CN106928023B - Metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco - Google Patents
Metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco Download PDFInfo
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- C07—ORGANIC CHEMISTRY
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Abstract
A metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco belongs to the technical field of solanesol extraction. According to the method, the tobacco solid waste is used as a raw material, the double synergistic catalysis effects of the compound enzyme for catalyzing and hydrolyzing the tobacco plant tissues and the manganese catalyst for improving the catalytic activity of the enzyme preparation are utilized, and the product is obtained through simple processes of raw material pretreatment, metal-enzyme preparation co-catalytic reaction, ultrasonic wave synergistic treatment and solanesol product preparation, so that the dissolution rate and the extraction rate of the product are improved. The method has the characteristics of wide raw material source, full waste utilization, low production cost, simple operation, environmental protection, high extraction rate and purity of the prepared solanesol product, complete biological activity and the like. The invention can be widely applied to the leaching and extraction of solanesol, nicotine, pectin, cellulose and other natural products extracted from tobacco solid waste, and effectively realizes the resource recycling of waste and inferior tobacco.
Description
Technical Field
The invention belongs to the technical field of solanesol extraction, and particularly relates to a metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco.
Background
Solanesol is triploid hemiterpene alcohol, naturally exists in tobacco leaves (about 0.3-3% of the total weight), potato leaves (about 1.5%), mulberry leaves (about 0.1%) and the like, has strong anticancer biological activity, is often used as an important raw material for drug synthesis, and is applied to: synthesizing ubiquinone compound such as coenzyme Q10; ② synthesizing a medicament for treating tumor; synthesizing antiulcer medicine; synthesizing the medicine for treating cardiovascular and cerebrovascular diseases; fifthly, synthesizing the anti-HIV drug, and having wide application prospect. The existing solanesol extraction technology mainly comprises a solvent extraction method, a supercritical extraction method, a polymerization state coprecipitation method, a fresh method homogenate extraction method, a molecular distillation technology and the like, can more or less effectively degrade lignocellulose to promote the dissolution of natural solanesol, but the process conditions of strong acid (alkali), high temperature (pressure) and the like not only cause high energy consumption and high pollution, but also easily cause the addition, oxidation, dehydrogenation, molecular rearrangement reaction and the like of the solanesol, and reduce the resource utilization rate. Therefore, the research on improving the extraction method of the solanesol has very important significance.
The existing method for extracting solanesol, for example, "a process for extracting solanesol" with publication number CN104072336A, disclosed on 10.1.2014, uses waste and inferior tobacco leaves as raw materials, adopts a supercritical chromatography, utilizes good selectivity of supercritical carbon dioxide and selectivity of resin, and obtains the solanesol by raw material pretreatment, primary extraction, secondary extraction, tertiary extraction, recrystallization after elution, filtration and drying. Wherein, the first-stage extraction is crude extraction, all components in the waste and inferior tobacco leaves are leached by supercritical carbon dioxide, the filling rate of the raw materials is 80 percent, and the extraction temperature is 65-75 ℃; performing secondary extraction by using adsorbent (molecular sieve, aluminum oxide or active carbon) in an extraction kettle to complete dewaxing, degreasing and impurity removal at 60-70 ℃; the third-stage extraction uses selective chromatographic silica gel or resin to complete the adsorption of solanesol, and adopts ethyl acetate or petroleum ether to elute so as to obtain the solanesol with the purity of 95%. The main disadvantages of this method are: (1) the method utilizes supercritical carbon dioxide to leach solanesol in the tobacco leaves, but simultaneously enhances the leaching of other components in the tobacco leaves, and adopts three-stage extraction procedures because the impurity removal difficulty is increased due to the diversity of the leached components, thereby increasing production equipment and increasing production cost; (2) the method adopts adsorbents with different functions at each stage of three-stage extraction, increases production raw materials and increases production cost; (3) in the secondary extraction, a molecular sieve with adsorption function, aluminum oxide or active carbon is used as an adsorbent, and although dewaxing, degreasing and impurity removal are completed, solanesol can be adsorbed, so that the extraction rate of the solanesol in the tertiary extraction is reduced, and the production yield is reduced; (4) the method needs supercritical technology, and the extraction temperature is higher than 60 ℃ in the process, so that special production equipment is needed, and additional heat source equipment is needed, and the production cost is further increased.
