A method for extracting and purifying bosogenin

Abstract

The present invention discloses a method for extracting and purifying boson, and belongs to the technical field of enzyme-catalyzed boson production. The method comprises the following steps: (1) inactivating the enzyme-catalyzed reaction liquid containing boson, filtering it through a ceramic membrane, and obtaining a filtrate; (2) desalting the filtrate through electrodialysis to obtain a desalted liquid; (3) decolorizing and removing impurities from the desalted liquid through a zeolite molecular sieve column and an alumina column to obtain a decolorized liquid; (4) concentrating and freeze-drying the decolorized liquid to obtain boson. The method of the present invention has a simple process, can effectively remove impurities such as proteins, pigments, and salts brought about in the enzyme reaction process, abandons the use of toxic organic reagents, is low in cost, is green and environmentally friendly, has high product purity, and has good output quality, and is suitable for industrial production.

Description

Technical Field

[0001] The invention belongs to the technical field of enzyme-catalyzed production of bosera, and in particular relates to a method for extracting and purifying bosera. Background Art

[0002] Bosin, chemically known as hydroxypropyl tetrahydropyrantriol, is a xylose derivative with anti-aging active ingredients. Bosin can not only stimulate the production of aminoglucosan (GAGs), the main component of the extracellular matrix of the skin, effectively preventing the loss of skin moisture; it can also promote the synthesis of skin collagen, enhance the connectivity between the dermis and the epidermis, and improve the bonding ability between the dermis and the epidermis. Bosin, as a cosmetic raw material, has moisturizing, repairing, anti-wrinkle and anti-aging effects.

[0003] However, the bosogen extracted from the enzyme-catalyzed reaction solution in the current process has low purity, contains more impurities, and is complicated to operate. In addition, most of the extractions are done with organic reagents, which is costly and causes serious environmental pollution. Therefore, it is very important to provide a method for extracting high-purity bosogen for the industrial application of bosogen. Summary of the invention

[0004] In view of the deficiencies in the prior art, the present invention aims to provide a method for extracting and purifying bosogenin, which has simple process operation, low cost and is suitable for industrial production.

[0005] In order to solve the above technical problems, the technical solutions provided by the present invention are as follows:

[0006] A method for extracting and purifying bosogenin comprises the following steps:

[0007] (1) inactivating the enzyme-catalyzed reaction solution containing bosine, filtering it through a ceramic membrane, and obtaining a filtrate;

[0008] (2) Desalting the filtrate by electrodialysis to obtain a desalted solution;

[0009] (3) Decolorizing and removing impurities from the desalted liquid by passing it through a zeolite molecular sieve column and an alumina column to obtain a decolorized liquid;

[0010] (4) Concentrate the decolorized solution and freeze-dry it to obtain boson.

[0011] Furthermore, in step (1), the pore size of the ceramic membrane is 50-200 nm, preferably 50 nm. The reaction solution is filtered through the ceramic membrane to remove bacteria and proteins.

[0012] Furthermore, in step (1), at a wavelength of 450 nm, the transmittance of the filtrate is greater than 94%.

[0013] Furthermore, in step (2), the electrodialysis uses a heterogeneous ion exchange membrane.

[0014] Furthermore, in step (2), the conductivity of the desalted solution is less than 100 μS / cm.

[0015] Furthermore, in step (3), the zeolite molecular sieve is one or more of ZSM-5 molecular sieve, MCM-22 molecular sieve or 13X molecular sieve; the volume ratio of zeolite molecular sieve to alumina is 1:1-5, preferably 3:7. Zeolite molecular sieve can decolorize and remove impurities from the desalted liquid, and alumina can further decolorize the desalted liquid to ensure the purity of the finished boson, and both can be reused.

[0016] Furthermore, in step (3), after decolorization and impurity removal by zeolite molecular sieve column and alumina column, the decolorization rate is 90-99%.

[0017] Furthermore, in step (4), the concentration is performed by vacuum concentration, the concentration temperature is 60-80° C., and the concentration vacuum degree is 0.07-0.08 MPa.

