Preparation method of organic ammonium pyrophosphate solid

Abstract

The invention provides a preparation method of a pyrophosphoric acid organic ammonium salt solid, which comprises the following steps: removing water in a pyrophosphoric acid organic ammonium salt solution by an ethanol distillation method under the action of negative pressure, freezing the obtained product, and then freeze-drying to obtain the pyrophosphoric acid organic ammonium salt solid. The method provided by the invention solves the problems that the organic ammonium pyrophosphate is difficult to remove water and high in cost price; the solid of the organic ammonium pyrophosphate can be prepared, storage and transportation are convenient, raw materials are easy to obtain and cheap, and the production cost is greatly reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of organic ammonium pyrophosphate salts, and in particular relates to a method for preparing solid organic ammonium pyrophosphate salts. Background Art

[0002] Natural nucleoside polyphosphates not only play an important role in life activities, but also are widely used as important tools in many life science and medical fields together with their analogs. Among the many nucleoside polyphosphate biomolecules, nucleoside triphosphate (NTP) is an important substance that maintains the operation of the life system. Natural nucleoside triphosphates include: (deoxy) adenosine triphosphate ((d) ATP), (deoxy) guanosine triphosphate ((dGTP), (deoxy) cytidine triphosphate ((d) CTP), uridine triphosphate (UTP), deoxythymidine triphosphate (dTTP). Deoxynucleoside triphosphates (dNTPs) are the basic units that constitute the structure of DNA. With the beginning of the first generation of DNA sequencing technology in 1977, it has developed rapidly in the past forty years. At present, most of the nucleoside triphosphates used in DNA sequencing are chemically synthesized. The more common methods for synthesizing dNTPs are the "one-pot three-step method", "salicylic acid phosphite chloride method" and "phosphoamide method". In the "one-pot three-step method" and the "salicylic acid phosphite chloride method", the nucleoside monophosphorus compound is first formed during the reaction, and then reacted with pyrophosphate to generate nucleoside triphosphate. The pyrophosphate used here mainly refers to tributylammonium pyrophosphate ((Bu3NH)2PPi) or tetra(tributyl)ammonium pyrophosphate ((Bu4N)2PPi).

[0003] The prior art for preparing pyrophosphoric acid mostly uses sodium pyrophosphate or sodium pyrophosphate decahydrate as raw materials, converts sodium pyrophosphate into pyrophosphoric acid through ion exchange resin, and then reacts with an alcohol solution of an organic amine to obtain an aqueous solution of pyrophosphoric acid of the organic amine, and then concentrates most of the water to form a paste-like product. Since the organic ammonium pyrophosphate salt is very easy to absorb water, it is difficult to obtain a dry solid powder. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a method for preparing a pyrophosphate organic ammonium salt solid. The method provided by the present invention can prepare a pyrophosphate organic ammonium salt solid, and the preparation method has a simple process and low cost.

[0005] The present invention provides a method for preparing a solid organic ammonium pyrophosphate salt, comprising:

[0006] Under the action of negative pressure, water in the organic ammonium pyrophosphate salt solution is removed by ethanol distillation, and the obtained product is frozen and then freeze-dried to obtain an organic ammonium pyrophosphate salt solid.

[0007] In an embodiment of the present invention, the method for preparing the organic ammonium pyrophosphate solution may include:

[0008] The pyrophosphoric acid and the alcohol solution of the organic amine are reacted to obtain the pyrophosphate organic ammonium salt solution.

[0009] In an embodiment of the present invention, the method for preparing pyrophosphoric acid may include:

[0010] The pyrophosphate inorganic salt is subjected to ion exchange to obtain pyrophosphate.

[0011] In an embodiment of the present invention, the inorganic pyrophosphate salt may be an inorganic pyrophosphate salt solution, and the inorganic pyrophosphate salt solution may be obtained by dissolving the inorganic pyrophosphate salt in water; the water may be pure water; and the dissolution may be stirring dissolution. In an embodiment of the present invention, the inorganic pyrophosphate salt may be selected from one or more of sodium pyrophosphate and its crystalline hydrate, potassium pyrophosphate and its crystalline hydrate; the ratio of the volume of water to the mass of the inorganic pyrophosphate salt in the inorganic pyrophosphate salt solution may be (10-100) mL:1 g, or (10-50) mL:1 g, or (10-20) mL:1 g.

[0012] In an embodiment of the present invention, the ion exchange may be a cation exchange, and the ion exchange may use a cation exchange resin, which may be a strong acid cation exchange resin, such as one or more of the commercially available product models DOWEX 50, domestic 732, Amberlite IR, and Diaion SK.

[0013] In the embodiment of the present invention, the ion exchange may be to pass the pyrophosphate inorganic salt solution through a strong acid cation resin, and collect the solution with pH < 6 after the solution flows out, so as to obtain pyrophosphate.

[0014] In an embodiment of the present invention, an alcohol solution of an organic amine can be prepared by mixing an alcohol and an organic amine; the mixing can be carried out under stirring; the stirring can be carried out using a magnetic stirrer; the alcohol can be selected from one or more of methanol, ethanol, propanol, n-butanol, and tert-butanol; the organic amine can be selected from one or more of a primary amine, a secondary amine, or a tertiary amine, such as a tertiary amine; the primary amine can be selected from one or more of ethylamine, propylamine, n-butylamine, and tert-butylamine; the secondary amine can be selected from one or more of diethylamine, dipropylamine, di-n-butylamine, and di-tert-butylamine; the tertiary amine can be selected from one or more of triethylamine, tripropylamine, tri-n-butylamine, tri-tert-butylamine, and N,N-diisopropylethylamine, such as tri-n-butylamine. In an embodiment of the present invention, the ratio of the volume of alcohol in the alcohol solution of organic amine to the mass of inorganic pyrophosphate can be (1-10) mL:1 g, or (1-5) mL:1 g; the molar ratio of organic amine to inorganic pyrophosphate in the alcohol solution of organic amine can be (1-5):1, or (2-3):1.

[0015] In an embodiment of the present invention, during the reaction of pyrophosphoric acid and an alcohol solution of an organic amine, pyrophosphoric acid can be dropped into the alcohol solution of an organic amine under stirring; the reaction can be carried out under stirring; the reaction temperature can be -20 to 50°C, -10 to 20°C, or -5 to 10°C; the reaction time can be 1 to 120 min, 10 to 80 min, 10 to 50 min, or 10 to 30 min.

[0016] In an embodiment of the present invention, after the organic ammonium pyrophosphate salt solution is obtained, it can be concentrated to obtain a pasty product, and then the pasty product is distilled using ethanol to remove water therein; the concentration method can be to remove water and alcohol in the organic ammonium pyrophosphate salt solution by reduced pressure distillation under negative pressure conditions; the organic ammonium pyrophosphate salt solution can be connected to a vacuum pump, and the negative pressure condition is achieved by vacuuming the vacuum pump; the vacuum pump can be selected from an oil pump, a water pump or a diaphragm pump, such as an oil pump or a diaphragm pump; the vacuum degree of the negative pressure condition can be 1-20Pa, or 5-15Pa, or 10Pa; the temperature of the reduced pressure distillation can be 0-50°C, or 10-40°C, or 20-30°C; the volume of water and alcohol removed from the organic ammonium pyrophosphate salt solution can be 60-90% of the total volume of the organic ammonium pyrophosphate salt solution, or 80-90%.

[0017] In an embodiment of the present invention, the method of ethanol distillation may include:

[0018] mixing ethanol and an organic ammonium pyrophosphate salt solution to obtain a mixture;

[0019] Under the action of negative pressure, the ethanol in the mixture is distilled to remove the water therein.

[0020] In an embodiment of the present invention, the volume ratio of ethanol to the organic ammonium pyrophosphate solution or the above-mentioned paste product can be (1-10):1, (1-8):1, (1-4):1, or (1-2):1.

[0021] In an embodiment of the present invention, the negative pressure effect can be achieved by a vacuum pump, such as connecting the above-mentioned mixture to a vacuum pump, and placing the above-mentioned mixture in a negative pressure state by the vacuuming effect of the vacuum pump; the vacuum pump can be selected from an oil pump, a water pump or a diaphragm pump, such as an oil pump or a diaphragm pump; the vacuum degree of the negative pressure effect can be 1 to 20Pa, 5 to 15Pa, or 10Pa.