Disclosure of Invention
The invention aims to provide a metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco aiming at the defects of the existing solanesol extraction method, and has the characteristics of mild reaction conditions, few and simple operation steps, sufficient solanesol leaching, few impurities, low production energy consumption, low cost and the like.
The mechanism of the invention is as follows: the method takes waste and inferior tobacco as a raw material, selects engineering enzyme (xylanase: cellulase: laccase) as an enzyme preparation for catalyzing and activating lignocellulose and manganese catalyst as an activator of the enzyme preparation, degrades lignin of the powdered cellulose in the tobacco through the xylanase and the laccase, increases the hydrolysis effect of the cellulase on the cellulose, improves the catalytic activity of the enzyme preparation through a metal catalyst, enhances the hydrolysis degree of tobacco plant fiber tissue which hinders the dissolution of the solanesol, improves the permeability of the plant tissue, reduces the mass transfer resistance of the solanesol, and increases the extraction rate of the solanesol. The two are used for catalyzing and hydrolyzing the plant fibrous tissue of the tobacco in a synergistic manner, so that the dissolution of other active ingredients in the tobacco is reduced due to the specificity of enzyme, the use amount of acid and alkali of chemical activators is avoided, the potential safety hazard of solanesol production is reduced, the secondary pollution to the environment caused by the increase of the use amount of acid and alkali is avoided, the environmental protection is facilitated, and the resource recycling of waste tobacco is realized.
The technical scheme for realizing the purpose of the invention is as follows: a metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco is characterized in that the waste and inferior tobacco is used as a raw material, and a product is prepared by raw material pretreatment, metal-enzyme preparation co-catalysis reaction, ultrasonic wave co-treatment and solanesol product preparation. The method comprises the following specific steps:
(1) pretreatment of raw materials
Taking tobacco solid waste (including waste and inferior tobacco leaves, tobacco stems, leftovers in the tobacco processing process, tobacco dregs for extracting total natural products of tobacco and the like) as raw materials. Cleaning the raw materials with water, placing the cleaned raw materials in an oven, drying the cleaned raw materials in the oven at 85-110 ℃ for 30-90 min, crushing the dried raw materials by a crusher, sieving the crushed raw materials by a 20-120-mesh sieve, returning the un-sieved raw materials to the crusher, crushing the crushed raw materials, and collecting the sieved raw materials (namely the pretreated raw materials).
(2) Metal-enzyme preparation co-catalyzed reaction
After the step (1) is finished, the mass (g) of the raw materials treated in the step (1) to the mass (g) of the catalyst are firstly) Adding a catalyst and water into the raw material pretreated in the step (1) according to the volume (ml) ratio of 1: 0.0125-0.075: 5-15, uniformly stirring, adjusting the pH value of the system to 4-8 by using sulfuric acid or a dilute sodium hydroxide solution, placing the mixed solution after adjusting the pH value in a shaking table, carrying out enzymolysis and activation for 4-8 h at the water bath temperature of 20-45 ℃, carrying out suction filtration by using a suction filtration pump, and respectively collecting filtrate and filter residue. Collecting the filtrate as water phase extractive solution containing solanesol; the collected filter residue is the tobacco base material after catalytic activation by the catalyst. The catalyst is a metal-enzyme preparation of metal and compound enzyme, and the adding amount ratio of the metal catalyst to the enzyme preparation is (g) =1: 1.6-10; the metal is manganese zinc ferrite (Mn)0.3Zn0.2Fe0.5Ox) Or Mn2+Or Fe3+(ii) a The enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase = 2-14: 23-47: 51-64.
