Method for extracting and purifying solanesol in waste tobacco leaves

Abstract

The invention belongs to a process for extracting, separating and purifying fat-soluble chemical components, and particularly discloses a method for extracting and purifying solanesol in waste and inferior tobacco leaves. According to the method, the solanesol in the waste and inferior tobacco leaves is subjected to green extraction by adopting a lauryl sodium sulfate aqueous solution, the solanesol in an extracting solution is subjected to directional enrichment by utilizing the charge characteristic of chitosan, the solanesol in an enriched product is extracted by ethyl acetate, the combined solanesol is saponified by sodium hydroxide, and recrystallization and anti-solvent crystallization purification are carried out. The high-purity solanesol is prepared. The extraction solvent used in the method is green and safe, the energy consumption of the extraction and purification process is low, process parameters are easy to control, the production period is shortened, and the method is suitable for industrial application and market promotion.

Description

Technical Field

[0001] The invention belongs to a process for extracting, separating and purifying fat-soluble chemical components, and specifically discloses a method for extracting and purifying solanesol from waste and inferior tobacco leaves. Background Art

[0002] Tobacco is one of the most widely cultivated plants. From tobacco cultivation to the production of tobacco products, it includes a large amount of low-grade tobacco leaves. If not utilized, it will cause serious damage to the environment. Tobacco is the richest source of solanesol, with the solanesol content reaching about 4% of the dry weight of tobacco. Therefore, waste tobacco leaves can be used as a raw material for extracting solanesol, which not only brings economic benefits but also reduces environmental pressure. Solanesol is a polyterpenoid compound contained in tobacco, an unsaturated polyisoprene alcohol. It is a white solid powder with weak polarity and high medicinal value. It has antibacterial, anti-inflammatory, anti-cancer and hemostatic effects. Solanesol is also a synthetic coenzyme Q. 10 , anti-ulcer drugs, vitamin K2 and other pharmaceutical products.

[0003] Currently, the extraction and purification of solanesol often involves the use of hazardous solvents such as petroleum ether, acetone, and n-hexane, which are explosive and unsuitable for large-scale industrial production. Therefore, establishing an efficient and low-risk solanesol extraction and purification process could further promote the development of solanesol's deep processing industry, provide favorable technical support for the high-value utilization of waste tobacco, and be key to the commercialization of solanesol.

[0004] Chinese patents CN113264812A and CN112225641A use n-hexane and petroleum ether as extraction or purification solvents. The solvents are extremely flammable and explosive when exposed to open flames and high heat, and are therefore dangerous to use on a large scale in industry. Summary of the Invention

[0005] In response to the above invention, the present invention aims to provide a new green extraction and purification process for solanesol. The present invention proposes a series of new extraction and purification steps including micellar extraction, chitosan flocculation separation, ethyl acetate extraction, sodium hydroxide saponification, recrystallization and anti-solvent crystallization purification, which has low risk and high extraction rate and purity of solanesol.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The process for extracting and purifying solanesol from waste and inferior tobacco leaves of the present invention comprises the following steps:

[0008] (1) crushing waste tobacco leaves and drying them in the sun to obtain tobacco leaf powder;

[0009] (2) Mix the tobacco leaf powder with sodium dodecyl sulfate micelles at a certain solid-liquid ratio to obtain a mixture. Extract the mixture to obtain an extract. Centrifuge the extract and take the supernatant to obtain a solanesol extract.

[0010] (3) Add an aqueous solution of chitosan acetate to the solanesol extract, stir and mix to obtain a mixed solution. Centrifuge the mixed solution to obtain a solanesol enrichment, and dry the enrichment.

[0011] (4) Add ethyl acetate to the dried enrichment for extraction. Centrifuge the extraction solution and take the supernatant, and spin-dry the ethyl acetate to obtain an ethyl acetate extract.

[0012] (5) Add a sodium hydroxide ethanol solution to the ethyl acetate extract for saponification. Neutralize the saponified solution with HCl to adjust the pH value to about 7. Centrifuge the solution to obtain the supernatant, and spin-dry it to obtain a solanesol extract paste.

[0013] (6) Dissolve the solanesol extract paste in a methanol solution, perform low-temperature recrystallization, centrifuge and take out the crystal to obtain crude solanesol.

[0014] (7) Dissolve the crude solanesol in an ethanol solution, add deionized water for anti-solvent crystallization, centrifuge the solution to obtain pure solanesol.

