CN110590865A - Process method for extracting and purifying stevioside from stevia rebaudiana - Google Patents
Process method for extracting and purifying stevioside from stevia rebaudiana Download PDFInfo
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- CN110590865A CN110590865A CN201910947331.9A CN201910947331A CN110590865A CN 110590865 A CN110590865 A CN 110590865A CN 201910947331 A CN201910947331 A CN 201910947331A CN 110590865 A CN110590865 A CN 110590865A
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
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Abstract
The invention discloses a process method for extracting and purifying stevioside from stevia rebaudiana, which particularly relates to the technical field of plant extraction and comprises the following steps: step one, selecting a 800ml beaker, cleaning and sterilizing the beaker, pouring 200ml of deionized water into the sterilized beaker, heating and boiling the beaker, adding 20-30g of stevia rebaudiana dry leaves, and stirring the deionized water in the beaker by using a sterile glass rod. The invention carries out ultrahigh pressure treatment on the primarily cooked filtrate, extracts, can generate great osmotic pressure inside and outside raw material cells in the process of boosting pressure to promote a solvent to rapidly enter the cells through tiny channels of tissues such as cell walls and the like and fully contact with stevioside, and macromolecular substances such as proteins and the like can generate certain degree of denaturation and condensation by higher pressure, so that the structures of the cell walls and cell membranes become loose, the permeability of the cell walls is enhanced, the mass transfer resistance is greatly reduced, the leaching of the stevioside is facilitated, and the leaching rate of the stevioside is improved.
Description
Technical Field
The invention relates to the technical field of plant extraction, in particular to a process method for extracting and purifying stevioside from stevia rebaudiana.
Background
Stevia rebaudiana (Bertoni) Hemsl, a scientific name, is a perennial herb of the Stevia genus of the Compositae family. The plant height is 1-1.3 m. The root tips are large, 50-60 strips, and the length can reach 25 cm. Upright stem, lignified shoot at the base, tender upper part, dense short hairy hair, purple red or white corolla at the base and white upper part. The thin fruit is linear, slightly flat and brown and has crown hair. The flowering period is 7-9 months, and the fruit period is 9-11 months. Native to alpine grasslands bordering paraguay and brazil. The species prefers to grow in a warm and humid environment and is sensitive to light. Since 1977, China Beijing, Hebei, Shanxi, Jiangsu, Anhui, Fujian, Hunan, Yunnan and other places have been introduced for cultivation.
Stevia sugar is a mixture of diterpene glycosides containing 8 components extracted from leaves of stevia rebaudiana Bertoni, the leaves of stevia rebaudiana Bertoni contain 8% -16% of stevioside, and belongs to tetracyclic diterpene glycosides according to the division of natural phytochemistry, the sweetness of the stevioside is 200-300 times of that of cane sugar, the calorie of the stevioside is only 1/300 of cane sugar, the stevioside is gradually favored by people due to the characteristics of high sweetness, low calorie, safety, no toxicity, pure nature and the like, and the stevioside is widely applied to industries such as food, beverage, medicine, daily chemical industry and the like.
Among the known 9 glycosides of stevioside, the content, taste and sweetness of each ingredient are different, and among them, stevioside (stevioside), rebaudioside a (ra), and rebaudioside c (rc) are higher, accounting for more than 90%. RA has the highest sweetness, the sweetness characteristic of RA is similar to that of cane sugar, and the novel natural sweetener has the characteristics of high sweetness, low calorie, easy dissolution, heat resistance, stability and the like. It can be used for treating obesity, hypertension, and hyperglycemia. However, the existing extraction process of stevioside has the disadvantages of long extraction time, high water consumption, high organic wastewater generation amount and low product yield, and becomes an industrial pain point.
Disclosure of Invention
In order to overcome the above-mentioned defects of the prior art, the embodiment of the invention provides a process for extracting and purifying stevioside from stevia rebaudiana, which comprises the steps of performing ultrahigh pressure treatment on a primarily cooked filtrate, leaching, generating great osmotic pressure inside and outside raw material cells in the process of boosting pressure, promoting a solvent to rapidly enter the cells through tiny channels of tissues such as cell walls and the like, fully contacting with the stevioside, enabling macromolecular substances such as proteins and the like to generate certain degree of denaturation and condensation by higher pressure, loosening the structures of the cell walls and cell membranes, enhancing the permeability of the cell walls, greatly reducing mass transfer resistance, facilitating the leaching of the stevioside, improving the leaching rate of the stevioside, pressurizing and boiling the solution in a low-pressure environment, enabling the solution to complete evaporation at a lower temperature, completing the recovery of the solution, greatly reducing water consumption in the production process and reducing the generation of organic wastewater, and a purer solution is obtained, so that the purity and yield of the prepared product are higher, the final total yield of stevioside is more than 98%, and the method has obvious advantages compared with the conventional process.
In order to achieve the purpose, the invention provides the following technical scheme: a process for extracting purified stevioside from stevia rebaudiana Bertoni comprises the following steps:
step one, selecting an 800ml beaker, cleaning and disinfecting the beaker, pouring 200ml of deionized water into the disinfected beaker, heating and boiling, adding 20-30g of stevia rebaudiana dry leaves, stirring the deionized water in the beaker by using a sterile glass rod to enable the deionized water to completely submerge the stevia rebaudiana dry leaves, stewing for 30-50 minutes, cooling to room temperature, and filtering to remove scum and impurities in the solution to obtain filtrate;
step two, pouring the filtrate into a crucible, placing the crucible in an ultrahigh pressure environment, maintaining the pressure for 3-5 minutes, and then leaching the filtrate to obtain a leaching solution;
pouring the obtained leaching liquor into a beaker, adding deionized water with the mass ratio of 3:1 to the leaching liquor, heating to 80 ℃ in a low-pressure environment, preserving the heat for 40-60 minutes, leaching, and cooling the solution to room temperature in a normal-pressure environment to obtain a phase A;
pouring the obtained phase A into a beaker, stirring the solution at a constant speed by using a sterile glass rod, adding 3-5g of calcium oxide powder in the stirring process, sealing the beaker by using a preservative film after the calcium oxide powder is completely melted into the solution, standing for 1-3 hours at normal temperature, filtering the solution, and filtering scum and impurities in the solution to obtain a phase B;
step five, adding 2 mol. L into the B phase-1FeSO of (2)41-3ml of solution is uniformly stirred, the solution is sealed by a preservative film, placed in a normal temperature environment and kept stand for 1-3 hours, then the solution is placed in a low-pressure environment and heated to a boiling state and concentrated to 40% of the initial mass, then the solution is cooled to room temperature in a normal pressure environment, 50ml of ethanol solution is added, the solution is kept stand for 2-4 hours, filtered by an ultrafiltration membrane, adsorbed and separated by resin, and desalted and decolored to obtain a light yellow C phase;
sixthly, putting the phase C into a stainless steel tray, and drying in a freeze dryer for 6-8 hours to obtain light yellowDissolving stevioside in ethanol solution, and passing through anhydrous Na2SO4And (4) carrying out suction filtration, pouring the filtrate into a tray, and placing the tray in a ventilated place for natural cooling crystallization to obtain stevioside crystal powder.
In a preferred embodiment, in the second step, the ultrahigh pressure is specifically 100-.
In a preferred embodiment, in said third and fifth steps, the depression is in particular 4900 Pa.
In a preferred embodiment, in the fifth step, the volume fraction of the ethanol solution is 95%.
In a preferred embodiment, in the fourth step and the fifth step, the normal temperature is 15-25 ℃.
