CN109796544B - Industrial preparation method and inspection method of 2, 6-di-O-methyl-beta-cyclodextrin - Google Patents
Industrial preparation method and inspection method of 2, 6-di-O-methyl-beta-cyclodextrin Download PDFInfo
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Abstract
The invention relates to an industrial preparation method and a detection method of 2, 6-di-O-methyl-beta-cyclodextrin, which comprises the following steps: s1: drying the beta-cyclic dextrin before feeding; s2: mixing anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide under low temperature, and stirring; then adding the dried beta-cyclodextrin and stirring, then dropwise adding methyl sulfate and stirring, and continuing stirring for a preset time after all dropwise adding is finished; s3: closing the low-temperature system in the S2, continuously stirring and heating to 20-30 ℃, keeping the temperature and continuously reacting for 20-60 h; s4: extracting the product of S3 with chloroform and neutralizing; s5: sequentially carrying out salt washing and water washing on the neutralized product, carrying out suction filtration and collecting filtrate; s6: dehydrating the filtrate, and then distilling and concentrating the filtrate until the filtrate is concentrated to be viscous; s7: and drying the concentrated solution, and then recrystallizing to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Description
Technical Field
The invention relates to the technical field of synthesis of organic high molecular compounds, in particular to an industrial preparation method and a detection method of 2, 6-di-O-methyl-beta-cyclodextrin.
Background
[2, 6-di-O-methyl group]7-beta-cyclodextrin, Heptakis (2,6-di-O-methyl) -beta-cyclodextrin is the standard name of the product, and can be abbreviated as 2, 6-di-O-methyl-beta-cyclodextrin or 2, 6-di-O-methyl-beta-cyclodextrin, and part of foreign language information is abbreviated as 2, 6-di-O-methyl-beta-cyclodextrin with the molecular formula of C56H98O35Molecular weight 1331.36, white powder in appearance, the chemical formula is as follows:
2, 6-di-O-methyl-beta-cyclodextrin is a product obtained by performing directional methylation reaction on beta-cyclodextrin serving as a main raw material. Beta-cyclodextrin (beta-cyclodextrin), abbreviated as beta-CD, is a white solid formed by 7D-glucopyranoses in a chair-like configuration, condensed and cyclized by alpha (1-4) glycosidic bond linkage, and has hydroxyl groups at three positions of 2,3 and 6 of each glucopyranose unit in the structural formula. Among them, cyclodextrin is a group of cyclic oligosaccharides formed by alpha-1, 4-glycosidic linkages and produced by fermentation of starch with glucose transferase, and is widely studied since then, it is classified into various kinds according to the difference in the number of glucose units in the molecule, and alpha, beta, gamma-cyclodextrin (alpha, beta, gamma-CD) is the most common 3 kinds, which in turn contain 6, 7, and 8 glucose units. Beta-cyclodextrin (beta-CD) is a compound which is formed by connecting 7D-glucopyranose units end to end and has a cone-shaped structure, wherein the inner part is hydrophobic, the outer part is hydrophilic, the 6-position primary hydroxyl is positioned at the small opening end, and the 2-position secondary hydroxyl is positioned at the large opening end. The special cavity and the internal hydrophobic environment can form an inclusion compound with object molecules or groups with matched sizes, polarities and properties, and have great influence on the aspects of improving the solubility of the object molecules, improving the stability of the object molecules, releasing slowly and the like, so that the cyclodextrin is widely applied to the aspects of medicines, environments, materials, enzyme catalysis, daily chemical industry, packaging industry, food industry and the like.
2, 6-di-O-methyl-beta-cyclodextrin is mainly used as a growth promoter of a protective antigen of pertussis bacteria, and is also used for application and theoretical research in the fields of biochemistry and chemistry. The production promoter of the pertussis bacterium protective antigen can eliminate the side effect of vaccine, and the pertussis bacterium serving as a vaccine which is required to be injected for the first time in the newly born children is cultured and grown by a stationary method at home and abroad for a long time. The bacterin cultured by the method has low yield, and various side effects such as fever, crazy suction, suppuration and the like can be caused after the bacterin cultured and grown is injected on a human body. The 2, 6-di-O-methyl-beta-cyclodextrin is used as a pertussis protective antigen growth promoter, the yield of the pertussis protective antigen is improved by 2-3 times compared with that of a static method when the pertussis protective antigen is cultured under the condition of a cradle, and the vaccine cultured and grown by the method has no side effect after being injected into a human body. The world health organization has approved the use of 2, 6-di-O-methyl-beta-cyclodextrin as promoter to culture and grow pertussis bacteria protective antigen vaccine. The pertussis protective antigen bacterins cultured by using 2, 6-di-O-methyl-beta-cyclodextrin developed by people as an accelerant by several domestic biological product research institutes pass the standards of the Ministry of health, are applied in large batches for many years, have stable performance and meet the requirement of vaccine culture.