[0015] Preferably, in step (2), the mass percentage content of sodium dodecyl sulfate micelles is 0.5-9 wt%, and the mass ratio of the tobacco leaf powder to the volume of sodium dodecyl sulfate micelles is 1 g:10-35 ml; the extraction includes one or more of heating reflux extraction, microwave extraction, ultrasonic extraction or ultra-high pressure extraction; the number of heating reflux extractions is 1-3 times, and the extraction time for each time is 30-60 min; the number of microwave extractions is 1-3 times, and the time for each time is 5-30 min; the number of ultrasonic extractions is 1-3 times, and the time for each time is 30-60 min; the number of ultra-high pressure extractions is 1-3 times, the extraction time for each time is 0.5-3 min, and the extraction pressure for each time is 50-300 MPa.

[0016] Preferably, in step (3), the mass percentage content of chitosan in the aqueous solution of chitosan acetate is 0.5-3 wt%; the addition amount of the aqueous solution of chitosan acetate is 50-100% of the volume of the solanesol extract, and the stirring and mixing time of the aqueous solution of chitosan acetate and the solanesol extract is 1-3 h.

[0017] Preferably, in step (4), the mass ratio of the dried enrichment to the volume of the ethyl acetate extraction solution is 1 g:4-9 ml, the extraction temperature is 10-35 °C, and the extraction time is 1-3 h.

[0018] Preferably, in the step (5), the concentration of the sodium hydroxide ethanol solution is 0.02 - 0.2 mol / L, the saponification time is 1 - 4 h, and the saponification temperature is 45 - 65 °C.

[0019] Preferably, in the step (6), the concentration of the solanesol extract is 50 - 250 mg / ml, the recrystallization temperature is -20 - 20 °C, and the crystallization time is 4 - 20 h.

[0020] Preferably, in the step (7), the concentration of the crude solanesol is 50 - 250 mg / ml, and the volume ratio of the solanesol solution for anti-solvent crystallization to deionized water is 1:10 - 1:50.

[0021] Preferably, in the steps (2) - (7), the centrifugation speed is 6000 - 10000 r / min, and the centrifugation time is 6 - 10 min.

[0022] The present invention provides a new process for extracting and purifying solanesol from waste and inferior tobacco leaves, comprising the following steps: crushing waste and inferior tobacco leaves, drying in the sun to obtain tobacco leaf powder; mixing the tobacco leaf powder with sodium dodecyl sulfate micelles at a certain solid-liquid ratio to obtain a mixture, extracting the mixture to obtain an extract, centrifuging the extract, taking the supernatant to obtain a solanesol extract. Adding an aqueous chitosan acetate solution to the solanesol extract, stirring and mixing to obtain a mixed solution, centrifuging the mixed solution to obtain a solanesol enrichment, and drying the enrichment. Adding ethyl acetate to the dried enrichment for extraction, centrifuging the extract, taking the supernatant, and rotary-evaporating the ethyl acetate to obtain an ethyl acetate extract. Adding a sodium hydroxide ethanol solution to the ethyl acetate extract for saponification, neutralizing the saponified solution with HCl to adjust the pH value to about 7, centrifuging the solution to obtain the supernatant, and rotary-evaporating to obtain a solanesol extract. Dissolving the solanesol extract in a methanol solution, performing low-temperature recrystallization, centrifuging to take out the crystalline substance to obtain crude solanesol. Dissolving the crude solanesol in an ethanol solution, adding deionized water for anti-solvent crystallization, centrifuging the solution to obtain pure solanesol.

[0023] An aqueous solution of sodium dodecyl sulfate is used to replace the organic solvents used in the extraction of solanesol in traditional industries. The solvent is green and safe, and has no disadvantages such as flammability, explosiveness, and high harm to the human body compared with traditional extraction solvents. At the same time, the production cost is reduced, and the maximum benefit of tobacco is achieved. Sodium dodecyl sulfate and solanesol generate hydrogen bond forces during the extraction process, which can wrap solanesol in the hydrophobic core to form a supramolecular system. According to the charge characteristics of the supramolecular interface formed by sodium dodecyl sulfate and solanesol, the charge adsorption effect of chitosan on sodium dodecyl sulfate is used for the enrichment and separation of solanesol. Ethyl acetate is used to extract the solanesol component in the enrichment, sodium hydroxide is used to saponify the bound solanesol, and anti-solvent crystallization and recrystallization are used to purify the solanesol extract. The solvent used in the present invention has low risk, the method is simple to operate, and the energy consumption and equipment requirements are low. The maximum extraction rate of solanesol by the extraction method is 97.76%, and the maximum purity is 92.54%. Description of the Drawings

[0024] Figure 1 It is the high performance liquid chromatography diagram of the extraction process; Figure 1 Among them, (a) is the liquid chromatography diagram of the solanesol standard product, Figure 1 Among them, (b) is the liquid chromatography diagram of the solanesol extract.

