CN114805456A - Method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis - Google Patents
Method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis Download PDFInfo
- Publication number
- CN114805456A CN114805456A CN202210275100.XA CN202210275100A CN114805456A CN 114805456 A CN114805456 A CN 114805456A CN 202210275100 A CN202210275100 A CN 202210275100A CN 114805456 A CN114805456 A CN 114805456A
- Authority
- CN
- China
- Prior art keywords
- rebaudioside
- alkaline hydrolysis
- purity
- preparing
- acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/256—Polyterpene radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention discloses a method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis, which comprises the following steps: selecting rebaudioside a, adding water to obtain a rebaudioside a aqueous solution, adding a base into the rebaudioside a aqueous solution to adjust the pH value to 9-12, reacting at a constant temperature of 60-100 ℃ for 2-24 hours, adding an acid to neutralize after the completion of the alkaline hydrolysis reaction, adjusting the pH value to 6-8, standing the neutralized reaction liquid, separating out a rebaudioside b crude product, and performing solid-liquid separation and drying on the rebaudioside b crude product to prepare rebaudioside b. The rebaudioside b is prepared by selectively hydrolyzing the glycosidic bond of C19 in rebaudioside a without destroying the glycosidic bond of C13, the rebaudioside a is prepared by a basic hydrolysis method, the converted rebaudioside b is easy to separate out, and the subsequent separation and purification are simple and efficient.
Description
Technical Field
The invention belongs to the technical field of stevioside, and particularly relates to a method for preparing high-purity rebaudioside b by alkaline hydrolysis of rebaudioside a.
Background
The stevioside is a non-caloric natural sweetener extracted from Stevia rebaudiana (Stevia) of Compositae, and has sweetness 200-450 times that of sucrose. The sweet component of stevia rebaudiana Bertoni, steviol glycosides, have a common steviol skeleton, each steviol glycoside having a unique taste and sweetness depending on the number and arrangement of glucose units in the R1 and R2 positions (the structural formula of steviol glycoside is shown in FIG. 1, wherein R1 is the C19 position, and R2 is the C13 position). Among the stevioside compounds, rebaudioside a has been widely used as a sweetener in food and beverage products. But has the disadvantage of having a natural aftertaste or grassy smell, which also limits its further sugar substitution. Rebaudioside b is also one of the sweet components in steviol glycosides. The structure of the compound is similar to that of rebaudioside a, only one glucose group is different from C19 of aglycone, the sweetness is equivalent to that of rebaudioside a, no aftertaste exists, and the mouthfeel is better than that of rebaudioside a. However, the leaves of stevia rebaudiana have very low rebaudioside b content, so that the obtaining from nature is not only costly but also yield is not satisfactory. Therefore, there is a need to find a low cost method with high rebaudioside b yield and purity.
Disclosure of Invention
The invention aims to provide a method for preparing high-purity rebaudioside b by basic hydrolysis of rebaudioside a, which has low cost and high rebaudioside b yield and purity.
According to the invention, the purpose of preparing rebaudioside b from rebaudioside a by an alkali hydrolysis method is achieved by selectively hydrolyzing the glycosidic bond of C19 in rebaudioside a without destroying the glycosidic bond of C13, and the converted rebaudioside b is easy to separate out, and the subsequent separation and purification are simple and efficient.
The above object of the present invention is achieved by the following technical solutions: a method for preparing high-purity rebaudioside b by alkaline hydrolysis of rebaudioside a, comprising the steps of: selecting rebaudioside a, adding water to obtain a rebaudioside a aqueous solution, adding a base into the rebaudioside a aqueous solution to adjust the pH value to 9-12, reacting at a constant temperature of 60-100 ℃ for 2-24 hours, adding an acid to neutralize after the completion of the alkaline hydrolysis reaction, adjusting the pH value to 6-8, standing the neutralized reaction liquid, separating out a rebaudioside b crude product, and performing solid-liquid separation and drying on the rebaudioside b crude product to prepare rebaudioside b.
Preferably, the purity (mass percentage) of the rebaudioside a is 40-95%, and the concentration of the rebaudioside a aqueous solution is 50-200 mg/mL.
Preferably, the base is sodium hydroxide, potassium hydroxide, ammonia or calcium hydroxide.
Preferably, the acid is hydrochloric acid, phosphoric acid, nitric acid or sulfuric acid.
Preferably, the neutralized reaction solution is kept stand at 4-20 ℃ to separate out a crude rebaudioside b product.