With respect to [2, 6-di-O-methyl]7Many studies have been made on the preparation method of-beta-cyclodextrin. For example: studies on the synthesis of 2,6-di-O-methyl- β -cyclodextrin are published by Chiyan et al in publications (Zhejiang medical university journal 1997, 26, 2). This article describes the laboratory preparation of 2,6-di-O-methyl- β 0-cyclodextrin. The method can be prepared in a small dose in a laboratory, and industrial mass production cannot be realized. A study on the synthesis, toxicity and solubilization of partially methylated beta-cyclodextrin was published in the publication by Song quan et al (journal of Chinese medical industry 1999, 11, 008). This article describes the laboratory preparation of methyl- β -cyclodextrin. The method can be prepared in a small dose in a laboratory, and industrial mass production cannot be realized. Populus et al published a study on the preparation and characterization of partially methylated beta-cyclodextrin mixtures in publications (journal of chinese medical industry 2004, 35, 5). This article describes the laboratory preparation of methyl- β -cyclodextrin. The method can be prepared in a small dose in a laboratory, and industrial mass production cannot be realized. A study on a method for detecting the content of iodoethersalicylamide in animal-derived foods and a study on a process for synthesizing 2,6-di-O-methyl- β -cyclodextrin are disclosed in publications (Shenyang pharmaceutical university Master's academic paper 2008, 5) by Yanfen et al. This article describes the laboratory preparation of 2,6-di-O-methyl- β -cyclodextrin. The method can be prepared in a small dose in a laboratory, and industrial mass production cannot be realized. A study on a novel synthesis process of methylated- β -cyclodextrin was published in published publications (journal of chemical engineering 2009, 12) of university, by ganyongjiang et al. This article describes a laboratory preparation of 2,6-di-O-methyl- β -cyclodextrin using dimethyl carbonate instead of dimethyl sulfate. The method can be prepared in small dose in a laboratory, cannot realize industrialized mass production,and whether the industrial production yield is reduced by replacing dimethyl sulfate with dimethyl carbonate is still to be verified. An article on the progress of the research on the use of beta-cyclodextrin and its derivatives was published by constantan et al in the publication (proceedings of the university of salted yang 2013.3). The research article theoretically elaborates the selective modification method of beta-cyclodextrin, including the 2 nd and 6 th reaction principles and the difficulty degree. The research content of the article is in a theoretical research stage, and industrial mass production cannot be realized.
In addition, the 2,6-di-O-methyl is not found by referring to relevant inspection standard data at home and abroad]7-assay standard for β -cyclodextrin. Further, the catalogues of several large reagent companies such as Sigam Chemical Company, FluKa Chemika-BichemiKa Company, Aldrich Chemical Company. inc in China were referred to, and these larger foreign manufacturers did not give the same specification index to the product. In addition to the molecular formula and molecular weight of the compound, some companies give nuclear magnetic resonance spectrum, carbon element analysis table, Raman spectrum or mass spectrum data and the like of the product; the indexes for controlling the product quality are less.
Based on the method, the industrial mass preparation method and the inspection method of the novel 2, 6-di-O-methyl-beta-cyclodextrin are of great significance.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide an industrial preparation method and a detection method of 2, 6-di-O-methyl-beta-cyclodextrin. The industrial preparation method of the 2, 6-di-O-methyl-beta-cyclodextrin provided by the invention is initiated at home, breaks through the limitation that the original production can be carried out only in a small amount in a laboratory, successfully solves the current situation that the 2, 6-di-O-methyl-beta-cyclodextrin cannot be industrially prepared in a large amount at home, further has a good promotion effect on the mass production of pertussis vaccines without side effects and the prevention of children pertussis, and has remarkable social benefits. The preparation method has the technical key point that the electronegativity of the three hydroxyl groups is different by using a proper reaction system, so that the nucleophilic substitution reaction is carried out by using a methylating agent with different difficulty and the 2, 6-di-O-methyl-beta-cyclodextrin is generated.
Therefore, the invention provides the following technical scheme:
in a first aspect, the present invention provides an industrial process for the preparation of 2,6-di-O-methyl- β -cyclodextrin, comprising the steps of: s1: drying the beta-cyclic dextrin before feeding until the water loss rate is 3-20%; s2: mixing anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide under low temperature, and stirring; then adding the dried beta-cyclodextrin and stirring, then dropwise adding methyl sulfate and stirring, and continuing stirring for a first preset time after all dropwise adding is finished; s3: closing the low-temperature system in the S2, continuously stirring and heating to 20-30 ℃, keeping the temperature and continuously reacting for 20-60 h; s4: extracting the product obtained in the step S3 by using trichloromethane, and then adding saturated sodium bicarbonate solution for neutralization; s5: sequentially carrying out salt washing and water washing on the neutralized product, then carrying out suction filtration and collecting filtrate; s6: dehydrating the filtrate, and then distilling and concentrating the filtrate until the filtrate is concentrated to be viscous; s7: drying the concentrated solution to remove the solvent to obtain a crude product; and recrystallizing the crude product to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Preferably, in S1: the drying treatment specifically comprises the following steps: the temperature is 90-130 ℃, and the time is 2-15 h.
Preferably, in S2: the low temperature is-10 ℃ to 10 ℃; the mass ratio of anhydrous barium oxide, barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 30: 50: (40-70): (10-35); and the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: (10-25).