[0025] Figure 2 It is the high performance liquid chromatography diagram of the purification process; Figure 2 Among them, (a) is the liquid chromatography diagram of the solanesol standard product, Figure 2 Among them, (b) is the liquid chromatography diagram of the extract, Figure 2 Among them, (c) is the liquid phase diagram of the crude solanesol after recrystallization, Figure 2 Among them, (d) is the liquid chromatography diagram of the pure solanesol after anti-solvent crystallization. Detailed Embodiment

[0026] The present invention provides a new process for the sustainable extraction and purification of solanesol, including the following steps:

[0027] (1) Crush and dry the waste and inferior tobacco leaves to obtain tobacco leaf powder;

[0028] (2) Mix the tobacco leaf powder and sodium dodecyl sulfate micelles according to a certain solid-liquid ratio to obtain a mixture. The mixture is extracted to obtain an extract. The extract is centrifuged, and the supernatant is taken to obtain a solanesol extract;

[0029] (3) Add an aqueous solution of chitosan acetate to the solanesol extract, stir and mix to obtain a mixed solution. The mixed solution is centrifuged to obtain a solanesol enrichment, and the enrichment is dried;

[0030] (4) Add ethyl acetate to the dried enrichment for extraction. The extraction solution is centrifuged, and the supernatant is taken and the ethyl acetate is evaporated to dryness to obtain an ethyl acetate extract;

[0031] (5) Add sodium hydroxide ethanol solution to the ethyl acetate extract for saponification. After saponification, neutralize the solution with HCl, adjust the pH value to about 7, centrifuge the solution to obtain the supernatant, and spin-dry to obtain solanesol extract.

[0032] (6) Dissolve the solanesol extract in methanol solution, perform low-temperature recrystallization, centrifuge to remove the crystalline substance to obtain crude solanesol.

[0033] (7) Dissolve the crude solanesol in ethanol solution, add deionized water for anti-solvent crystallization, centrifuge the solution to obtain pure solanesol.

[0034] In the present invention, waste and inferior tobacco leaves are crushed and dried to obtain tobacco leaf powder.

[0035] Mix the tobacco leaf powder obtained in step (1) with sodium dodecyl sulfate micelles at a certain solid-liquid ratio to obtain a mixture.

[0036] In the present invention, the micelle extraction solvent in step (2) is sodium dodecyl sulfate, and its concentration is preferably 5 - 90 mg / ml, more preferably 10 - 50 mg / ml. The present invention has no special limitation on the source of the sodium dodecyl sulfate solution, and it can be prepared by a preparation method well-known to those skilled in the art.

[0037] In the present invention, the dosage ratio of the tobacco leaf powder to the micelle extraction solvent is preferably 1 g:10 - 35 ml, more preferably 1 g:10 - 30 ml.

[0038] In the present invention, the extraction preferably includes one or more of heating reflux extraction, microwave extraction, ultrasonic extraction or ultra-high pressure extraction, more preferably ultra-high pressure extraction, heating reflux extraction or ultrasonic extraction. The number of times of heating reflux extraction is preferably 1 - 3 times, and the extraction time each time is preferably 30 - 60 min, more preferably 45 - 60 min. The number of times of microwave extraction is preferably 1 - 3 times, and the time each time is preferably 5 - 30 min, more preferably 20 - 30 min. The number of times of ultrasonic extraction is preferably 1 - 3 times, and the time each time is preferably 30 - 60 min, more preferably 45 - 60 min. The number of times of ultra-high pressure extraction is preferably 1 - 3 times, the extraction time each time is preferably 0.5 - 3 min, more preferably 2 - 3 min, and the extraction pressure each time is preferably 50 - 300 MPa, more preferably 200 - 300 MPa.

[0039] After the extraction is completed, centrifuge the mixture, take the supernatant to obtain the solanesol extract, and determine the content of solanesol in the extract.

[0040] In the present invention, the mass percentage of chitosan in the chitosan acetic acid aqueous solution in step (3) is preferably 0.5-3 wt%, more preferably 2-3 wt%; the addition amount of the chitosan acetic acid aqueous solution is preferably 50-100% of the volume of the solanesol extract, more preferably 75-100% of the volume of the solanesol extract; the stirring and mixing time of the chitosan acetic acid aqueous solution and the solanesol extract is preferably 1-3 h, more preferably 2-3 h.