Preferably, a vacuum filtration method is used for solid-liquid separation.
Preferably, the drying is carried out by air flow.
Preferably, the yield of the rebaudioside b is 65-95% (mass percentage content), and the purity is 85-97% (mass percentage content).
As a preferred embodiment of the present invention, the present invention provides a method for preparing high purity rebaudioside b by basic hydrolysis of rebaudioside a, comprising the steps of:
(1) alkali hydrolysis: selecting rebaudioside-A, adding water to prepare 50-200 mg/mL rebaudioside-A aqueous solution, adding alkali to adjust the pH value to 9-12, and reacting at a constant temperature of 60-100 ℃ for 2-24 hours;
(2) and (3) neutralization reaction: after the reaction is finished, adding common acid for neutralization, and adjusting the pH to 6-8;
(3) low-temperature crystallization: standing the neutralized reaction solution at 4-20 ℃ to separate out rebaudioside b;
(4) solid-liquid separation: separating the solid rebaudioside b by a reduced pressure suction filtration method;
(5) airflow drying: and (4) carrying out airflow drying to obtain a rebaudioside b product.
Preferably, the purity of rebaudioside a in step (1) is in the range of 40% to 95% (by mass).
Preferably, the base added in step (1) includes, but is not limited to, one of sodium hydroxide, potassium hydroxide, aqueous ammonia and calcium hydroxide.
Preferably, the acid in step (2) is hydrochloric acid, phosphoric acid, nitric acid or sulfuric acid.
Preferably, the crystallization temperature of rebaudioside b in step (3) is 4-20 ℃.
Preferably, the yield of rebaudioside b in step (5) is 65-95% (mass percentage content), and the purity is 85-97% (mass percentage content).
The invention has the following beneficial effects: the invention provides a method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis, which is simple and efficient, has high selectivity and yield, is simple in product separation and purification, has high purity of prepared products, and is suitable for mass production.
Drawings
FIG. 1 is a prior art structural formula of steviol glycosides, wherein R1 is at position C19 and R2 is at position C13;
FIG. 2 is a liquid chromatogram of a rebaudioside a control in example 1;
FIG. 3 is a liquid chromatogram of a rebaudioside b control in example 1;
FIG. 4 is a liquid chromatogram of the sample of example 1.
Detailed Description
The technical solutions of the present invention are described in detail below with reference to specific examples so that those skilled in the art can better understand and implement the technical solutions of the present invention. Reagents or materials used in the examples were commercially available, unless otherwise specified.
Example 1
Weighing 50g of rebaudioside-a with the purity of 95% (mass percentage content), adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 95 ℃ for constant-temperature reaction for 4 hours, adding hydrochloric acid for adjusting the pH value to 7.0, standing at the temperature of 20 ℃ for 16 hours, separating out a large amount of white precipitate, filtering (reduced pressure suction filtration) for washing, drying, and detecting by a liquid phase.
The liquid chromatogram of the rebaudioside a control is shown in fig. 2, the liquid chromatogram of the rebaudioside b control is shown in fig. 3, and the liquid chromatogram of the sample is shown in fig. 4.
Qualitative and quantitative analysis is carried out on a reference substance of rebaudioside a and rebaudioside b and a rebaudioside b sample through HPLC (detection method refers to stevioside national standard), retention time of the reference substance and the sample as well as a mixed sample of the reference substance and the sample is consistent, physicochemical properties of powder of the product and the reference substance are basically consistent, the product is basically judged to be rebaudioside b, meanwhile, the purity of the rebaudioside b product is obtained through comparison calculation of a liquid phase result and the reference substance, and a calculation method of yield is simplified as follows: yield% (% by mass of rebaudioside b product/(mass of raw material × purity of rebaudioside a) × 100%.
Rebaudioside b was detected in a yield of 89.4% and a purity of 95.2%.
Through the analysis of the structures of the two monomers of rebaudioside a and b and the combination of the experimental results of alkaline hydrolysis, the two monomers have a glucose group difference at the C19 position, ra (rebaudioside a) breaks a glycosidic bond under alkaline conditions to obtain rb (rebaudioside b), and after the reaction, the main product is rb (rebaudioside b), no other side products are detected, and the glycosidic bond at the C13 is judged to be not hydrolyzed basically.