Preferably, in S2: adding the beta-cyclodextrin in several times, wherein the interval between every two adjacent times is 5-30 min, and continuously stirring for 0.5-3 h after the beta-cyclodextrin is completely added; adding methyl sulfate for several times, dropwise adding each part for 1.5-4 h, and continuously stirring for 15-60 min; the first preset time is 10-25 h; more preferably, the beta-cyclodextrin and the methyl sulfate are added for 1-5 times.
Preferably, in S4: the neutralization is specifically as follows: slowly adding a saturated sodium bicarbonate solution into the extracted product at the temperature of 10-30 ℃, continuously stirring for a second preset time, standing for layering, and taking a liquid phase; wherein the saturated sodium bicarbonate solution is added for 1-5 hours, and the second preset time is 5-18 hours.
Preferably, in S5: the salt washing specifically comprises the following steps: washing the neutralized product for several times by using saturated salt water, wherein in the washing process, the temperature is 10-30 ℃, stirring is carried out for 0.5-3 h each time, and standing and layering are carried out for 0.5-3 h; the water washing specifically comprises the following steps: washing the neutralized product for several times by using pure water, wherein in the washing process, the temperature is 10-30 ℃, stirring is carried out for 0.5-3.5 h each time, and standing and layering are carried out for 0.5-3.5 h; more preferably, the salt washing and the water washing are performed in 2 to 5 times.
Preferably, in S6: the dehydration treatment specifically comprises the following steps: spreading anhydrous sodium sulfate into the filtrate, fully stirring, and standing for 5-18 h; in the distillation and concentration, the temperature is 70-150 ℃.
Preferably, in S7: the drying process comprises the following steps: the drying process comprises the following steps: the temperature is 60-135 ℃, and the time is 30-60 h; in the recrystallization process: mixing the crude product and trichloromethane according to the proportion of 1: (3-10) re-dissolving, standing for 5-15 h, performing suction filtration, distilling, concentrating, recovering the solvent, and drying the concentrated solution again.
Preferably, the step of S7 further comprises: and (3) crushing the target product 2, 6-di-O-methyl-beta-cyclodextrin to 40-70 meshes.
In a second aspect, the 2,6-di-O-methyl- β -cyclodextrin obtained is prepared according to the process of the present invention.
In a third aspect, the invention provides a method for testing 2, 6-di-O-methyl-beta-cyclodextrin, wherein in the content percentage of elements, the content percentage of carbon elements is 49.9-51.10%, and the content percentage of hydrogen elements is 7.10-7.80; in the water dissolution test, the solution is transparent and has no mechanical impurities; in the burning residue test, the residue amount is less than or equal to 0.1 percent.
The technical scheme provided by the invention has the following beneficial effects:
(1) the industrial preparation method of the 2, 6-di-O-methyl-beta-cyclodextrin provided by the invention is initiated at home, breaks through the limitation that the original production can be carried out only in a small amount in a laboratory, successfully solves the current situation that the 2, 6-di-O-methyl-beta-cyclodextrin cannot be industrially prepared in a large amount at home, further has good promotion effects on mass production of pertussis vaccines without side effects and prevention of children pertussis, and has remarkable social benefits.
(2) The yield of the preparation method provided by the invention reaches an international higher level. Compared with the international similar technology, the 2, 6-di-O-methyl-beta-cyclodextrin produced by the preparation method has the following advantages: the yield of the product obtained by various technologies is generally 70-80%, while the yield of the 2, 6-di-O-methyl-beta-cyclodextrin produced by the method is more than 80%, and reaches 85% on average, thereby reaching the higher production level of similar products abroad.
(3) The three wastes generated in the preparation method of the invention are treated and meet the national sanitary and environmental protection requirements. In addition, the invention sets up the inspection standard of 2, 6-di-O-methyl-beta-cyclodextrin, so that the performance detection of the product is more strict, scientific and accurate.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a schematic process diagram of 2, 6-di-O-methyl-beta-cyclodextrin in the example of the present invention;
FIG. 2 is a flow chart of the preparation of 2, 6-di-O-methyl-beta-cyclodextrin in the example of the present invention.
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. The following examples are only for illustrating the technical solutions of the present invention more clearly, and therefore are only examples, and the protection scope of the present invention is not limited thereby.
The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional reagent store unless otherwise specified. In the quantitative tests in the following examples, three replicates were set, and the data are the mean or the mean ± standard deviation of the three replicates.
The invention provides an industrial mass preparation method of 2, 6-di-O-methyl-beta-cyclodextrin, as shown in figures 1 and 2, comprising the following steps:
s1: drying the beta-cyclic dextrin before feeding, controlling the temperature at 90-130 ℃, the drying time at 2-15 h, and the water loss rate at 3-20%.
S2: sequentially adding anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide into a reaction kettle 1, fully stirring, and cooling to +10 ℃ to-10 ℃ for keeping; the mass ratio of anhydrous barium oxide, barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 30: 50: (40-70): (10-35), adding the beta-cyclodextrin for 1-5 times, separating every two adjacent times for 5-30 min, and continuously stirring for 0.5-3 h after all the beta-cyclodextrin are added; and then dropwise adding methyl sulfate, wherein the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: (10-25), averagely dividing methyl sulfate into 1-5 parts, dropwise adding each part for 1.5-4 h, and stirring for 15-60 min; and after all the dropwise adding is finished, continuously stirring and reacting for 10-25 h.
S3: and (5) stopping the refrigeration system in the S2, stirring continuously, heating to 20-30 ℃, and keeping the temperature to continue reacting for 20-60 hours.
S4: and (3) extracting the product obtained in the step (S3) with trichloromethane, then placing the extracted product into a reaction kettle 2, stirring, controlling the kettle temperature at 10-30 ℃, slowly adding a saturated sodium bicarbonate solution for neutralization, continuing to stir for 5-18 h after 1-5 h, standing for layering, and discharging a product solution.
S5: sequentially carrying out salt washing on the neutralized product, washing the neutralized product by adopting saturated salt water for 2-5 times, controlling the temperature of a kettle to be 10-30 ℃, stirring for 0.5-3 h each time, and standing for layering for 0.5-3 h; then washing with pure water for 2-5 times, controlling the temperature of the kettle at 10-30 ℃, stirring for 0.5-3.5 h each time, and standing for layering for 0.5-3.5 h; and then, carrying out suction filtration on the product solution after washing by using a G4 sand core funnel, and putting the product filtrate into a turnover barrel.
S6: spreading anhydrous sodium sulfate into a product filtrate bucket, fully stirring, and standing for 5-18 h; then placing the mixture into a reaction kettle 3 for normal pressure distillation and concentration, and controlling the kettle temperature to be 70-150 ℃; when the product solution in the kettle is concentrated to be sticky, the product solution is discharged; the solvent is recycled and reused.
S7: drying the concentrated solution in a drying chamber in a split-disc manner, controlling the temperature to be 60-135 ℃, drying for 30-60 h, and removing the solvent to obtain a crude product; then, mixing the crude product and trichloromethane according to the proportion of 1: (3-10) re-dissolving, standing for 5-15 h, performing suction filtration, distilling, concentrating, recovering the solvent, and drying the concentrated solution again to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Furthermore, the applicant consults relevant standard data at home and abroad, and does not consult the standard for testing 2, 6-di-O-methyl-beta-cyclodextrin. Further, the catalogues of several large reagent companies such as Sigam Chemical Company, FluKa Chemika-BichemiKa Company, Aldrich Chemical Company. inc in China were referred to, and these larger foreign manufacturers did not give the same specification index to the product. Besides the molecular formula and molecular weight of the compound, some companies give nuclear magnetic resonance spectrum, carbon element analysis table, Raman spectrum or mass spectrum data and the like of the product, and the indexes for controlling the quality of the product are few. Thus, the present invention further provides a method for assaying 2,6-di-O-methyl- β -cyclodextrin. Specifically, the method comprises the following steps:
subject matter and application scope
The invention specifies [2, 6-di-O-methyl ]]7-beta-cyclodextrin (sometimes abbreviated as 2.6-dimethyl-beta-cyclodextrin), detection rules, inspection methods, packaging, transportation, storage, and the like. The inspection standard for producing the product and applying the product.
(II) Standard name
[ 2.6-di-O-methyl group]7-beta-cyclodextrin, Heptakis (2.6-di-O-methyl) -beta-cyclodextrin is the standard name for this product, sometimes abbreviated as 2.6-di-O-methyl-beta-cyclodextrin or 2.6-dimethyl-beta-cyclodextrin, some foreign documents are often abbreviated as 2.6-di-O-methyl-beta-cyclodextrin.
Reference standard (III)
GB/T619-1988 chemical reagent sampling and acceptance rules
Universal method for measuring GB9741-88 chemical reagent burning residues
GB/T6682-1992 analytical laboratory Water Specifications and test methods
GB15346-1994 Package and Mark
(IV) technical Condition
[ 2.6-di-O-methyl group]7-beta-cyclodextrin of formula: c56H98O35，
Molecular weight: 1331.36
Appearance: white powder
The technical conditions of the product of the invention are as follows:
(V) test rules
According to the GB/T619-1988, at least four sampling points are determined in production before each batch of products is packaged, in order to prevent moisture absorption, a sampling container is a polyethylene small bag or a sealable polyethylene or glass small bottle, the sampling container is immediately closed after the sampling is finished, and a label with a written date and a written batch number is attached. For ready inspection.
(VI) test method
1. Determination of percentage content of carbon and hydrogen elements
The instrument comprises the following steps: the precision and accuracy of the organic element analyzer are within +/-0.3%.
The principle is as follows: the organic compound of the sample is burnt and decomposed under the action of a catalyst at high temperature in an organic element analyzer, and both carbon and hydrogen elements in the compound are quantitatively converted into corresponding small molecular compounds, for example, the carbon element is converted into CO2Conversion of hydrogen element to H2O, the small molecular compounds are separated by a chromatographic column in the instrument, and the instrument automatically bases on CO2And H2And calculating the respective percentage contents of the carbon and the hydrogen elements by the peak area of the O.
And (3) analysis operation: the sample to be analyzed is preferably stored in a desiccator to prevent absorption of water. If the sample absorbs moisture, the sample is put into a weighing bottle, put into an oven, dried for 1 hour at the temperature of 100-105 ℃, then transferred into a dryer, and after the sample is cooled, a certain amount of sample is accurately weighed according to the requirements of the instrument for analysis.
And (3) analysis results: the [ 2.6-di-O-methyl group]7The analysis result of the beta-cyclodextrin sample shows that the carbon content is 49.90-51.10%, and the hydrogen content is 7.10-7.80%.
2. Water dissolution test
0.5 g (2.6-di-O-methyl) is weighed7-beta-cyclodextrin sample, dissolved in 3 ml of distilled water, the solution should be transparent and free of mechanical impurities.
3. Burning residue
Weighing (2.6-di-O-methyl)7The accurate amount of the-beta-cyclodextrin sample (5 +/-0.1) g is 1mg, the sample is placed in a crucible with constant weight at the temperature of 650 +/-50 ℃, then the crucible containing the sample is placed on an electric furnace, the sample is completely carbonized by small fire, and then the crucible is placed in a high-temperature furnace (a resistance furnace) and is slowly heated until the weight of the sample is constant at the temperature of 650 +/-50 ℃, and the weight of residues cannot be more than 5.0 mg.
The mass percentage of the burned residue is calculated according to the following formula:
in the formula: x-mass percent of ignition residue: is based on
m2-mass of residue and empty crucible: gram (g)
m1-mass of empty crucible: gram (g)
m-mass of sample: gram (g)
The following description is made with reference to specific embodiments:
example one
This example provides a method for the industrial mass production of 2,6-di-O-methyl- β -cyclodextrin, comprising the steps of:
s1: drying the beta-cyclic dextrin before feeding, controlling the temperature at 130 ℃, the drying time for 2h and the water loss rate to be 20%.
S2: sequentially adding anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide into the reaction kettle 1, fully stirring, and cooling to 10 ℃ for keeping; the mass ratio of anhydrous barium oxide, barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 30: 50: 40: 35, adding the beta-cyclodextrin by 5 times, separating two adjacent times for 5min, and continuously stirring for 3h after all the beta-cyclodextrin are added; and then dropwise adding methyl sulfate, wherein the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: 10, evenly dividing methyl sulfate into 5 parts, dripping each part for 1.5h, and stirring for 60 min; after all the dropwise addition is finished, stirring and reacting for 10 hours.
S3: and (4) turning off the refrigeration system in the S2, continuously stirring, heating to 30 ℃, and keeping the temperature to continue the reaction for 20 hours.
S4: and (3) extracting the product obtained in the step (S3) with trichloromethane, then placing the extracted product into a reaction kettle 2, stirring, controlling the kettle temperature at 30 ℃, slowly adding a saturated sodium bicarbonate solution for neutralization, continuing stirring for 18h after 1h, standing for layering, and discharging a product solution.
S5: sequentially carrying out salt washing on the neutralized product, washing the neutralized product by adopting saturated salt water for 2 times, controlling the temperature of a kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3 h; then washing with pure water for 2 times, controlling the temperature of the kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3.5 h; and then, carrying out suction filtration on the product solution after washing by using a G4 sand core funnel, and putting the product filtrate into a turnover barrel.
S6: spreading anhydrous sodium sulfate into the product filtrate bucket, fully stirring, and standing for 18 h; then placing the mixture into a reaction kettle 3 for normal pressure distillation and concentration, and controlling the kettle temperature at 70 ℃; when the product solution in the kettle is concentrated to be sticky, the product solution is discharged; the solvent is recycled and reused.
S7: drying the concentrated solution in a drying chamber in a split-disc manner, controlling the temperature at 135 ℃, drying for 30h, and removing the solvent to obtain a crude product; then, mixing the crude product and trichloromethane according to the proportion of 1: 10, re-dissolving, standing for 5 hours, carrying out suction filtration, distilling, concentrating and recovering the solvent, and drying the concentrated solution again to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Example two
This example provides a method for the industrial mass production of 2,6-di-O-methyl- β -cyclodextrin, comprising the steps of:
s1: drying the beta-cyclic dextrin before feeding, controlling the temperature at 90 ℃, the drying time for 15h, and the water loss rate for 3%.
S2: sequentially adding anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide into the reaction kettle 1, fully stirring, and cooling to-10 ℃ for keeping; the mass ratio of anhydrous barium oxide, barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 30: 50: 70: 10, adding the beta-cyclodextrin for 2 times, separating two adjacent times for 30min, and continuously stirring for 0.5h after all the beta-cyclodextrin are added; and then dropwise adding methyl sulfate, wherein the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: 25, evenly dividing the methyl sulfate into 2 parts, dripping the methyl sulfate into the parts for 4 hours, and stirring for 15 min; after all the dropwise addition is finished, stirring and reacting are continued for 25 h.
S3: and (4) turning off the refrigeration system in the S2, continuously stirring, heating to 20 ℃, and keeping the temperature to continue reacting for 60 hours.
S4: and (3) extracting the product obtained in the step (S3) by using trichloromethane, then placing the extracted product into a reaction kettle 2, stirring, controlling the kettle temperature at 10 ℃, slowly adding a saturated sodium bicarbonate solution for neutralization, continuing stirring for 5 hours, standing for layering, and discharging a product solution.
S5: sequentially carrying out salt washing on the neutralized product, washing the neutralized product by adopting saturated salt water for 5 times, controlling the temperature of a kettle to be 10 ℃, stirring for 3 hours each time, and standing and layering for 0.5 hour; then washing with pure water for 5 times, controlling the temperature of the kettle at 10 ℃, stirring for 3.5h each time, and standing for layering for 0.5 h; and then, carrying out suction filtration on the product solution after washing by using a G4 sand core funnel, and putting the product filtrate into a turnover barrel.
S6: spreading anhydrous sodium sulfate into the product filtrate bucket, fully stirring, and standing for 5 h; then placing the mixture into a reaction kettle 3 for normal pressure distillation and concentration, and controlling the kettle temperature at 150 ℃; when the product solution in the kettle is concentrated to be sticky, the product solution is discharged; the solvent is recycled and reused.
S7: drying the concentrated solution in a drying chamber in a split-disc manner, controlling the temperature at 60 ℃, and removing the solvent to obtain a crude product, wherein the drying time is 60 hours; then, mixing the crude product and trichloromethane according to the proportion of 1: 3, re-dissolving, standing for 15h, carrying out suction filtration, distilling, concentrating and recovering the solvent, and drying the concentrated solution again to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
In addition, in order to further highlight the advantages of the technical solution of the present invention, the following comparative examples were provided. The following comparative examples were each prepared by changing the setting of relevant parameters based on example one.
Comparative example 1
This comparative example provides an industrial mass production process for 2,6-di-O-methyl- β -cyclodextrin comprising the steps of:
s1: drying the beta-cyclic dextrin before feeding, controlling the temperature at 130 ℃, the drying time for 2h and the water loss rate to be 20%.
S2: sequentially adding barium hydroxide and N, N-dimethylformamide into the reaction kettle 1, fully stirring, cooling to 10 ℃ and keeping; the mass ratio of barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 50: 40: 35, adding the beta-cyclodextrin by 5 times, separating two adjacent times for 5min, and continuously stirring for 3h after all the beta-cyclodextrin are added; and then dropwise adding methyl sulfate, wherein the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: 10, evenly dividing methyl sulfate into 5 parts, dripping each part for 1.5h, and stirring for 60 min; after all the dropwise addition is finished, stirring and reacting for 10 hours.
S3: and (4) turning off the refrigeration system in the S2, continuously stirring, heating to 30 ℃, and keeping the temperature to continue the reaction for 20 hours.
S4: and (3) extracting the product obtained in the step (S3) with trichloromethane, then placing the extracted product into a reaction kettle 2, stirring, controlling the kettle temperature at 30 ℃, slowly adding a saturated sodium bicarbonate solution for neutralization, continuing stirring for 18h after 1h, standing for layering, and discharging a product solution.
S5: sequentially carrying out salt washing on the neutralized product, washing the neutralized product by adopting saturated salt water for 2 times, controlling the temperature of a kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3 h; then washing with pure water for 2 times, controlling the temperature of the kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3.5 h; and then, carrying out suction filtration on the product solution after washing by using a G4 sand core funnel, and putting the product filtrate into a turnover barrel.
S6: spreading anhydrous sodium sulfate into the product filtrate bucket, fully stirring, and standing for 18 h; then placing the mixture into a reaction kettle 3 for normal pressure distillation and concentration, and controlling the kettle temperature at 70 ℃; when the product solution in the kettle is concentrated to be sticky, the product solution is discharged; the solvent is recycled and reused.
S7: drying the concentrated solution in a drying chamber in a split-disc manner, controlling the temperature at 135 ℃, drying for 30h, and removing the solvent to obtain a crude product; then, mixing the crude product and trichloromethane according to the proportion of 1: 10, re-dissolving, standing for 5 hours, carrying out suction filtration, distilling, concentrating and recovering the solvent, and drying the concentrated solution again to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Comparative example No. two
This comparative example provides an industrial mass production process for 2,6-di-O-methyl- β -cyclodextrin comprising the steps of:
s1: drying the beta-cyclic dextrin before feeding, controlling the temperature at 130 ℃, the drying time for 2h and the water loss rate to be 20%.
S2: sequentially adding anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide into the reaction kettle 1, fully stirring, and cooling to 15 ℃ for keeping; the mass ratio of anhydrous barium oxide, barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 30: 50: 40: 35, adding the beta-cyclodextrin by 5 times, separating two adjacent times for 5min, and continuously stirring for 3h after all the beta-cyclodextrin are added; and then dropwise adding methyl sulfate, wherein the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: 10, evenly dividing methyl sulfate into 5 parts, dripping each part for 1.5h, and stirring for 60 min; after all the dropwise addition is finished, stirring and reacting for 10 hours.
S3: and (4) turning off the refrigeration system in the S2, continuously stirring, heating to 30 ℃, and keeping the temperature to continue the reaction for 20 hours.
S4: and (3) extracting the product obtained in the step (S3) with trichloromethane, then placing the extracted product into a reaction kettle 2, stirring, controlling the kettle temperature at 30 ℃, slowly adding a saturated sodium bicarbonate solution for neutralization, continuing stirring for 18h after 1h, standing for layering, and discharging a product solution.
S5: sequentially carrying out salt washing on the neutralized product, washing the neutralized product by adopting saturated salt water for 2 times, controlling the temperature of a kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3 h; then washing with pure water for 2 times, controlling the temperature of the kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3.5 h; and then, carrying out suction filtration on the product solution after washing by using a G4 sand core funnel, and putting the product filtrate into a turnover barrel.
S6: spreading anhydrous sodium sulfate into the product filtrate bucket, fully stirring, and standing for 18 h; then placing the mixture into a reaction kettle 3 for normal pressure distillation and concentration, and controlling the kettle temperature at 70 ℃; when the product solution in the kettle is concentrated to be sticky, the product solution is discharged; the solvent is recycled and reused.
S7: drying the concentrated solution in a drying chamber in a split-disc manner, controlling the temperature at 135 ℃, drying for 30h, and removing the solvent to obtain a crude product; then, mixing the crude product and trichloromethane according to the proportion of 1: 10, re-dissolving, standing for 5 hours, carrying out suction filtration, distilling, concentrating and recovering the solvent, and drying the concentrated solution again to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Comparative example No. three
This comparative example provides an industrial mass production process for 2,6-di-O-methyl- β -cyclodextrin comprising the steps of:
s1: drying the beta-cyclic dextrin before feeding, controlling the temperature at 130 ℃, the drying time for 2h and the water loss rate to be 20%.
S2: sequentially adding anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide into the reaction kettle 1, fully stirring, and cooling to 10 ℃ for keeping; the mass ratio of anhydrous barium oxide, barium hydroxide, N-dimethylformamide to beta-cyclodextrin is 30: 50: 40: 35, adding beta-cyclodextrin and methyl sulfate, and continuously stirring to react for 10 hours after all the beta-cyclodextrin and the methyl sulfate are added; the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: 10.
s3: and (4) turning off the refrigeration system in the S2, continuously stirring, heating to 30 ℃, and keeping the temperature to continue the reaction for 20 hours.
S4: and (3) extracting the product obtained in the step (S3) with trichloromethane, then placing the extracted product into a reaction kettle 2, stirring, controlling the kettle temperature at 30 ℃, slowly adding a saturated sodium bicarbonate solution for neutralization, continuing stirring for 18h after 1h, standing for layering, and discharging a product solution.
S5: sequentially carrying out salt washing on the neutralized product, washing the neutralized product by adopting saturated salt water for 2 times, controlling the temperature of a kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3 h; then washing with pure water for 2 times, controlling the temperature of the kettle at 30 ℃, stirring for 0.5h each time, and standing for layering for 3.5 h; and then, carrying out suction filtration on the product solution after washing by using a G4 sand core funnel, and putting the product filtrate into a turnover barrel.
S6: spreading anhydrous sodium sulfate into the product filtrate bucket, fully stirring, and standing for 18 h; then placing the mixture into a reaction kettle 3 for normal pressure distillation and concentration, and controlling the kettle temperature at 70 ℃; when the product solution in the kettle is concentrated to be sticky, the product solution is discharged; the solvent is recycled and reused.
S7: drying the concentrated solution in a drying chamber in a split-disc manner, controlling the temperature at 135 ℃, drying for 30h, and removing the solvent to obtain a crude product; then, mixing the crude product and trichloromethane according to the proportion of 1: 10, re-dissolving, standing for 5 hours, carrying out suction filtration, distilling, concentrating and recovering the solvent, and drying the concentrated solution again to obtain the target product 2, 6-di-O-methyl-beta-cyclodextrin.
Further, the target products prepared in each example and comparative example are characterized; see table 1 for details.
TABLE 1 statistical table of physicochemical properties of target products of examples and comparative examples
| Yield/%) | Carbon content/%) | Content of hydrogen/%) | Water dissolution test | Ignition residue/% | |
| Example one | 85 | 50.80 | 7.60 | Qualified | 0.05 |
| Example two | 88 | 51.02 | 7.58 | Qualified | 0.03 |
| Comparative example 1 | 70 | 47.38 | 7.01 | Qualified | 0.08 |
| Comparative example No. two | 75 | 45.69 | 7.95 | Fail to be qualified | 0.13 |
| Comparative example No. three | 63 | 43.59 | 6.97 | Fail to be qualified | 0.15 |
The industrial preparation method of the 2, 6-di-O-methyl-beta-cyclodextrin provided by the invention is initiated at home, breaks through the limitation that the original production can be carried out only in a small amount in a laboratory, successfully solves the current situation that the 2, 6-di-O-methyl-beta-cyclodextrin cannot be industrially prepared in a large amount at home, further has good promotion effects on mass production of pertussis vaccines without side effects and prevention of children pertussis, and has remarkable social benefits.