The invention has the technical effects and advantages that:
1. according to the invention, the filtrate subjected to preliminary cooking is subjected to ultrahigh pressure treatment and is extracted, so that during the pressure boosting process, great osmotic pressure can be generated inside and outside raw material cells to promote a solvent to rapidly enter the cells through micro channels of tissues such as cell walls and the like, the solvent is fully contacted with stevioside, and macromolecular substances such as proteins and the like can be denatured and condensed to a certain extent under higher pressure, so that the structures of the cell walls and cell membranes become loose, the permeability of the cell walls is enhanced, the mass transfer resistance is greatly reduced, the leaching of stevioside is facilitated, and the leaching rate of the stevioside is increased;
2. the solution is pressurized and boiled in a low-pressure environment, so that the solution can be evaporated at a lower temperature, the solution is recovered, the water consumption in the production process can be greatly reduced, the generation of organic wastewater is reduced, a purer solution is obtained, the purity and yield of the prepared product are higher, the final total yield of stevioside is more than 98%, and the stevioside preparation method has obvious advantages compared with the conventional process;
3. the method has the advantages of simple process, low equipment requirement, strong operability and good social popularization and application.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
a process for extracting purified stevioside from stevia rebaudiana Bertoni comprises the following steps:
step one, selecting an 800ml beaker, cleaning and disinfecting the beaker, pouring 200ml of deionized water into the disinfected beaker, heating and boiling, adding 20g of stevia rebaudiana dry leaves, stirring the deionized water in the beaker by using a sterile glass rod to enable the deionized water to completely submerge the stevia rebaudiana dry leaves, cooking for 30 minutes, cooling to room temperature, and filtering to remove scum and impurities in the solution to obtain filtrate;
step two, pouring the filtrate into a crucible, placing the crucible in an ultrahigh pressure environment of 100Mpa, maintaining the pressure for 3 minutes, and then leaching the filtrate to obtain a leaching solution;
pouring the obtained leaching liquor into a beaker, adding deionized water with the mass ratio of 3:1 to the leaching liquor, heating to 80 ℃ in a low-pressure environment of 4900Pa, preserving the temperature for 40 minutes, leaching, and then cooling the solution to room temperature in a normal-pressure environment to obtain a phase A;
pouring the obtained phase A into a beaker, stirring the solution at a constant speed by using a sterile glass rod, adding 3g of calcium oxide powder in the stirring process, sealing the beaker by using a preservative film after the calcium oxide powder is completely dissolved into the solution, standing for 1 hour at the temperature of 15 ℃, filtering the solution, and filtering scum and impurities in the solution to obtain a phase B;
step five, adding 2 mol. L into the B phase-1FeSO of (2)41ml of solution is evenly stirred, the solution is sealed by a preservative film, is placed at the temperature of 15 ℃ for standing for 1 hour, is placed in a low-pressure environment of 4900Pa to be heated to a boiling state and concentrated to 40 percent of the initial mass, and is circulated under normal pressureCooling to room temperature, adding 50ml of ethanol solution with volume fraction of 95%, standing for 2 hours, filtering with an ultrafiltration membrane, adsorbing and separating the solution with resin, and desalting and decolorizing the solution to obtain a light yellow C phase;
putting the phase C into a stainless steel tray, drying for 6 hours in a freeze dryer to obtain light yellow stevioside, dissolving the freeze-dried stevioside by using an ethanol solution, and passing anhydrous Na2SO4And (4) carrying out suction filtration, pouring the filtrate into a tray, and placing the tray in a ventilated place for natural cooling crystallization to obtain stevioside crystal powder.
Example 2:
a process for extracting purified stevioside from stevia rebaudiana Bertoni comprises the following steps:
step one, selecting an 800ml beaker, cleaning and disinfecting the beaker, pouring 200ml of deionized water into the disinfected beaker, heating and boiling, adding 25g of stevia rebaudiana dry leaves, stirring the deionized water in the beaker by using a sterile glass rod to enable the deionized water to completely submerge the stevia rebaudiana dry leaves, cooking for 40 minutes, cooling to room temperature, and filtering to remove scum and impurities in the solution to obtain filtrate;
step two, pouring the filtrate into a crucible, placing the crucible in an ultrahigh pressure environment of 110Mpa, maintaining the pressure for 4 minutes, and then leaching the filtrate to obtain a leaching solution;
pouring the obtained leaching liquor into a beaker, adding deionized water with the mass ratio of 3:1 to the leaching liquor, heating to 80 ℃ in a low-pressure environment of 4900Pa, preserving the temperature for 50 minutes, leaching, and then cooling the solution to room temperature in a normal-pressure environment to obtain a phase A;
pouring the obtained phase A into a beaker, stirring the solution at a constant speed by using a sterile glass rod, adding 4g of calcium oxide powder in the stirring process, sealing the beaker by using a preservative film after the calcium oxide powder is completely dissolved into the solution, standing for 2 hours at the temperature of 20 ℃, filtering the solution, and filtering scum and impurities in the solution to obtain a phase B;
step five, to phase B2 mol. L is added-1FeSO of (2)42ml of solution is uniformly stirred, the solution is sealed by a preservative film, the solution is placed at the temperature of 20 ℃ for standing for 2 hours, then the solution is placed at the low-pressure environment of 4900Pa for heating to a boiling state and concentrating to 40% of the initial mass, then the solution is cooled to room temperature under the normal-pressure environment, 50ml of ethanol solution with the volume fraction of 95% is added, the solution is placed for standing for 3 hours, after the solution is filtered by an ultrafiltration membrane, the solution is adsorbed and separated by resin, and the solution is desalted and decolored to obtain a light yellow C phase;
putting the phase C into a stainless steel tray, drying for 7 hours in a freeze dryer to obtain light yellow stevioside, dissolving the freeze-dried stevioside by using an ethanol solution, and passing anhydrous Na2SO4And (4) carrying out suction filtration, pouring the filtrate into a tray, and placing the tray in a ventilated place for natural cooling crystallization to obtain stevioside crystal powder.