(3) Ultrasonic wave co-processing
After the step (2) is completed, transferring the filter residue collected in the step (2), namely the tobacco base material, into an extraction container, adding the composite extracting agent solution into the extraction container according to the ratio of the mass (g) of the raw material pretreated in the step (1) to the volume (ml) of the composite extracting agent solution of 1: 8-12, stirring and mixing uniformly, carrying out ultrasonic treatment for 5-40 min under the conditions of the ultrasonic power of 100-200W and the temperature of 30-55 ℃, then carrying out suction filtration by using a suction filtration machine, and respectively collecting filtrate and filter residue. The collected filter residue is used as a raw material for preparing an adsorption material; transferring the collected filtrate into a centrifuge, performing centrifugal separation for 10-40 min under the condition that the centrifugal rotating speed is 3000-4000 r/min, and respectively collecting centrifugal clear liquid (namely the solanesol-containing extracting solution) and centrifugal slag. Transferring the collected centrifugal slag into the filter residue obtained in the step (2) to be used as a raw material for preparing the adsorbing material; and (3) determining the concentration of the solanesol in the collected centrifugal clear liquid by using a high performance liquid chromatography, namely determining the mass concentration and the dissolution rate of the solanesol in the collected centrifugal clear liquid under the conditions that the flow rate is 0.6-1.2 ml/min and the column temperature is 20-30 ℃ according to the volume ratio of mobile phase ethanol to methanol of the high performance liquid chromatography being 1: 4-9. The dissolution rate of the solanesol is 92.00-98.70%, and the dissolution rate is improved by 15.00-24.90%. The composite extractant solution is a mixed solution of ethanol and methanol with the mass concentration ratio of 1: 1-9.
(4) Preparation of solanesol product
And (3) after the step (3) is finished, placing the centrifugal clear liquid collected in the step (3) in a rotary evaporator, and carrying out reduced pressure concentration to obtain a viscous crude solanesol product under the vacuum pressure of 0.1-0.6 Mpa and the temperature of 30-60 ℃. Then, according to the ratio of the mass (g) of the crude solanesol product to the volume (ml) of the compound extractant solution being 1: 3-6, adding the crude solanesol product and the compound extractant solution into an extraction container, and mixing and dissolving uniformly. And finally, performing reduced pressure concentration for 2-4 times through a rotary evaporator under the vacuum pressure of 0.1-0.6 Mpa and the temperature of 30-60 ℃ to obtain the solanesol product. The extraction rate of the solanesol reaches 90.0-97.5%, and the purity reaches 91.0-94.0%.
After the technical scheme is adopted, the invention mainly has the following effects:
(1) the method selects the metal catalyst and the compound enzyme as the synergistic activator for catalyzing and activating the lignocellulose, and improves the extraction rate of the solanesol. And the tobacco lignocellulose is degraded by the metal-enzyme preparation under the synergetic catalysis, so that the biological activity of the enzyme preparation can be improved, the internal permeability of the tobacco is further improved, the dissolution mass transfer resistance of the solanesol is reduced, and the solanesol is fully dissolved and leached, therefore, the dissolution rate of the solanesol obtained by the method is 98.64 percent, which is improved by 24.90 percent compared with the ultrasonic alcohol extraction method, and the extraction rate can reach 97.3 percent.
(2) The invention uses metal-enzyme preparation to replace the traditional chemical activator acid-base to leach the solanesol and the nicotine, reduces the damage of chemical reagents such as acid-base and the like to the biological activity of the solanesol, has less interference components in the leaching liquor, completely retains the biological activity and can ensure the product quality. The enzyme is used for replacing acid and alkali of the activating agent, and meanwhile, the corrosion of the acid and alkali to production equipment in the production process is avoided, so that the service life of the production equipment is shortened, the environmental protection is facilitated, and the production cost is reduced.
(3) The method adopts the tobacco solid waste, the catalytic activation condition of enzyme (cellulase or lignin or a compound enzyme of the cellulase and the lignin enzyme) is optimal at 20-40 ℃, the enzymolysis time is only 7.5 hours, the reaction condition is mild, the operation is simple, and the organic solvent used for extracting the solanesol is methanol and ethanol, so the production potential safety hazard of extracting the solanesol by high temperature or a large amount of toxic organic solvents is reduced, the environmental management cost in the production process is reduced, the production safety performance is improved, and the energy consumption is reduced.
(4) The method has the advantages of wide raw material source, full utilization of waste, low production cost, simple operation and environmental protection. The method can be widely applied to extracting the solanesol by taking the solid tobacco waste as the raw material, and can also be widely applied to extracting nicotine, pectin and cellulose from the solid tobacco waste.
Detailed Description
The present invention will be further described with reference to the following embodiments.
Example 1
A metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco comprises the following specific steps:
(1) pretreatment of raw materials
Cleaning 40g of waste tobacco leaves, airing, placing in an oven, drying in the oven at 100 ℃ for 30min, taking out, crushing by a crusher, sieving by a 60-mesh molecular sieve to obtain the tobacco leaves with the particle size of less than 60 meshes, and subpackaging by plastic bags for later use.
(2) Metal-enzyme preparation co-catalyzed reaction
After the step (1) is finished, firstly, according to the ratio of the mass (g) of the raw materials treated in the step (1) to the mass (g) of the catalyst to the volume (ml) of water being 1: 0.035: 10, adding the catalyst and the water into the raw materials pretreated in the step (1), uniformly stirring, then adjusting the pH value of the system to 7 by using sulfuric acid or a dilute sodium hydroxide solution, then placing the mixed solution after the pH value is adjusted in a shaking table, carrying out enzymolysis and activation for 7.5 hours at the water bath temperature of 45 ℃, carrying out suction filtration by using a suction filtration pump, and respectively collecting filtrate and filter residues. Collecting the filtrate as water phase extractive solution containing solanesol; the collected filter residue is a catalystCatalytically activated tobacco base material. The catalyst is a metal-enzyme preparation of metal and compound enzyme, and the adding amount ratio of the metal catalyst to the enzyme preparation is (g) =1: 5; the metal is manganese zinc ferrite Mn0.3Zn0.2Fe0.5Ox(ii) a The enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase = 14: 23: 63.
(3) Ultrasonic wave co-processing
After the step (2) is finished, firstly transferring the filter residue collected in the step (2), namely the tobacco base material, into a leaching container, then adding the composite extracting agent solution into the leaching container according to the ratio of the mass (g) of the raw material pretreated in the step (1) to the volume (ml) of the composite extracting agent solution of 1:10, stirring and mixing uniformly, carrying out ultrasonic treatment for 25min under the conditions of ultrasonic power of 200W and temperature of 50 ℃, carrying out suction filtration by using a suction filtration machine, and respectively collecting filtrate and filter residue. The collected filter residue is used as a raw material for preparing an adsorption material; transferring the collected filtrate into a centrifuge, performing centrifugal separation for 15min under the condition that the centrifugal rotation speed is 4000r/min, and respectively collecting centrifugal clear liquid (namely the extracting solution containing the solanesol) and centrifugal slag. Transferring the collected centrifugal slag into the filter residue obtained in the step (2) to be used as a raw material for preparing the adsorbing material; and (3) determining the concentration of the solanesol in the collected centrifugal clear liquid by high performance liquid chromatography, namely determining the mass concentration and the dissolution rate of the solanesol in the collected centrifugal clear liquid according to the volume ratio of mobile phase ethanol to methanol of the high performance liquid chromatography of 1: 6 under the conditions that the flow rate is 0.8ml/min and the column temperature is 25 ℃. The solanesol leaching rate is 98.64 percent, and the leaching rate is improved by 24.9 percent. The composite extractant solution is a mixed solution of ethanol and methanol with the mass concentration ratio of 1: 5.5.
(4) Preparation of solanesol product
And (3) after the step (3) is finished, putting the centrifugal clear liquid collected in the step (3) into a rotary evaporator, and carrying out reduced pressure concentration to obtain a viscous crude solanesol product under the vacuum pressure of 0.2Mpa and the temperature of 45 ℃. Then, according to the ratio of the mass (g) of the crude product of the solanesol to the volume (ml) of the compound extractant solution being 1:5, the crude product of the solanesol and the compound extractant solution are added into an extraction container and mixed and dissolved evenly. And finally, performing reduced pressure concentration for 3 times through a rotary evaporator under the vacuum pressure of 0.2Mpa and the temperature of 45 ℃ to obtain the solanesol product. The extraction rate of solanesol is up to 97.3%, and the purity is up to 93.5%.