[0041] A mixed solution is obtained, the mixed solution is centrifuged to obtain a solanesol concentrate, and the concentrate is dried.

[0042] In the present invention, the mass ratio of the dried concentrate in step (4) to the volume of the ethyl acetate extractant is preferably 1 g:4-9 ml, more preferably 1 g:7-9 ml; the extraction temperature is preferably 10-35 °C, more preferably 10-20 °C; the extraction time is preferably 1-3 h, more preferably 2-3 h.

[0043] Sodium hydroxide ethanol solution is added to the ethyl acetate extract in step (5) for saponification. The concentration of the sodium hydroxide ethanol solution is preferably 0.02-0.2 mol / L, more preferably 0.1-0.2 mol / L; the saponification time is preferably 1-4 h, more preferably 3-4 h; the saponification temperature is preferably 45-65 °C, more preferably 55-65 °C, to obtain a solanesol extract, and the purity of the solanesol extract is measured.

[0044] In the present invention, the solanesol extract in step (6) is dissolved in a methanol solution. The concentration of the solanesol extract is preferably 50-250 mg / ml, more preferably 150-250 mg / ml. It is recrystallized at low temperature. The temperature is preferably -20 °C to 20 °C, more preferably -20 °C to 4 °C; the time is preferably 4-20 h, more preferably 4-12 h. The crystals are taken out by centrifugation to obtain crude solanesol, and the purity of the crude solanesol is measured.

[0045] In the present invention, the crude solanesol obtained in step (7) is dissolved in ethanol and deionized water is added for anti-solvent crystallization. The concentration of the crude solanesol is preferably 50-250 mg / ml, more preferably 150-250 mg / ml. The volume ratio of ethanol to water is preferably in the range of 1:10-1:50, more preferably 1:30-1:50. The solution is centrifuged to obtain pure solanesol, and the purity of the pure solanesol is measured.

[0046] In the present invention, the centrifugation speed in steps (2)-(7) is preferably 6000-10000 r / min, more preferably 8000-10000 r / min; the centrifugation time is preferably 6-10 min, more preferably 8-10 min.

[0047] To further illustrate the present invention, the method for extracting solanesol from waste and substandard tobacco provided by the present invention will be described in detail below with reference to examples, but they should not be construed as limiting the scope of protection of the present invention.

[0048] Example 1:

[0049] The waste and substandard tobacco was pulverized and dried in the sun to obtain tobacco powder. 1 g of the tobacco powder was weighed and thoroughly mixed with 10 ml of a sodium dodecyl sulfate micelle solution with a concentration of 90 mg / ml to obtain a mixture. The mixture was placed in a high-pressure equipment for high-pressure extraction. The extraction was carried out once, with an extraction time of 3 min and an extraction pressure of 50 MPa. After the extraction, the mixture was centrifuged at a rotational speed of 6000 r / min for 10 min, and the supernatant was taken to obtain a solanesol extract. The content of solanesol in the extract was determined by high-performance liquid chromatography; through calculation, the extraction rate of solanesol was 77.76%. According to 50% of the volume of the solanesol extract, an aqueous solution of chitosan acetate with a mass percentage content of 3 wt% was added, and the stirring and mixing time of the aqueous solution of chitosan acetate and the solanesol extract was 1 h to obtain a mixed solution. The mixed solution was centrifuged to obtain a solanesol enrichment product, and the enrichment product was dried. The enrichment rate of solanesol was 97.95%. 1 g of the dried enrichment product was taken, 9 ml of ethyl acetate was added, the extraction temperature was 10 °C, and the extraction time was 3 h. The extract was centrifuged at a rotational speed of 10000 r / min for 6 min, and the supernatant was taken and the ethyl acetate was evaporated to dryness to obtain an ethyl acetate extract. 0.2 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract, and saponification was carried out at 45 °C for 1 h. After saponification, the solution was neutralized with HCl to adjust the pH value to about 7. The solution was centrifuged to obtain the supernatant, which was evaporated to dryness to obtain a solanesol extract paste, and the purity of the solanesol extract paste was 30.55%. 250 mg of the solanesol extract paste was dissolved in 1 ml of methanol solution, and recrystallization was carried out at 20 °C for 4 h. The crystalline product was centrifuged out to obtain a crude solanesol product, and the purity of the crude solanesol product was 57.56%. 250 mg of the crude solanesol product was dissolved in 1 ml of ethanol solution, 10 ml of deionized water was added for anti-solvent crystallization. The solution was centrifuged to obtain a pure solanesol product, and the purity of the pure solanesol product was 89.20%.