Therefore, the method selectively hydrolyzes the glycosidic bond of C19 in rebaudioside a without destroying the glycosidic bond of C13, so that the rebaudioside b is prepared by the rebaudioside a through a basic hydrolysis method, the converted rebaudioside b is easy to separate out, and the subsequent separation and purification are simple and efficient.
Example 2
Weighing 50g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 60 ℃, reacting at a constant temperature for 4 hours, adding hydrochloric acid for adjusting the pH value to 7.0, standing at the temperature of 20 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase, wherein the yield of rebaudioside-b is 76.3%, and the purity of rebaudioside-b is 90.1%.
Example 3
Weighing 50g of rebaudioside-a with the purity of 50%, adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 95 ℃, reacting at a constant temperature for 4 hours, adding hydrochloric acid for adjusting the pH value to 7.0, standing at the temperature of 4 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase, wherein the yield of rebaudioside-b is 83.3%, and the purity of rebaudioside-b is 85.6%.
Example 4
Weighing 100g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 95 ℃ for constant-temperature reaction for 4 hours, adding hydrochloric acid for adjusting the pH value to 7.0, standing at the temperature of 4 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase. The yield of rebaudioside b was 87.3% and the purity was 96.6%.
Example 5
Weighing 50g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 9, stirring at the temperature of 95 ℃, reacting at a constant temperature for 4 hours, adding hydrochloric acid for adjusting the pH value to 7.0, standing at the temperature of 4 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase, wherein the yield of rebaudioside-b is 80.3%, and the purity of rebaudioside-b is 91.8%.
Example 6
Weighing 50g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 95 ℃ for constant-temperature reaction for 4 hours, adding hydrochloric acid for adjusting the pH value to 8.0, standing at the temperature of 4 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase. The yield of rebaudioside b was 74.3% and the purity was 91.1%.
Example 7
Weighing 50g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding a saturated sodium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 95 ℃, reacting at a constant temperature for 6 hours, adding hydrochloric acid for adjusting the pH value to 7.0, standing at the temperature of 20 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase, wherein the yield of rebaudioside-b is 69.9%, and the purity of rebaudioside-b is 92.1%.
Example 8
Weighing 50g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding ammonia water for adjusting the pH value to 12, stirring at the temperature of 95 ℃ for constant-temperature reaction for 6 hours, adding phosphoric acid for adjusting the pH value to 8.0, standing at the temperature of 4 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase. The yield of rebaudioside b was 65.3% and the purity was 89.6%.
Example 9
Weighing 50g of rebaudioside-a with the purity of 95%, adding 1L of water for dissolving, adding a potassium hydroxide solution for adjusting the pH value to 12, stirring at the temperature of 95 ℃ for constant-temperature reaction for 4 hours, adding sulfuric acid for adjusting the pH value to 8.0, standing at the temperature of 4 ℃ for 16 hours, separating out a large amount of white precipitate, filtering, washing with water, drying, and detecting by a liquid phase. The yield of rebaudioside b was 77.7% with a purity of 90.6%.
The above embodiments are only used for illustrating the present invention, and the scope of the present invention is not limited to the above embodiments. The object of the present invention can be achieved by those skilled in the art based on the above disclosure, and any improvements and modifications based on the concept of the present invention fall within the protection scope of the present invention, which is defined by the claims.
Claims (8)
1. A method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis is characterized by comprising the following steps: selecting rebaudioside a, adding water to obtain a rebaudioside a aqueous solution, adding a base into the rebaudioside a aqueous solution to adjust the pH value to 9-12, reacting at a constant temperature of 60-100 ℃ for 2-24 hours, adding an acid to neutralize after the completion of the alkaline hydrolysis reaction, adjusting the pH value to 6-8, standing the neutralized reaction liquid, separating out a rebaudioside b crude product, and performing solid-liquid separation and drying on the rebaudioside b crude product to prepare rebaudioside b.
2. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: the purity of the rebaudioside a is 40% -95%, and the concentration of the rebaudioside a aqueous solution is 50-200 mg/mL.
3. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: the alkali is sodium hydroxide, potassium hydroxide, ammonia water or calcium hydroxide.
4. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: the acid is hydrochloric acid, phosphoric acid, nitric acid or sulfuric acid.
5. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: and standing the neutralized reaction liquid at 4-20 ℃ to precipitate a rebaudioside b crude product.
6. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: and a decompression suction filtration method is adopted during solid-liquid separation.
7. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: and when drying, airflow drying is adopted.