Example 2
A method for improving the co-catalysis of a metal-enzyme preparation for solanesol leaching in waste and inferior tobacco, which is the same as in example 1, wherein:
in the step (1), the tobacco mesh number is 20 meshes, the oven temperature is 85 ℃, and the drying time is 90 min.
In the step (2), the ratio of the mass (g) of the tobacco solid waste to the mass (g) of the catalyst to the volume (ml) of water is 1: 0.0125: 5. The pH value of the system is adjusted to be 4 by adopting a hydrochloric acid solution, and the temperature of the enzymolysis water bath is 25 ℃. And (4) carrying out enzymolysis activation on the tobacco base material for 8 hours. The adding amount ratio of the metal-enzyme preparation is metal catalyst (g) to enzyme preparation (g) =1: 1.6; the enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase = 2: 47: 51 built enzyme.
In the step (3), the mass ratio of the pretreated tobacco solid waste to the volume ratio of the composite extracting agent solution is 1:8, and the volume ratio of the composite extracting agent ethanol to the methanol is 1:1. The ultrasonic power is 200W, the ultrasonic temperature is 30 ℃, and the ultrasonic time is 5 min; the centrifugation revolution of the filtrate is 4000r/min, and the centrifugation time is 25 min. The volume ratio of mobile phase ethanol and methanol of high performance liquid chromatography is 1: 9, the flow rate is 1.2ml/min, and the column temperature is 30 ℃. The solanesol dissolution rate is 92.03 percent, and the dissolution rate is improved by 16.56 percent.
In the step (4), the vacuum pressure of the vacuum filtration is 0.6Mpa, and the concentration temperature is 60 ℃. The ratio of the mass (g) of the crude product of solanesol to the volume (ml) of the compound extractant solution is 1: 3. The extraction rate of solanesol is up to 95.9%, and the purity is up to 91.5%.
Example 3
A method for improving the co-catalysis of a metal-enzyme preparation for solanesol leaching in waste and inferior tobacco, which is the same as in example 1, wherein:
in the step (1), the tobacco with the mesh number of 120 meshes is selected, the temperature of an oven is 110 ℃, and the drying time is 30 min.
In the step (2), the ratio of the mass (g) of the tobacco solid waste to the mass (g) of the catalyst to the volume (ml) of water is 1: 0.075: 15. The pH value of the system is adjusted to be 6 by adopting a hydrochloric acid solution, and the water bath temperature is 45 ℃. And (4) activating the tobacco base material by enzymolysis for 4 hours. The adding amount ratio of the metal-enzyme preparation is metal catalyst (g) to enzyme preparation (g) =1: 10; the enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase = 2: 34: 64 formulated enzyme.
In the step (3), the mass ratio of the pretreated tobacco solid waste to the volume ratio of the composite extracting agent solution is 1:12, and the volume ratio of the composite extracting agent ethanol to the methanol is 1: 9. The ultrasonic power is 100W, the ultrasonic temperature is 55 ℃, and the ultrasonic time is 40 min; the centrifugation revolution of the filtrate is 3000r/min, and the centrifugation time is 10 min. The volume ratio of mobile phase ethanol and methanol of high performance liquid chromatography is 1: 4, the flow rate is 0.6ml/min, and the column temperature is 20 ℃. The solanesol leaching rate is 95.30 percent, and the leaching rate is improved by 20.7 percent.
In the step (4), the vacuum pressure of the vacuum filtration is 0.1Mpa, and the concentration temperature is 30 ℃. The ratio of the mass (g) of the crude product of solanesol to the volume (ml) of the compound extractant solution is 1: 6. The solanesol extraction rate reaches 93.2 percent, and the purity reaches 92.0 percent.
Results of the experiment
1. Comparison of the Process of example 1 with other Processes for the Leaching of Solanesol
TABLE 1 comparison of solanesol extraction product with the complex enzymatic hydrolysis and sonoethanol extraction process
| Method of producing a composite material | Concentration (mg/ml) | Leaching rate (%) | Leaching improvement (%) | Extraction ratio (%) | Purity (%) | Amount of organic solvent used (ml) | Reaction temperature (. degree.C.) | Extraction time (min) |
| Metal-enzyme preparation decomposing process | 0.1794 | 98.64 | 24.90 | 97.30 | 93.50 | 20 | 45 | 30 |
| Complex enzyme catalytic hydrolysis | 0.1687 | 92.76 | 17.49 | 92.80 | 91.0 | 20 | 45 | 30 |
| Ultrasonic alcohol extraction | 0.1435 | 78.95 | __ | 78.95 | 75.60 | 50 | 50. Heating of | 50 |
From the above experiments it is known that: the method adopts metal-enzyme co-catalysis activation to treat the tobacco solid waste to prepare the activated carbon, and when the mass ratio of the added tobacco waste to the solid tobacco waste is 1: 0.035 (xylanase: cellulase: laccase enzyme activity ratio 14: 23: 63), adding water with volume ratio of 1:10, at 45 ℃ in a water bath at the temperature close to room temperature, the enzymolysis time is 7.5h, the solanesol product can be prepared by the synergistic ultrasonic treatment of the compound extractant, and the by-product nicotine can be obtained at the same time. Wherein the ratio of the mass (g) of the tobacco enzymolysis activation base material to the volume (ml) of the compound extractant solution is 1:10, and the weight ratio of the compound extractant ethanol: the volume ratio of methanol is 1: 5.5, the ultrasonic power is 200W, the ultrasonic temperature is 50 ℃, and the ultrasonic extraction time is 25 min. The method can fully dissolve out solanesol, and the leaching rate is 98.64 percent, which is 24.90 percent higher than that of the ultrasonic alcohol extraction method. The method has the advantages that the enzyme replaces an acid-base activating agent, the preparation conditions are mild, the corrosion of acid and base to production equipment is avoided, the potential safety hazard of high-temperature prepared products is reduced, the operation is simple, the extraction rate and purity of solanesol products are high, the biological activity is complete, the secondary pollution of acid, base and harmful organic matters to the environment in the production process is reduced, the environmental protection is facilitated, and the production cost is reduced.
Claims (4)
1. A metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco is characterized by comprising the following specific steps:
(1) pretreatment of raw materials
The method comprises the following steps of taking solid tobacco waste as raw materials, wherein the solid tobacco waste comprises waste inferior tobacco leaves, tobacco stems, leftovers in the tobacco processing process and tobacco residues for extracting total natural products of tobacco, cleaning the raw materials with water, placing the cleaned raw materials into an oven, drying the cleaned raw materials in the oven at 85-110 ℃ for 30-90 min, crushing the dried raw materials by a crusher, sieving the crushed raw materials by a sieve with 20-120 meshes, returning the unsieved raw materials to the crusher for crushing, and collecting the sieved raw materials, namely the pretreated raw materials;
(2) metal-enzyme preparation co-catalyzed reaction
After the step (1) is finished, according to the volume ratio of the mass of the raw material to the mass of the catalyst to the water after the step (1) is 1g to 0.0125-0.075 g to 5-15 ml, adding a catalyst and water into the raw material pretreated in the step (1), uniformly stirring, adjusting the pH value of the system to 4-8 by using sulfuric acid or a dilute sodium hydroxide solution, then placing the mixed solution after the pH value is adjusted in a shaking table, carrying out enzymolysis activation for 4-8 h at the water bath temperature of 20-45 ℃, carrying out suction filtration by using a suction filtration pump, respectively collecting filtrate and filter residue, wherein the collected filtrate is a water phase extracting solution containing solanesol, the collected filter residue is a tobacco base material which is catalyzed and activated by a catalyst, the catalyst is a metal-enzyme preparation of metal and compound enzyme, and the adding amount ratio of the metal catalyst to the enzyme preparation is 1g: 1.6-10 g; the metal is manganese zinc ferrite Mn0.3Zn0.2Fe0.5Ox(ii) a The enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase is 2-14: 23-47: 51-64 of compound enzyme;