[0018] Furthermore, in step (4), after the decolorizing solution is concentrated, the concentration of boron sulfoxide in the concentrated solution is 900-1000 g / L.

[0019] Furthermore, in step (4), the purity of the boson is greater than 99.5%.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) The present invention adopts a membrane method to remove bacteria and proteins, and then uses zeolite molecular sieve and alumina to decolorize and remove impurities to prepare bosogen. The purity of bosogen is greater than 99.5%, and the product purity is high.

[0022] (2) The zeolite molecular sieve and alumina selected in the present invention can be reused many times, which can not only reduce the use of raw materials in the purification process, reduce production costs, and increase economic benefits, but also reduce waste emissions and reduce environmental pollution, which is green and environmentally friendly.

[0023] (3) The method of the present invention is simple to operate and can effectively remove impurities such as protein, pigment, salt, etc. brought about by the enzyme reaction process, and abandons the use of toxic organic reagents. It is low-cost, green and environmentally friendly, and suitable for industrial production. DETAILED DESCRIPTION

[0024] Example 1

[0025] (1) Inactivate 50 L of enzyme-catalyzed reaction solution containing bosine, filter it with a ceramic membrane with a pore size of 100 nm, and obtain a filtrate with a transmittance of 95.5% and a conductivity of 15 mS / cm at 450 nm;

[0026] (2) The filtrate is subjected to electrodialysis desalination using a heterogeneous ion exchange membrane to obtain a desalted liquid with an electrical conductivity of 82 μS / cm;

[0027] (3) Zeolite molecular sieve ZSM-5 and alumina with a volume ratio of 1:1 were loaded into columns respectively, and then the desalted liquid was passed through the zeolite molecular sieve ZSM-5 column and the alumina column connected in series in sequence to remove impurities and decolorize to obtain a decolorized liquid with a decolorization rate of 97%;

[0028] (4) The decolorized liquid was concentrated under vacuum conditions at a concentration temperature of 60°C and a vacuum degree of 0.07 MPa to obtain a concentrated liquid with a concentration of 950 g / L; the concentrated liquid was then freeze-dried to obtain boson. The purity of boson was found to be 99.5%.

[0029] Example 2

[0030] (1) Inactivate 100 L of enzyme-catalyzed reaction solution containing bosine, filter it with a ceramic membrane with a pore size of 50 nm, and obtain a filtrate with a transmittance of 96% and a conductivity of 15 mS / cm at 450 nm;

[0031] (2) The filtrate is subjected to electrodialysis desalination using a heterogeneous ion exchange membrane to obtain a desalted liquid with a conductivity of 70 μS / cm;

[0032] (3) Zeolite molecular sieve 13X and alumina with a volume ratio of 3:7 were loaded into columns respectively, and then the desalted liquid was passed through the zeolite molecular sieve 13X column and the alumina column connected in series in sequence to remove impurities and decolorize to obtain a decolorized liquid with a decolorization rate of 99%;

[0033] (4) The decolorized liquid was concentrated under vacuum conditions at a concentration temperature of 70°C and a vacuum degree of 0.07 MPa to obtain a concentrated liquid with a concentration of 980 g / L; the concentrated liquid was then freeze-dried to obtain boson. The purity of boson was found to be 99.8%.

[0034] Example 3

[0035] (1) Inactivate 200 L of enzyme-catalyzed reaction solution containing bosine, filter it with a ceramic membrane with a pore size of 200 nm, and obtain a filtrate with a transmittance of 94.3% and a conductivity of 15 mS / cm at 450 nm;

[0036] (2) The filtrate is subjected to electrodialysis desalination using a heterogeneous ion exchange membrane to obtain a desalted liquid with a conductivity of 72 μS / cm;

[0037] (3) Zeolite molecular sieve MCM-22 and alumina with a volume ratio of 2:9 were loaded into columns and pretreated, and then the desalted liquid was passed through the zeolite molecular sieve MCM-22 column and alumina column in series to remove impurities and decolorize, with a decolorization rate of 99%;

[0038] (4) The decolorized liquid was concentrated under vacuum conditions at a concentration temperature of 80°C and a vacuum degree of 0.08 MPa to obtain a concentrated liquid with a concentration of 1000 g / L; the concentrated liquid was then freeze-dried to obtain boson. The purity of boson was found to be 99.6%.

[0039] Comparative Example 1

[0040] The reaction steps are the same as those in Example 2, except that zeolite molecular sieve is not used for impurity removal and decolorization, specifically:

[0041] (3) Alumina is loaded on the column, and the desalted liquid is passed through the alumina column to remove impurities and decolorize to obtain a decolorized liquid with a decolorization rate of 86%.

[0042] The purity of boson was tested to be 89% and the sample was light yellow in appearance.

[0043] Comparative Example 2

[0044] The reaction steps are the same as those in Example 2, except that aluminum oxide is not used for decolorization. Specifically,

[0045] (3) Zeolite molecular sieve 13X was loaded into a column, and the desalted liquid was passed through the zeolite molecular sieve 13X column to remove impurities and decolorize. The decolorization rate was 82%.

[0046] The purity of boson was tested to be 92% and the sample was yellow in appearance.

[0047] Comparative Example 3

[0048] The reaction steps are the same as those of Example 2, except that step (3) is not performed.

[0049] The purity of boson was tested to be 85% and the sample was light brown in appearance.

[0050] Experimental Example 1

[0051] The purity of boson and the appearance of the samples in Example 2 and Comparative Examples 1-3 are listed in Table 1.

[0052] Table 1. Purity of boson and sample appearance

[0053]

[0054] As can be seen from Table 1, the purity of Example 2 Bose is 99.8%, the purity of Comparative Example 1 Bose is 89%, the purity of Comparative Example 2 Bose is 92%, and the purity of Comparative Example 3 Bose is 85%, indicating that the purity of Bose prepared by the method of the present invention is high. It can also be seen from the appearance of the samples that the samples that have not been decolorized by zeolite molecular sieve columns and alumina columns have a light brown appearance, while the samples that have been decolorized by zeolite molecular sieve columns and alumina columns have a white appearance. It can be seen that the method of the present invention uses membrane sterilization and protein removal, and then zeolite molecular sieves and alumina can be used to decolorize and remove impurities to obtain high-purity Bose.

[0055] Experimental Example 2

[0056] (2-1) One-time reuse of zeolite molecular sieve 13X column and alumina column:

[0057] (1) Inactivate 100 L of enzyme-catalyzed reaction solution containing bosine, filter it with a ceramic membrane with a pore size of 50 nm, and obtain a filtrate with a transmittance of 96% and a conductivity of 15 mS / cm at 450 nm;

[0058] (2) The filtrate is subjected to electrodialysis desalination using a heterogeneous ion exchange membrane to obtain a desalted liquid with a conductivity of 70 μS / cm;

[0059] (3) The zeolite molecular sieve column and alumina column after decolorization and impurity removal in Example 2 were washed with 5 times the column volume of 95% ethanol and washed with purified water until there was no ethanol smell. The alumina column was soaked in 3% hydrochloric acid solution for 24 hours and then washed with water until neutral, which were recorded as zeolite molecular sieve 13X column 1 and alumina column 1. The desalted liquid was then passed through the zeolite molecular sieve 13X column 1 and alumina column 1 connected in series in sequence to remove impurities and decolorize to obtain a decolorized liquid with a decolorization rate of 98%;

[0060] (4) The decolorized liquid was concentrated under vacuum conditions at a concentration temperature of 70°C and a vacuum degree of 0.07 MPa to obtain a concentrated liquid with a concentration of 980 g / L; the concentrated liquid was then freeze-dried to obtain boson. The purity of boson was found to be 99.7%.