8. The method for preparing rebaudioside b by alkaline hydrolysis of rebaudioside a according to claim 1, wherein: the yield of rebaudioside b is 65-95%, and the purity is 85-97%.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210275100.XA CN114805456A (en) | 2022-03-21 | 2022-03-21 | Method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210275100.XA CN114805456A (en) | 2022-03-21 | 2022-03-21 | Method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114805456A true CN114805456A (en) | 2022-07-29 |
Family
ID=82530613
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210275100.XA Pending CN114805456A (en) | 2022-03-21 | 2022-03-21 | Method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114805456A (en) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102796790A (en) * | 2012-08-14 | 2012-11-28 | 成都南诺格生物科技有限责任公司 | Method for conversing steviolbioside to rebaudiodside B |
CN104725437A (en) * | 2013-12-19 | 2015-06-24 | 江南大学 | Method for preparation of laminaribiose and rebaudiodside B by basic hydrolysis of rebaudiodside I |
CN112322686A (en) * | 2020-11-27 | 2021-02-05 | 四川盈嘉合生科技有限公司 | Method for producing rebaudioside B by enzyme method |
CN113583064A (en) * | 2021-08-28 | 2021-11-02 | 诸城市浩天药业有限公司 | Process method for producing rebaudioside B by high-temperature cracking method |
-
2022
- 2022-03-21 CN CN202210275100.XA patent/CN114805456A/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102796790A (en) * | 2012-08-14 | 2012-11-28 | 成都南诺格生物科技有限责任公司 | Method for conversing steviolbioside to rebaudiodside B |
CN104725437A (en) * | 2013-12-19 | 2015-06-24 | 江南大学 | Method for preparation of laminaribiose and rebaudiodside B by basic hydrolysis of rebaudiodside I |
CN112322686A (en) * | 2020-11-27 | 2021-02-05 | 四川盈嘉合生科技有限公司 | Method for producing rebaudioside B by enzyme method |
CN113583064A (en) * | 2021-08-28 | 2021-11-02 | 诸城市浩天药业有限公司 | Process method for producing rebaudioside B by high-temperature cracking method |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107532189B (en) | Method for the enzymatic modification of steviol glycosides, modified steviol glycosides obtainable thereby and their use as sweeteners | |
AU2015219212B2 (en) | Process for separation, isolation and characterization of steviol glycosides | |
CN101472487B (en) | Rebaudioside a composition and method for purifying rebaudioside a | |
CN113683712B (en) | Steviol glycosides | |
JP2020505907A (en) | Food ingredients from Stevia rebaudiana | |
US11059843B2 (en) | Stevioside M crystal form, preparation method therefor and use thereof | |
CA2857091A1 (en) | Processes of purifying steviol glycosides reb c | |
US3429873A (en) | Preparation of hesperetin dihydrochalcone glucoside | |
EP3708672B1 (en) | Method for producing transglucosylated steviol glycoside using lactobacillus mali | |
CN104478706A (en) | Method for preparing steviol | |
CN114805456A (en) | Method for preparing high-purity rebaudioside b by adopting rebaudioside a alkaline hydrolysis | |
KR101199821B1 (en) | Method for preparing functional natural high intensity sweetener stevio- oligoglycosides by using glycosyltransferases | |
CN105906674B (en) | A method of the separating high-purity Rc from steviol glycoside refinement mother liquor | |
CN112300231B (en) | Method for extracting high-purity stevioside | |
CN113024627B (en) | Dammarane type triterpenoid saponin compound and preparation method and application thereof | |
CN114031655A (en) | Crystallization method of stevioside | |
KR20190002364A (en) | Synthesis method of rubusoside-fructoside using levansucrase | |
CN105037455B (en) | The synthetic method of alkyl β D glucopyranosides | |
CN113201034A (en) | Obtaining high-purity stevioside from primary crystallization mother liquor of stevioside through secondary crystallization and enriching rebaudioside C | |
CN108707163A (en) | A kind of preparation method of steviol glucoside member | |
CN113527378B (en) | Method for recovering stevioside from industrial stevioside mother liquor sugar through crystallization | |
CN110143942B (en) | Method for preparing icaritin and rhamnose syrup by organic acid hydrolysis of icariin | |
CN111850071A (en) | High-purity enzyme-modified stevioside and preparation method thereof | |
JPH0612977B2 (en) | Method for producing fructose condensate | |
Kusakabe et al. | An easy method for the preparation of sophorose from stevioside |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |