CN109836397A - A kind of preparation method of biological buffer-morpholinopropane sulfonic acid (MOPS) - Google Patents

Abstract

The invention belongs to the field of organic synthesis, and in particular relates to a preparation method of a biological buffer-morpholinyl propane sulfonic acid (MOPS), comprising the following steps: morpholine and 1,3-propane sultone (1,3-PS) ) reacted to generate morpholino propane sulfonic acid. The raw material used in the method of the present invention is cheaper than the raw material used in the method for synthesizing morpholinopropane sulfonic acid using 1,3-dibromopropane and morpholine, and is beneficial to large-scale production and application. In the present invention, ethanol and the like are used as the solvent, the solvent is easy to remove, and the user's requirement for solvent residue is satisfied. The decolorization in the method of the invention adopts the method of adding a reducing agent and nitrogen protection, which ensures the chromaticity of the MOPS and makes the obtained product with better properties. The preparation process of the invention is simple, the operation is convenient, and the practicability is high.

Description

A kind of biological buffer-the preparation method of morpholino propane sulfonic acid (MOPS)

technical field

The invention belongs to the field of organic synthesis, and in particular relates to a preparation method of a biological buffer-morpholino propane sulfonic acid (MOPS).

Background technique

MOPS is a very versatile biological buffer, which is widely used in cell culture, biological fermentation and biological preparations, such as hepatitis B diagnostic agent.

There are few reports on the synthesis method of MOPS. Anal.Chen.37(1).156-8 1965 (Eng) first reported the synthesis method of MOPS; WO2001085678 reported the synthesis of MOPS with 1,3-dibromopropane and morpholine. method, but the bromide is too expensive and the preparation cost is higher.

Therefore, it is necessary to improve the preparation method of biological buffer-morpholinyl propane sulfonic acid (MOPS).

SUMMARY OF THE INVENTION

In order to improve the above problems, the present invention provides a preparation method of a biological buffer, morpholino propane sulfonic acid (MOPS), comprising the following steps: morpholine and 1,3-propane sultone (1,3-PS) ) reacts to generate morpholino propane sulfonic acid,

The reaction can be carried out in a polar solvent; the polar solvent can be methanol, ethanol, isopropanol, n-butanol, formamide, trifluoroacetic acid, dimethyl sulfoxide, acetonitrile, N,N-diol. Methylformamide, hexamethylphosphoramide, acetic acid, pyridine, tetramethylethylenediamine, acetone, triethylamine, dioxane, tetrahydrofuran, methyl formate, tributylamine, methyl ethyl ketone, ethyl acetate, tris One or more of octylamine, dimethyl carbonate, diethyl ether, isopropyl ether, n-butyl ether, trichloroethylene, diphenyl ether, dichloromethane, chloroform, dichloroethane, toluene, etc., the solvent preferably ethanol;

According to the present invention, before use, the solvent can be pretreated with a decolorizing agent until the UV absorbance at 260 nm and 280 nm are both less than 0.05, and the decolorizing agent can be activated carbon;

According to the present invention, the molar ratio of the morpholine to 1,3-propane sultone may be (1.0-2.0):1, for example (1.02-1.2):1;

The temperature of the reaction may be 15-50°C, such as 20-30°C;

The reaction time can be 1 to 7 hours, preferably 2 hours;

According to the present invention, after the reaction is completed, the obtained product can also be subjected to cooling crystallization, centrifugal separation, redissolving, decolorization, filtration, cooling crystallization, centrifugation, washing and drying;

Preferably, after the completion of the reaction, the temperature can be cooled and crystallized for several hours, and the crude product can be obtained by centrifugal separation;

Preferably, after obtaining the crude product, add the crude product to an appropriate amount of deionized water, stir and heat up to dissolve, add a reducing agent and a decolorizing agent, decolorize for several hours, and filter at a lower temperature; the reducing agent is preferably hydrosulfite, and the decolorizing agent is preferably activated carbon ;

Preferably, the filtrate is heated to dissolve, cooled to crystallize, centrifuged, washed, and dried to obtain the product. The crystallization process can be carried out under the protection of an inert gas, and the inert gas can be nitrogen or argon. The washing is preferably Use polar solvents for washing.

beneficial effect

1) The raw material used in the method of the present invention is cheaper than the raw material used in the method for synthesizing morpholinopropanesulfonic acid using 1,3-dibromopropane and morpholine, which is beneficial to large-scale production and application.

2) The present invention uses ethanol and the like as a solvent, and the solvent is easy to remove, which satisfies the user's requirements for solvent residues.

3) The decolorization in the method of the present invention adopts the method of adding a reducing agent and nitrogen protection, which ensures the chromaticity of the MOPS and makes the obtained product properties better.

4) The preparation process of the present invention is simple, easy to operate, and has high practicability.

Detailed ways

The production method of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following examples are only for illustrating and explaining the present invention, and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the intended protection scope of the present invention.

Unless otherwise stated, the starting materials and reagents used in the following examples are commercially available or can be prepared by known methods.

Example 1

Add 2500L of pretreated absolute ethanol (water content < 0.02%) into a 5000L glass-lined reactor, add 800kg of molten 1,3-PS under stirring, cool down to 25-30°C with stirring, and control the temperature for 4 hours. 582kg of morpholine was added dropwise, reacted at this temperature for 2 hours, lowered to 0° C., incubated for 10 hours for crystallization, centrifuged to obtain 2160kg of wet product, and the alcohol obtained from the mother liquor could be used for recrystallization.