(3) ultrasonic wave co-processing
After the step (2) is completed, firstly transferring the filter residue collected in the step (2), namely the tobacco base material, into an extraction container, then adding the composite extracting agent solution into the extraction container according to the ratio of the mass of the raw material pretreated in the step (1) to the volume of the composite extracting agent solution of 1g: 8-12 ml, stirring and mixing uniformly, carrying out ultrasonic treatment for 5-40 min under the conditions of the ultrasonic power of 100-200W and the temperature of 30-55 ℃, then carrying out suction filtration by using a suction filtration machine, respectively collecting filtrate and filter residue, and taking the collected filter residue as the raw material for preparing the adsorbing material; transferring the collected filtrate into a centrifugal machine, performing centrifugal separation for 10-40 min under the condition that the centrifugal rotating speed is 3000-4000 r/min, respectively collecting centrifugal clear liquids, namely the extracting solution containing the solanesol and the centrifugal slag, transferring the collected centrifugal slag into the filter residue obtained in the step (2), and taking the filter residue as a raw material for preparing the adsorbing material; determining the concentration of solanesol in the collected centrifugal clear liquid by a high performance liquid chromatography, namely determining the mass concentration and the dissolution rate of the solanesol in the collected centrifugal clear liquid according to the volume ratio of mobile phase ethanol to methanol of the high performance liquid chromatography of 1: 4-9 under the conditions that the flow rate is 0.6-1.2 ml/min and the column temperature is 20-30 ℃, wherein the dissolution rate of the solanesol is 92.00-98.70%, and the composite extractant solution is a mixed solution of ethanol and methanol with the mass concentration ratio of ethanol to methanol of 1: 1-9;
(4) preparation of solanesol product
After the step (3) is completed, the centrifugal clear liquid collected in the step (3) is placed in a rotary evaporator, vacuum concentration is carried out at the vacuum pressure of 0.1-0.6 Mpa and the temperature of 30-60 ℃ until the centrifugal clear liquid is viscous, a crude solanesol product is obtained, then the crude solanesol product and the composite extractant solution are added into an extraction container according to the ratio of the mass of the crude solanesol product to the volume of the composite extractant solution of 1g: 3-6 ml, the mixture is uniformly dissolved, finally, the rotary evaporator is used for carrying out vacuum concentration for 2-4 times at the vacuum pressure of 0.1-0.6 Mpa and the temperature of 30-60 ℃, and the solanesol product is obtained, wherein the solanesol extraction rate is 90.0-97.5%, and the purity is 91.0-94.0%.
2. The metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco as claimed in claim 1, wherein:
in the step (1), the waste tobacco with the mesh number of 60 meshes is selected, the temperature of an oven is 100 ℃, and the drying time is 30 min;
in the step (2), the ratio of the mass of the tobacco solid waste to the mass of the catalyst to the volume of water is 1g: 0.035 g:10 ml, the pH value of the solution is 7, the temperature of the enzymolysis water bath is 45 ℃, the enzymolysis time is 7.5h, the ratio of manganese, zinc and ferrite in the catalyst to the enzyme preparation is 1g: 5g, and the enzyme activity ratio of the enzyme preparation is xylanase: cellulase: laccase 14: 23: 63;
in the step (3), the mass ratio of the pretreated tobacco solid waste to the volume of the composite extractant solution is 1:10, the volume ratio of the composite extractant ethanol to the methanol is 1: 5.5, the ultrasonic power is 200W, the ultrasonic temperature is 50 ℃, and the ultrasonic time is 25 min; centrifuging the filtrate at 4000r/min for 15min, wherein the volume ratio of ethanol to methanol in mobile phase of high performance liquid chromatography is 1: 6, the flow rate is 0.8ml/min, the column temperature is 25 deg.C, and the solanesol dissolution rate is 98.64%;
in the step (4), the vacuum pressure of the vacuum pump filtration is 0.2Mpa, the concentration temperature is 45 ℃, the ratio of the mass of the crude product of the solanesol to the volume of the compound extractant solution is 1g to 5ml, the extraction rate of the solanesol reaches 97.3 percent, and the purity reaches 93.5 percent.