Example 3:
a process for extracting purified stevioside from stevia rebaudiana Bertoni comprises the following steps:
step one, selecting an 800ml beaker, cleaning and disinfecting the beaker, pouring 200ml of deionized water into the disinfected beaker, heating and boiling the beaker, adding 30g of stevia rebaudiana dry leaves, stirring the deionized water in the beaker by using a sterile glass rod to enable the deionized water to completely submerge the stevia rebaudiana dry leaves, cooking the mixture for 50 minutes, cooling the mixture to room temperature, and filtering the mixture to remove scum and impurities in the solution to obtain filtrate;
step two, pouring the filtrate into a crucible, placing the crucible in an ultrahigh pressure environment of 120Mpa, maintaining the pressure for 5 minutes, and then leaching the filtrate to obtain a leaching solution;
pouring the obtained leaching liquor into a beaker, adding deionized water with the mass ratio of 3:1 to the leaching liquor, heating to 80 ℃ in a low-pressure environment of 4900Pa, preserving the temperature for 60 minutes, leaching, and then cooling the solution to room temperature in a normal-pressure environment to obtain a phase A;
pouring the obtained phase A into a beaker, stirring the solution at a constant speed by using a sterile glass rod, adding 5g of calcium oxide powder in the stirring process, sealing the beaker by using a preservative film after the calcium oxide powder is completely dissolved into the solution, standing for 3 hours at the temperature of 25 ℃, filtering the solution, and filtering scum and impurities in the solution to obtain a phase B;
step five, adding 2 mol. L into the B phase-1FeSO of (2)43ml of solution is uniformly stirred, the solution is sealed by using a preservative film, the solution is placed at the temperature of 25 ℃ and kept stand for 3 hours, then the solution is placed at the low-pressure environment of 4900Pa to be heated to a boiling state and concentrated to 40% of the initial mass, then the solution is cooled to room temperature under the normal-pressure environment, 50ml of ethanol solution with the volume fraction of 95% is added, the solution is kept stand for 4 hours, after being filtered by using an ultrafiltration membrane, the solution is adsorbed and separated by resin, and the solution is desalted and decolored to obtain a light yellow C phase;
putting the phase C into a stainless steel tray, drying for 8 hours in a freeze dryer to obtain light yellow stevioside, dissolving the freeze-dried stevioside by using an ethanol solution, and passing anhydrous Na2SO4And (4) carrying out suction filtration, pouring the filtrate into a tray, and placing the tray in a ventilated place for natural cooling crystallization to obtain stevioside crystal powder.
Three technological methods for extracting and purifying stevioside can be obtained through the three groups of embodiments, the three technological methods for extracting and purifying stevioside are respectively tested, and a common steaming method is used for testing, so that the leaching rates of the technological methods for extracting and purifying stevioside in the three groups of embodiments are improved differently, wherein the technological method for extracting and purifying stevioside in the embodiment 2 is the highest in technological method for extracting and purifying stevioside, the best in extraction effect, and various comparative data are as follows:
| leaching rate (%) | Water consumption (ml) | Consuming time (min) | |
| Example 1 | 98.42 | 282 | 34 |
| Example 2 | 98.96 | 269 | 27 |
| Example 3 | 98.63 | 291 | 39 |
| Comparative example 1 | 95.46 | 352 | 90 |
After the data are compared, the method is obviously superior to the existing cooking method, the filtrate subjected to preliminary cooking is subjected to ultrahigh pressure treatment and is extracted, in the boosting process, great osmotic pressure can be generated inside and outside raw material cells to promote a solvent to rapidly enter the cells through micro channels of tissues such as cell walls and the like, the solvent is fully contacted with stevioside, and high pressure can enable macromolecular substances such as proteins and the like to denature and agglomerate to a certain degree, so that the structures of the cell walls and the cell membranes become loose, the permeability of the cell walls is enhanced, the mass transfer resistance is greatly reduced, the method is favorable for leaching stevioside, and the leaching rate of the stevioside is improved;
through pressurizing and boiling the solution under the low-pressure environment, the solution can be evaporated at a lower temperature, the recovery of the solution is completed, the water consumption in the production process can be greatly reduced, the generation of organic wastewater is reduced, and purer solution is obtained, so that the purity and yield of the prepared product are higher, the final total yield of stevioside is more than 98%, and the method has obvious advantages compared with the conventional steaming and boiling method.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.