[0050] Example 2:

[0051] Waste tobacco is crushed and sun-dried to obtain tobacco powder. One gram of tobacco powder is weighed and thoroughly mixed with 35 ml of 5 mg / ml sodium lauryl sulfate micelles to obtain a mixture. The mixture is then placed in a heated reflux extraction apparatus for extraction, with one extraction cycle and a time of 60 minutes. After extraction, the mixture is centrifuged at 10,000 rpm for 9 minutes. The supernatant is collected to obtain a solanesol extract, which is then assayed for solanesol content by high-performance liquid chromatography. The solanesol extraction yield is calculated to be 80.22%. A chitosan acetate aqueous solution (0.5 wt% by weight) is added to 100% of the solanesol extract. The chitosan acetate aqueous solution and the solanesol extract are stirred for 3 hours to obtain a mixed solution, which is then centrifuged to obtain a solanesol concentrate. The concentrate is then dried, and the solanesol enrichment yield is 81.66%. 1 g of the dried concentrate was added to 4 ml of ethyl acetate. The extraction temperature was 35°C for 1 hour. The extract was centrifuged at 9000 r / min for 6 minutes. The supernatant was removed and the ethyl acetate was spin-dried to obtain an ethyl acetate extract. 0.02 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract and saponified at 65°C for 4 hours. The saponified solution was neutralized with HCl and the pH was adjusted to approximately 7. The solution was centrifuged to obtain the supernatant, which was spin-dried to obtain a solanesol extract with a purity of 25.72%. 50 mg of the solanesol extract was dissolved in 1 ml of methanol and recrystallized at -20°C for 20 hours. The crystals were removed by centrifugation to obtain crude solanesol with a purity of 67.56%. 50 mg of crude solanesol was dissolved in 1 ml of ethanol solution, and 50 ml of deionized water was added for anti-solvent crystallization. The solution was centrifuged to obtain pure solanesol. The purity of the pure solanesol was 90.48%.

[0052] Example 3:

[0053] Waste tobacco is crushed and sun-dried to obtain tobacco powder. One gram of tobacco powder is weighed and thoroughly mixed with 30 ml of 50 mg / ml sodium lauryl sulfate micelles to obtain a mixture. The mixture is then subjected to a microwave extraction device for extraction, with one extraction cycle and a time of 30 minutes. After extraction, the mixture is centrifuged at 8000 rpm for 7 minutes. The supernatant is collected to obtain a solanesol extract, which is then assayed for solanesol by high performance liquid chromatography. The solanesol extraction yield is calculated to be 88.99%. A 2 wt% aqueous chitosan acetate solution is added to 90% of the volume of the solanesol extract. The chitosan acetate solution and the solanesol extract are stirred for 2 hours to obtain a mixed solution, which is then centrifuged to obtain a solanesol concentrate. The concentrate is then dried, and the solanesol enrichment yield is 100%. 1 g of the dried concentrate was added to 7 ml of ethyl acetate. The extraction temperature was 35°C for 2 hours. The extract was centrifuged at 7000 rpm for 9 minutes. The supernatant was removed and the ethyl acetate was spin-dried to obtain an ethyl acetate extract. 0.1 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract and saponified at 55°C for 3 hours. The saponified solution was neutralized with HCl and the pH was adjusted to approximately 7. The solution was centrifuged to obtain the supernatant, which was spin-dried to obtain a solanesol extract with a purity of 29.66%. 200 mg of the solanesol extract was dissolved in 1 ml of methanol and recrystallized at -20°C for 16 hours. The crystals were removed by centrifugation to obtain crude solanesol with a purity of 68.23%. 200 mg of crude solanesol was dissolved in 1 ml of ethanol solution, 30 ml of deionized water was added, and anti-solvent crystallization was performed. The solution was centrifuged to obtain pure solanesol. The purity of the pure solanesol was 91.76%.