[0061] (2-2) Five times of repeated use of zeolite molecular sieve 13X column and alumina column:

[0062] The experiment was carried out according to the above operation, and the zeolite molecular sieve 13X column and the alumina column were reused five times:

[0063] (1) Inactivate 100 L of enzyme-catalyzed reaction solution containing bosine, filter it with a ceramic membrane with a pore size of 50 nm, and obtain a filtrate with a transmittance of 96% and a conductivity of 15 mS / cm at 450 nm;

[0064] (2) The filtrate is subjected to electrodialysis desalination using a heterogeneous ion exchange membrane to obtain a desalted liquid with an electrical conductivity of 70 μS / cm;

[0065] (3) The zeolite molecular sieve column and alumina column after decolorization and impurity removal in Example 2 were washed with 5 times the column volume of 95% ethanol and washed with purified water until there was no ethanol smell. The alumina column was soaked in 3% hydrochloric acid solution for 24 hours and then washed with water until neutral, which were recorded as zeolite molecular sieve 13X column 5 and alumina column 5. The desalted liquid was then passed through the zeolite molecular sieve 13X column 5 and alumina column 5 connected in series in sequence to remove impurities and decolorize to obtain a decolorized liquid with a decolorization rate of 96%;

[0066] (4) The decolorized liquid was concentrated under vacuum conditions at a concentration temperature of 70°C and a vacuum degree of 0.07 MPa to obtain a concentrated liquid with a concentration of 980 g / L; the concentrated liquid was then freeze-dried to obtain boson. The purity of boson was found to be 99.5%.

[0067] The decolorization rates of Example 2 and Experimental Example 2 and the purity of phosphine are listed in Table 2.

[0068] Table 2. Decolorization rate and purity of phosphodiesterone

[0069]

[0070] As can be seen from Table 2, the decolorization rate of Example 2 is 99%, and the purity of Bose is 99.8%. In Experimental Example (2-1), the zeolite molecular sieve column and the alumina column are reused after regeneration treatment, and the decolorization rate is 98%, and the purity of Bose is 99.7%. In Experimental Example (2-2), the zeolite molecular sieve column and the alumina column that have been reused after multiple regeneration treatments are reused again after regeneration treatment, and the decolorization rate is 96%, and the purity of Bose is 99.5%. It can be seen that the zeolite molecular sieve column and alumina column selected in the present invention can be reused many times, which can not only reduce production costs, but also reduce waste emissions, reduce environmental pollution, and be green and environmentally friendly.

Claims

1. A method for extracting and purifying bosine, characterized in that: The following steps are involved: (1) inactivating the enzyme-catalyzed reaction solution containing bosine, filtering it through a ceramic membrane, and obtaining a filtrate; (2) Desalting the filtrate by electrodialysis to obtain a desalted solution; (3) Decolorizing and removing impurities from the desalted liquid by passing it through a zeolite molecular sieve column and an alumina column to obtain a decolorized liquid; (4) Concentrate the decolorized solution and freeze-dry it to obtain boson.

2. The method for extracting and purifying bosogen according to claim 1, characterized in that: In step (1), the pore size of the ceramic membrane is 50-200 nm.

3. The method for extracting and purifying bosogen according to claim 1, characterized in that: In step (1), at a wavelength of 450 nm, the transmittance of the filtrate is greater than 94%.

4. The method for extracting and purifying bosogenin according to any one of claims 1 to 3, characterized in that: In step (2), the electrodialysis uses a heterogeneous ion exchange membrane.

5. The method for extracting and purifying bosogen according to claim 4, characterized in that: In step (2), the conductivity of the desalted solution is less than 100 μS / cm.

6. The method for extracting and purifying bosogen according to claim 5, characterized in that: In step (3), the zeolite molecular sieve is one or more of ZSM-5 molecular sieve, MCM-22 molecular sieve or 13X molecular sieve; the volume ratio of zeolite molecular sieve to alumina is 1:1-5.

7. The method for extracting and purifying bosogen according to claim 1, characterized in that: In step (4), the concentration is performed by vacuum concentration, the concentration temperature is 60-80° C., and the concentration vacuum degree is 0.07-0.08 MPa.

8. The method for extracting and purifying bosogenin according to claim 7, characterized in that: In step (4), after the decolorizing solution is concentrated, the concentration of benzophenone in the concentrated solution is 900-1000 g / L.

9. The method for extracting and purifying bosogenin according to claim 8, characterized in that: In step (4), the purity of the boson is greater than 99.5%.

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