Add 1600L of alcohol to a 3000L glass-lined jar, add 300kg of deionized water, add 1080kg of the wet product obtained in the previous step under stirring, heat up to 70°C, add deionized water to make the wet product completely dissolve, add 1kg of hydrosulfite, 25kg of medicine Use activated carbon, decolorize for 1 hour, filter by pressure, the filtrate enters the crystallizing tank, add 1600L alcohol to wash the tank, and enter the crystallizing tank through the filter press. When it becomes turbid, keep moisturizing crystals for 1 hour, then cool down to 0～5℃, centrifuge, wash twice with 20L absolute ethanol, spin dry, put into rotary vacuum dryer, control temperature below 80℃, vacuum degree above 0.08MPa , dried until the product is qualified to obtain 548kg of product, yield 80% (calculated by 1,3-PS), sampling detection: content 99.8% (alkali titration), and the UV absorbance of 0.5M aqueous solution at 260nm and 280nm are both 0.02.

Example 2

According to the same method of Example 1, add 2500L of absolute ethanol, add 582kg of morpholine, stir and cool down to 20～25 ℃, drip 1,3-PS 800kg during 4 hours, and react at this temperature for 2 hours after dripping, Cool to 0°C, centrifuge to obtain 2180kg of wet product, recrystallize in two batches according to the method of Example 1, finally obtain 554kg of MOPS, yield 81%, sampling detection: content 99.8% (alkali titration), 0.5M aqueous solution in The UV absorbance at 260 nm and 280 nm is both 0.03.

Comparative Example 1

According to the same method as Example 1, refining without adding hydrosulfite, without nitrogen protection for crystallization, without activated carbon treatment with dehydrated ethanol, the UV absorbance at 260 nm of the product was measured to be 0.5, and the UV absorbance at 280 nm was 0.2.

Comparative Example 2

According to the same method as Example 1, the absolute ethanol contains 0.5% water, and the others are carried out according to Example 1. Finally, 450kg of product is obtained, the yield is 65.7%, and the content is 99.6%. The UV absorbance of the 0.1M aqueous solution at 260nm and 280nm is both 0.05.

Comparative Example 3

According to the same method as in Example 1, the morpholine feeding amount was reduced to 485kg, that is, the molar ratio of 1,3-PS and morpholine was 1:1, and the final product was 530kg, the yield was 77.4%, the content was 99.7%, and the 0.5M aqueous solution was in The UV absorbance at 260 nm and 280 nm is both 0.04.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above-described embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims (10)

1. a kind of biological buffer-morpholinopropane sulfonic acid (MOPS) preparation method, which comprises the following steps: Morpholine and 1,3- propane sultone (1,3-PS) reaction generate morpholinopropane sulfonic acid,

2. preparation method as described in claim 1, which is characterized in that the reaction can carry out in polar solvent；It is described Polar solvent can be methanol, ethyl alcohol, isopropanol, n-butanol, formamide, trifluoroacetic acid, dimethyl sulfoxide, acetonitrile, N, N- bis- Methylformamide, hexamethyl phosphoramide, acetic acid, pyridine, tetramethylethylenediamine, acetone, triethylamine, dioxane, tetrahydrofuran, Methyl formate, tri-n-butylamine, methyl ethyl ketone, ethyl acetate, trioctylamine, dimethyl carbonate, ether, isopropyl ether, n-butyl ether, three chloroethenes One of alkene, diphenyl ether, methylene chloride, chloroform, dichloroethanes, toluene etc. are a variety of, and the solvent is preferably ethyl alcohol.

3. preparation method as claimed in claim 1 or 2, which is characterized in that the solvent before use, can with decolorising agent into Row pretreatment to the ultraviolet absorptivity at 260nm and 280nm is respectively less than 0.05, and the decolorising agent can be active carbon.

4. preparation method as claimed in any one of claims 1-3, which is characterized in that the morpholine and 1, in 3- propane sulfonic acid The molar ratio of ester can be (1.0~2.0): 1, such as (1.02~1.2): 1.

5. such as preparation method of any of claims 1-4, which is characterized in that the temperature of the reaction can for 15~ 50 DEG C, such as 20~30 DEG C.

6. preparation method according to any one of claims 1 to 5, which is characterized in that the time of the reaction can be 1~7 Hour, preferably 2 hours.

7. such as preparation method of any of claims 1-6, which is characterized in that after the reaction was completed, can also will obtain Product carry out decrease temperature crystalline, centrifuge separation, weight molten, decoloration, filtering, decrease temperature crystalline, centrifugation, washing, drying.

8. such as preparation method of any of claims 1-7, which is characterized in that after reaction, can be with decrease temperature crystalline A few hours can obtain crude product by centrifugation point.

9. such as preparation method of any of claims 1-8, which is characterized in that after obtaining crude product, crude product is added to suitable Stirring is warming up to dissolution in amount deionized water, adds reducing agent and decolorising agent, decolourizes a few hours, cooling filtering, and the reducing agent is excellent It is selected as sodium hydrosulfite, the decolorising agent is preferably active carbon.

10. preparation method as claimed in any one of claims 1-9 wherein, which is characterized in that by filtrate rising temperature for dissolving, cooling knot Crystalline substance is centrifugated, and is washed, dry, product can be obtained, crystallizing tank can be protected with inert gas, and the inert gas can Think nitrogen or argon gas, it is preferable to use polar solvents to be washed for the washing.