It is to be noted that, unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the invention pertains. Unless specifically stated otherwise, the relative steps, numerical expressions, and values of the components and steps set forth in these embodiments do not limit the scope of the present invention. In all examples shown and described herein, unless otherwise specified, any particular value should be construed as merely illustrative, and not restrictive, and thus other examples of example embodiments may have different values.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention, and all of the technical solutions are covered in the protective scope of the present invention.
Claims (1)
1. An industrial preparation method of 2, 6-di-O-methyl-beta-cyclodextrin is characterized by comprising the following steps:
s1: drying the beta-cyclic dextrin before feeding until the water loss rate is 3-20%;
s2: mixing anhydrous barium oxide, barium hydroxide and N, N-dimethylformamide under low temperature, and stirring; then adding the dried beta-cyclodextrin and stirring, then dropwise adding methyl sulfate and stirring, and continuing stirring for a first preset time after all dropwise adding is finished;
s3: closing the low-temperature system in the S2, continuously stirring and heating to 20-30 ℃, keeping the temperature and continuously reacting for 20-60 h;
s4: extracting the product obtained in the step S3 by using trichloromethane, and then adding saturated sodium bicarbonate solution for neutralization;
s5: sequentially carrying out salt washing and water washing on the neutralized product, then carrying out suction filtration and collecting filtrate;
s6: dehydrating the filtrate, and then distilling and concentrating the filtrate until the filtrate is concentrated to be viscous;
s7: drying the concentrated solution to remove the solvent to obtain a crude product; then recrystallizing the crude product to obtain a target product 2, 6-di-O-methyl-beta-cyclodextrin;
the step of S7 further includes: crushing the target product 2, 6-di-O-methyl-beta-cyclodextrin to 40-70 meshes;
in said S1: the drying treatment specifically comprises the following steps: the temperature is 90-130 ℃, and the time is 2-15 h;
in said S2: the low temperature is specifically-10 ℃ to 10 ℃; the mass ratio of the anhydrous barium oxide, the barium hydroxide, the N, N-dimethylformamide and the beta-cyclodextrin is 30: 50: (40-70): (10-35); and the mass ratio of the methyl sulfate to the beta-cyclodextrin is 24: (10-25);
adding the beta-cyclodextrin in several times, wherein the interval between every two adjacent times is 5-30 min, and continuously stirring for 0.5-3 h after the beta-cyclodextrin is completely added; adding the methyl sulfate in several times, dropwise adding the methyl sulfate in 1.5-4 h of each part, and continuously stirring for 15-60 min; the first preset time is 10-25 h; adding the beta-cyclodextrin and the methyl sulfate for 1-5 times;
in said S4: the neutralization is specifically as follows: slowly adding a saturated sodium bicarbonate solution into the extracted product at the temperature of 10-30 ℃, continuously stirring for a second preset time, standing for layering, and taking a liquid phase; wherein the saturated sodium bicarbonate solution is added for 1-5 hours, and the second preset time is 5-18 hours;
in said S5: the salt washing specifically comprises the following steps: washing the neutralized product by saturated saline for several times, wherein in the washing process, the temperature is 10-30 ℃, the product is stirred for 0.5-3 h each time, and the product is kept stand for 0.5-3 h for layering; the water washing specifically comprises the following steps: washing the neutralized product for several times by adopting pure water, wherein in the washing process, the temperature is 10-30 ℃, stirring is carried out for 0.5-3.5 h each time, and standing and layering are carried out for 0.5-3.5 h; carrying out salt washing and water washing for 2-5 times uniformly;
in said S6: the dehydration treatment specifically comprises the following steps: spreading anhydrous sodium sulfate into the filtrate, fully stirring, and standing for 5-18 h; in the distillation and concentration, the temperature is 70-150 ℃;
in said S7: the drying process comprises the following steps: the temperature is 60-135 ℃, and the time is 30-60 h; in the recrystallization process: and (3) mixing the crude product with trichloromethane according to the proportion of 1: (3-10) re-dissolving, standing for 5-15 h, performing suction filtration, distilling, concentrating, recovering the solvent, and drying the concentrated solution again.
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| JP2008095056A (en) * | 2006-10-11 | 2008-04-24 | Tokyo Kasei Kogyo Kk | New cyclodextrin derivative |
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| CN104558253A (en) * | 2014-12-19 | 2015-04-29 | 福建工程学院 | Green synthesis method of 2-O-methyl-6-O-(2-hydroxypropyl)-beta-cyclodextrin |
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| JP2008095056A (en) * | 2006-10-11 | 2008-04-24 | Tokyo Kasei Kogyo Kk | New cyclodextrin derivative |
| CN101508741A (en) * | 2009-03-16 | 2009-08-19 | 濮阳职业技术学院 | Process for synthesis of methylation-beta-cyclodextrin with dimethyl carbonate and beta-cyclodextrin |
| CN104558253A (en) * | 2014-12-19 | 2015-04-29 | 福建工程学院 | Green synthesis method of 2-O-methyl-6-O-(2-hydroxypropyl)-beta-cyclodextrin |
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