[0054] Embodiment 4:

[0055] Waste tobacco is crushed and sun-dried to obtain tobacco powder. One gram of tobacco powder is weighed and thoroughly mixed with 25 ml of 30 mg / ml sodium lauryl sulfate micelles to obtain a mixture. The mixture is then subjected to an ultrasonic extraction apparatus for extraction, with one extraction cycle lasting 60 minutes. After extraction, the mixture is centrifuged at 7000 rpm for 6 minutes. The supernatant is collected to obtain a solanesol extract, which is then assayed for solanesol content by high-performance liquid chromatography. The solanesol extraction yield is calculated to be 82.56%. A 2.5 wt% aqueous chitosan acetate solution is added to 80% of the solanesol extract by volume. The chitosan acetate solution and the solanesol extract are stirred for 1.5 hours to obtain a mixed solution, which is then centrifuged to obtain a solanesol concentrate. The concentrate is then dried, resulting in a solanesol enrichment yield of 95.02%. 1 g of the dried concentrate was added to 6 ml of ethyl acetate. The extraction temperature was 15°C for 1.5 hours. The extract was centrifuged at 10,000 rpm for 6 minutes. The supernatant was removed and the ethyl acetate was spin-dried to obtain an ethyl acetate extract. 0.15 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract and saponified at 50°C for 2 hours. The saponified solution was neutralized with HCl and the pH was adjusted to approximately 7. The solution was centrifuged to obtain the supernatant, which was spin-dried to obtain a solanesol extract with a purity of 27.88%. 100 mg of the solanesol extract was dissolved in 1 ml of methanol and recrystallized at 0°C for 20 hours. The crystals were removed by centrifugation to obtain crude solanesol with a purity of 54.21%. 150 mg of crude solanesol was dissolved in 1 ml of ethanol solution, 30 ml of deionized water was added, and anti-solvent crystallization was performed. The solution was centrifuged to obtain pure solanesol. The purity of the pure solanesol was 92.54%.

[0056] Example 5:

[0057] The waste and inferior tobacco is crushed and dried in the sun to obtain tobacco powder. Weigh 1 g of the tobacco powder and mix it thoroughly with 15 ml of a sodium dodecyl sulfate micelle solution with a concentration of 70 mg / ml to obtain a mixture. Place the mixture in a high-pressure equipment for ultra-high pressure extraction. The extraction is carried out 3 times, with each extraction time being 0.5 min and the extraction pressure being 300 MPa. After the extraction is completed, the mixture is centrifuged at a rotational speed of 6000 r / min for 10 min. Take the supernatant to obtain a solanesol extract, and determine the content of solanesol in the extract by high-performance liquid chromatography; through calculation, the extraction rate of solanesol is 97.76%. Add an aqueous solution of chitosan acetate with a mass percentage content of 1.5 wt% according to 60% of the volume of the solanesol extract. The stirring and mixing time of the aqueous solution of chitosan acetate and the solanesol extract is 2 h to obtain a mixed solution. The mixed solution is centrifuged to obtain a solanesol enrichment product, and the enrichment product is dried. The enrichment rate of solanesol is 93.37%. Take 1 g of the dried enrichment product, add 8 ml of ethyl acetate, the extraction temperature is 20 °C, the extraction time is 2 h, the extract is centrifuged at a rotational speed of 10000 r / min for 6 min, take the supernatant, and evaporate the ethyl acetate to dryness to obtain an ethyl acetate extract. Add a 0.2 mol / L sodium hydroxide ethanol solution to the ethyl acetate extract, saponify at 60 °C for 2 h, neutralize the saponified solution with HCl, adjust the pH value to about 7, centrifuge the solution, take the supernatant, evaporate to dryness to obtain a solanesol extract paste, and the purity of the solanesol extract paste is 27.52%. Dissolve 200 mg of the solanesol extract paste in 1 ml of methanol solution, recrystallize at -20 °C for 8 h, centrifuge to take out the crystalline substance to obtain a crude solanesol product, and the purity of the crude solanesol product is 60.17%. Dissolve 200 mg of the crude solanesol product in 1 ml of ethanol solution, add 20 ml of deionized water for antisolvent crystallization, centrifuge the solution to obtain a pure solanesol product, and the purity of the pure solanesol product is 92.22%.

[0058] Example 6:

[0059] Waste tobacco is crushed and sun-dried to obtain tobacco powder. One gram of tobacco powder is weighed and thoroughly mixed with 25 ml of 30 mg / ml sodium lauryl sulfate micelles to obtain a mixture. The mixture is then placed in a heated reflux extraction apparatus for extraction, with three extractions of 30 minutes each. After the extractions are completed, the mixture is centrifuged at 10,000 rpm for 9 minutes. The supernatant is collected to obtain a solanesol extract, which is then assayed for solanesol content by high-performance liquid chromatography. The solanesol extraction yield is calculated to be 93.54%. A 1 wt% aqueous chitosan acetate solution is added to 80% of the volume of the solanesol extract. The chitosan acetate solution and the solanesol extract are stirred for 3 hours to obtain a mixed solution, which is then centrifuged to obtain a solanesol concentrate. The concentrate is then dried, resulting in a solanesol enrichment yield of 87.33%. 1 g of the dried concentrate was added to 5 ml of ethyl acetate. The extraction temperature was 10°C for 3 hours. The extract was centrifuged at 6000 rpm for 6 minutes. The supernatant was removed and the ethyl acetate was spin-dried to obtain an ethyl acetate extract. 0.15 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract and saponified at 65°C for 2 hours. The saponified solution was neutralized with HCl and the pH was adjusted to approximately 7. The solution was centrifuged to obtain the supernatant, which was spin-dried to obtain a solanesol extract with a purity of 29.06%. 250 mg of the solanesol extract was dissolved in 1 ml of methanol and recrystallized at -20°C for 12 hours. The crystals were removed by centrifugation to obtain crude solanesol with a purity of 65.44%. 150 mg of crude solanesol was dissolved in 1 ml of ethanol solution, 50 ml of deionized water was added, and anti-solvent crystallization was performed. The solution was centrifuged to obtain pure solanesol. The purity of the pure solanesol was 91.21%.

[0060] Example 7:

[0061] Waste tobacco is crushed and sun-dried to obtain tobacco powder. One gram of tobacco powder is weighed and thoroughly mixed with 15 ml of 70 mg / ml sodium lauryl sulfate micelles to obtain a mixture. The mixture is then subjected to a microwave extraction device for extraction three times over a 5-minute period. After extraction, the mixture is centrifuged at 10,000 rpm for 7 minutes. The supernatant is collected to obtain a solanesol extract, which is then assayed for solanesol by high-performance liquid chromatography. The solanesol extraction yield is calculated to be 85.90%. A 2% by weight aqueous chitosan acetate solution is added to 100% of the solanesol extract. The chitosan acetate solution and the solanesol extract are stirred for 3 hours to obtain a mixed solution. The mixed solution is then centrifuged to obtain a solanesol concentrate, which is then dried. The solanesol enrichment yield is 100%. 1 g of the dried concentrate was added to 8 ml of ethyl acetate. The extraction temperature was 25°C for 2 hours. The extract was centrifuged at 7000 rpm for 10 minutes. The supernatant was removed and the ethyl acetate was spin-dried to obtain an ethyl acetate extract. 0.05 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract and saponified at 65°C for 3 hours. The saponified solution was neutralized with HCl and the pH was adjusted to approximately 7. The solution was centrifuged to obtain the supernatant, which was spin-dried to obtain a solanesol extract with a purity of 29.20%. 100 mg of the solanesol extract was dissolved in 1 ml of methanol and recrystallized at 10°C for 4 hours. The crystals were removed by centrifugation to obtain crude solanesol with a purity of 62.78%. 250 mg of crude solanesol was dissolved in 1 ml of ethanol solution, 30 ml of deionized water was added, and anti-solvent crystallization was performed. The solution was centrifuged to obtain pure solanesol. The purity of the pure solanesol was 88.05%.

[0062] Example 8:

[0063] Waste tobacco is crushed and sun-dried to obtain tobacco powder. One gram of tobacco powder is weighed and thoroughly mixed with 30 ml of 30 mg / ml sodium lauryl sulfate micelles to obtain a mixture. The mixture is subjected to ultrasonic extraction and extracted three times for 30 minutes. After extraction, the mixture is centrifuged at 9000 rpm for 6 minutes. The supernatant is collected to obtain a solanesol extract. The solanesol content in the extract is determined by high-performance liquid chromatography; the solanesol extraction yield is calculated to be 89.66%. A 3 wt% aqueous chitosan acetate solution is added to 50% of the volume of the solanesol extract. The chitosan acetate solution and the solanesol extract are stirred for 3 hours to obtain a mixed solution. The mixed solution is centrifuged to obtain a solanesol concentrate, which is then dried. The solanesol enrichment yield is 99.06%. 1 g of the dried concentrate was added to 9 ml of ethyl acetate. The extraction temperature was 10°C for 3 hours. The extract was centrifuged at 6000 rpm for 6 minutes. The supernatant was removed and the ethyl acetate was spin-dried to obtain an ethyl acetate extract. 0.2 mol / L sodium hydroxide ethanol solution was added to the ethyl acetate extract and saponified at 65°C for 4 hours. The saponified solution was neutralized with HCl and the pH was adjusted to approximately 7. The solution was centrifuged to obtain the supernatant, which was spin-dried to obtain a solanesol extract with a purity of 31.26%. 200 mg of the solanesol extract was dissolved in 1 ml of methanol and recrystallized at 0°C for 20 hours. The crystals were removed by centrifugation to obtain crude solanesol with a purity of 59.76%. 250 mg of crude solanesol was dissolved in 1 ml of ethanol solution, 30 ml of deionized water was added, and anti-solvent crystallization was performed. The solution was centrifuged to obtain pure solanesol. The purity of the pure solanesol was 90.66%.

[0064] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A process for extracting and purifying solanesol from waste and inferior tobacco leaves, specifically including a series of processes such as extraction with sodium dodecyl sulfate aqueous solution, flocculation separation with chitosan, extraction with ethyl acetate, saponification with sodium hydroxide, recrystallization, and purification by anti-solvent crystallization method, characterized in that, It includes the following steps: (1) Crush and dry the waste and inferior tobacco leaves to obtain tobacco leaf powder; (2) Mix the tobacco leaf powder with sodium dodecyl sulfate micelles at a certain solid-liquid ratio to obtain a mixture. Extract the mixture to obtain an extract. Centrifuge the extract and take the supernatant to obtain solanesol extract; (3) Add chitosan acetic acid aqueous solution to the solanesol extract, stir and mix to obtain a mixed solution. Centrifuge the mixed solution to obtain solanesol enrichment, and dry the enrichment; (4) Add ethyl acetate to the dried enrichment for extraction. Centrifuge the extraction solution and take the supernatant, and spin-dry the ethyl acetate to obtain ethyl acetate extract; (5) Add sodium hydroxide ethanol solution to the ethyl acetate extract for saponification. Neutralize the saponified solution with HCl, adjust the pH value to about 7. Centrifuge the solution to obtain the supernatant, and spin-dry to obtain solanesol extract; (6) Dissolve the solanesol extract in methanol solution, perform low-temperature recrystallization, centrifuge and take out the crystals to obtain crude solanesol; (7) Dissolve the crude solanesol in ethanol solution, add deionized water for anti-solvent crystallization. Centrifuge the solution to obtain pure solanesol.

2. The extraction and purification process of solanesol according to claim 1, characterized in that, In the step (2), the mass percentage content of sodium dodecyl sulfate micelles is 0.5-9 wt%; the mass ratio of the tobacco leaf powder to the volume of sodium dodecyl sulfate micelles is 1 g:10-35 ml; the extraction includes one or more of heating reflux extraction, microwave extraction, ultrasonic extraction or ultra-high pressure extraction; the number of heating reflux extractions is 1-3 times, and the extraction time is 30-60 min; the number of microwave extractions is 1-3 times, and the time is 5-30 min; the number of ultrasonic extractions is 1-3 times, and the time is 30-60 min; the number of ultra-high pressure extractions is 1-3 times, the extraction time is 0.5-3 min, and the extraction pressure is 50-300 MPa.

3. The extraction and purification process of solanesol according to claim 1, characterized in that, In the step (3), the mass percentage content of chitosan in the chitosan acetic acid aqueous solution is 0.5-3 wt%; the addition amount of the chitosan acetic acid aqueous solution is 50-100% of the volume of the solanesol extract, and the stirring and mixing time of the chitosan acetic acid aqueous solution and the solanesol extract is 1-3 h.

4. The extraction and purification process of solanesol according to claim 1, characterized in that, In the step (4), the mass ratio of the dried enrichment to the volume of the ethyl acetate extraction solution is 1 g:4-9 ml, the extraction temperature is 10-35 °C, and the extraction time is 1-3 h.

5. The extraction and purification process of solanesol according to claim 1, characterized in that, In the step (5), the concentration of the sodium hydroxide ethanol solution is 0.02-0.2 mol / L, the saponification time is 1-4 h, and the saponification temperature is 45-65 °C.

6. The extraction and purification process of solanesol according to claim 1, characterized in that, In the step (6), the concentration of the solanesol extract is 50-250 mg / ml, the recrystallization temperature is -20-20 °C, and the crystallization time is 4-20 h.

7. The extraction and purification process of solanesol according to claim 1, characterized in that, In the step (7), the concentration of the crude solanesol is 50-250 mg / ml, and the volume ratio of the solanesol solution for anti-solvent crystallization to deionized water is 1:10-1:

50.

8. The extraction and purification process of solanesol according to claim 1, characterized in that, In the steps (2)-(7), the centrifugation speed is 6000-10000 r / min, and the centrifugation time is 6-10 min.

Images