3. The metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco as claimed in claim 1, wherein:
in the step (1), the tobacco mesh number is 20 meshes, the temperature of an oven is 85 ℃, and the drying time is 90 min;
in the step (2), the ratio of the mass of the tobacco solid waste to the mass of the catalyst to the volume of water is 1: 0.0125 g: 5ml, the pH value of a system is adjusted to be 4 by adopting a hydrochloric acid solution, the water bath temperature is 25 ℃, the tobacco base material is activated by enzymolysis for 8 hours, and the adding amount ratio of the metal-enzyme preparation is that the metal catalyst to the enzyme preparation is 1g:1.6 g; the enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase is 2: 47: 51 complex enzyme;
in the step (3), the mass ratio of the pretreated tobacco solid waste to the volume of the composite extractant solution is 1:8, the volume ratio of the composite extractant ethanol to the methanol is 1:1, the ultrasonic power is 200W, the ultrasonic temperature is 30 ℃, and the ultrasonic time is 5 min; centrifuging the filtrate at 4000r/min for 25min, wherein the volume ratio of ethanol to methanol in mobile phase of high performance liquid chromatography is 1: 9, the flow rate is 1.2ml/min, the column temperature is 30 deg.C, and the solanesol dissolution rate is 92.03%;
4. the metal-enzyme preparation co-catalysis method for improving solanesol leaching in waste and inferior tobacco as claimed in claim 1, wherein:
in the step (1), the tobacco with the mesh number of 120 meshes is selected, the temperature of an oven is 110 ℃, and the drying time is 30 min;
in the step (2), the ratio of the mass of the tobacco solid waste to the mass of the catalyst to the volume of water is 1g to 0.075g to 15ml, the pH value of a system is adjusted to 6 by adopting a hydrochloric acid solution, the water bath temperature is 45 ℃, the tobacco base material is activated by enzymolysis for 4 hours, and the ratio of the metal catalyst to the enzyme preparation is 1g to 10 g; the enzyme preparation has the enzyme activity ratio of xylanase: cellulase: laccase is 2: 34: 64;
in the step (3), the mass ratio of the pretreated tobacco solid waste to the volume of the composite extractant solution is 1:12, the volume ratio of the composite extractant ethanol to the methanol is 1: 9, the ultrasonic power is 100W, the ultrasonic temperature is 55 ℃, and the ultrasonic time is 40 min; centrifuging the filtrate at 3000r/min for 10min at a flow rate of 0.6ml/min and a column temperature of 20 deg.C, wherein the volume ratio of ethanol to methanol in mobile phase of high performance liquid chromatography is 1: 4, the dissolution rate of solanesol is 95.30%;
in the step (4), the vacuum pressure of the vacuum pump filtration is 0.1Mpa, the concentration temperature is 30 ℃, the ratio of the mass of the crude product of the solanesol to the volume of the compound extractant solution is 1g to 6ml, the extraction rate of the solanesol reaches 93.2 percent, and the purity reaches 92.0 percent.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102816050A (en) * | 2012-09-13 | 2012-12-12 | 王星敏 | Method for extracting solanesol by activating waste and defective tobacco by enzyme catalysis |
| CN103014099A (en) * | 2012-12-27 | 2013-04-03 | 山东大学 | Method for promoting hydrolysis of lignocellulose |
| WO2014009749A1 (en) * | 2012-07-13 | 2014-01-16 | Kind Consumer Limited | Process for using tobacco |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014009749A1 (en) * | 2012-07-13 | 2014-01-16 | Kind Consumer Limited | Process for using tobacco |
| CN102816050A (en) * | 2012-09-13 | 2012-12-12 | 王星敏 | Method for extracting solanesol by activating waste and defective tobacco by enzyme catalysis |
| CN103014099A (en) * | 2012-12-27 | 2013-04-03 | 山东大学 | Method for promoting hydrolysis of lignocellulose |
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| 纤维素酶、酯酶和木聚糖酶对玉米秸秆协同作用的研究;宋金利;《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》;20120115(第01期);第1.2.7节，正文第48页第1段 * |
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