Claims (5)
1. A process method for extracting and purifying stevioside from stevia rebaudiana is characterized by comprising the following steps:
step one, selecting an 800ml beaker, cleaning and disinfecting the beaker, pouring 200ml of deionized water into the disinfected beaker, heating and boiling, adding 20-30g of stevia rebaudiana dry leaves, stirring the deionized water in the beaker by using a sterile glass rod to enable the deionized water to completely submerge the stevia rebaudiana dry leaves, stewing for 30-50 minutes, cooling to room temperature, and filtering to remove scum and impurities in the solution to obtain filtrate;
step two, pouring the filtrate into a crucible, placing the crucible in an ultrahigh pressure environment, maintaining the pressure for 3-5 minutes, and then leaching the filtrate to obtain a leaching solution;
pouring the obtained leaching liquor into a beaker, adding deionized water with the mass ratio of 3:1 to the leaching liquor, heating to 80 ℃ in a low-pressure environment, preserving the heat for 40-60 minutes, leaching, and cooling the solution to room temperature in a normal-pressure environment to obtain a phase A;
pouring the obtained phase A into a beaker, stirring the solution at a constant speed by using a sterile glass rod, adding 3-5g of calcium oxide powder in the stirring process, sealing the beaker by using a preservative film after the calcium oxide powder is completely melted into the solution, standing for 1-3 hours at normal temperature, filtering the solution, and filtering scum and impurities in the solution to obtain a phase B;
step five, adding 2 mol. L into the B phase-11-3ml of FeSO4 solution is uniformly stirred, the solution is sealed by a preservative film, placed in a normal temperature environment for standing for 1-3 hours, placed in a low-pressure environment for heating to a boiling state and concentrating to 40% of the initial mass, then cooled to room temperature in a normal pressure environment, 50ml of ethanol solution is added, then the solution is placed for standing for 2-4 hours, filtered by an ultrafiltration membrane, adsorbed and separated by resin, and desalted and decolored to obtain a light yellow C phase;
putting the phase C into a stainless steel tray, drying for 6-8 hours in a freeze dryer to obtain light yellow stevioside, dissolving the freeze-dried stevioside by using an ethanol solution, and passing anhydrous Na2SO4And (4) carrying out suction filtration, pouring the filtrate into a tray, and placing the tray in a ventilated place for natural cooling crystallization to obtain stevioside crystal powder.
2. The process of claim 1, wherein the extraction and purification of stevioside from stevia rebaudiana Bertoni comprises: in the second step, the ultrahigh pressure is specifically 100-120 Mpa.
3. The process of claim 1, wherein the extraction and purification of stevioside from stevia rebaudiana Bertoni comprises: in the third step and the fifth step, the low pressure is specifically 4900 Pa.
4. The process of claim 1, wherein the extraction and purification of stevioside from stevia rebaudiana Bertoni comprises: in the fifth step, the volume fraction of the ethanol solution is 95%.
5. The process of claim 1, wherein the extraction and purification of stevioside from stevia rebaudiana Bertoni comprises: in the fourth step and the fifth step, the normal temperature is 15-25 ℃.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111111256A (en) * | 2020-01-06 | 2020-05-08 | 江苏泽博环保科技有限公司 | Method and device for purifying plant water extracting solution |
| CN112167488A (en) * | 2020-06-29 | 2021-01-05 | 武汉林宝莱生物科技有限公司 | Cannabis plant beverage and preparation method thereof |
| CN113768015A (en) * | 2021-09-24 | 2021-12-10 | 尹帅 | Preparation method of keemun chrysanthemum water |
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2019
- 2019-10-08 CN CN201910947331.9A patent/CN110590865A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111111256A (en) * | 2020-01-06 | 2020-05-08 | 江苏泽博环保科技有限公司 | Method and device for purifying plant water extracting solution |
| CN112167488A (en) * | 2020-06-29 | 2021-01-05 | 武汉林宝莱生物科技有限公司 | Cannabis plant beverage and preparation method thereof |
| CN113768015A (en) * | 2021-09-24 | 2021-12-10 | 尹帅 | Preparation method of keemun chrysanthemum water |
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Application publication date: 20191220 |