting it at a frequency of 20 - 40 KHz for 5 - 10 minutes.

3. The method for extracting nicotine from tobacco rhizomes according to claim 1, characterized in that: In S4, the extraction pressure during the supercritical CO2 extraction process is 16 - 25 Mpa, the extraction temperature is 45 - 65 °C, and the separation pressure is 4.5 - 6 Mpa.

4. A crude extract of tobacco rhizome, characterized in that, It is prepared by the method described in any one of claims 1 to 3.

5. A method for purifying the crude extract of tobacco rhizome as described in claim 4, characterized in that, It includes the following steps: Rotate and evaporate and concentrate the crude tobacco rhizome extract to obtain a paste-like sample, and then through purification treatment, obtain purified nicotine pure product.

6. The method for extracting nicotine from tobacco rhizomes according to claim 5, characterized in that: The rotation and evaporation and concentration treatment of the crude tobacco rhizome extract includes Rotating and evaporating and concentrating the crude tobacco rhizome extract at 30 °C - 60 °C until the solution turns dark brown, adding silica powder, and rotating and evaporating to dryness to obtain a mixed dry sample of nicotine extract and silica; then infiltrating with triethylamine to obtain a paste-like sample.

7. The method for extracting nicotine from tobacco rhizomes according to claim 6, characterized in that: The volume ratio of the triethylamine to the mixed dry sample is 1:5 - 1:

10.

8. The method according to claim 5, characterized in that: The purification treatment in S5 includes using column chromatography for purification treatment, including the following steps Pack a column with silica as the packing material, add the paste-like sample, and elute with a mixed solution to obtain a purified nicotine extract. The mixed solution includes a methanol-dichloromethane mixed solution, wherein the volume ratio of methanol to dichloromethane is 1:10 - 1:

30.

9. A nicotine, characterized in that, It is prepared by the method for purifying the crude tobacco rhizome extract described in any one of claims 5 to 8.

10. A tobacco product, characterized in that, It contains nicotine as described in claim 9.

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