[0022] In an embodiment of the present invention, distillation can be performed using a rotary evaporator, and ethanol can be distilled under reduced pressure by a rotary evaporator under the action of vacuum; the temperature of the reduced pressure distillation can be 0-50°C, 10-40°C, or 20-30°C.

[0023] In an embodiment of the present invention, ethanol is used to distill away water in the paste product to obtain a viscous liquid, and the obtained viscous liquid is frozen and then freeze-dried to obtain a pyrophosphate organic ammonium salt solid.

[0024] In the embodiments of the present invention, freezing can be performed by using a refrigerator; a viscous liquid can be frozen into a solid by using a refrigerator; the freezing temperature can be below 0°C, such as -20 to -80°C, or -40 to -60°C; the freezing time can be 1 to 48 hours, or 12 to 24 hours.

[0025] In the embodiments of the present invention, freeze drying can be performed using a freeze dryer; the freeze drying pressure can be 1 to 20 Pa, 2 to 10 Pa, or 2 to 5 Pa; the freeze drying time can be 10 to 96 h, or 24 to 48 h; the freeze drying temperature can be -20 to -80°C, or -40 to -60°C.

[0026] In an embodiment of the present invention, the preparation method of the organic ammonium pyrophosphate solid may include:

[0027] The inorganic pyrophosphate salt is exchanged with a cation exchange resin to obtain pyrophosphate, which is then reacted with an alcohol solution of an organic amine to obtain a solution of an organic ammonium pyrophosphate salt, which is then concentrated to obtain a paste-like product;

[0028] Add ethanol to the paste-like product to obtain a mixture of ethanol and organic ammonium pyrophosphate salt solution; then connect the mixture to a vacuum pump to remove water in the organic ammonium pyrophosphate salt solution by distillation of ethanol under negative pressure. When the ethanol is completely distilled, place the remaining viscous liquid in a refrigerator, and finally freeze-dry to obtain an organic ammonium pyrophosphate salt solid.

[0029] In an embodiment of the present invention, the method for preparing a solid organic ammonium pyrophosphate salt may include the following steps:

[0030] 1) Take pyrophosphate inorganic salt in a beaker, add purified water and stir thoroughly to dissolve;

[0031] 2) adding the dissolved pyrophosphate inorganic salt solution to the strongly acidic cationic resin, allowing the solution to flow out, and collecting the solution with a pH value less than 6;

[0032] 3) Take another reaction bottle, add a magnetic stirrer, add alcohol and organic amine, and stir;

[0033] 4) dripping the collected solution from step 2) into the flask from step 3) under stirring;

[0034] 5) Take out the reaction flask, connect it to a vacuum pump and distill under reduced pressure to remove water and alcohol to obtain a paste-like product;

[0035] 6) adding ethanol to the reaction flask to obtain a mixture of ethanol and organic ammonium pyrophosphate solution;

[0036] 7) connecting the mixture of ethanol and the organic ammonium pyrophosphate solution to a rotary evaporator, evacuating the mixture to distill the ethanol under reduced pressure to obtain a viscous liquid;

[0037] 8) putting the viscous liquid from step 7) into a refrigerator and freezing it into a solid;

[0038] 9) The solid obtained in step 8) is taken out from the refrigerator, placed in a freeze dryer, and freeze-dried to obtain an organic ammonium pyrophosphate solid.

[0039] If the obtained paste of organic ammonium pyrophosphate is repeatedly hydrated with ethanol and then prepared into a DMF solution for use, the DMF solution of organic ammonium pyrophosphate is not easy to store, and DMF easily absorbs water, which is troublesome to use; if tetrabutylammonium hydroxide solution is used to react with pyrophosphoric acid, and pyridine is used to azeotropically remove water, a solid organic pyrophosphate salt can be obtained, but tetrabutylammonium hydroxide is relatively expensive, high in cost, and not conducive to scale-up production. The present invention provides a method for obtaining a solid organic ammonium pyrophosphate salt at a low price, which solves the problem that the organic ammonium pyrophosphate salt is difficult to remove water and has a high cost; the method provided by the present invention can prepare a solid organic ammonium pyrophosphate salt, which is convenient for storage and transportation, and the raw materials are easy to obtain and cheap, which greatly reduces the production cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 The tributylammonium pyrophosphate prepared in Example 1 1 H-NMR spectrum;

[0041] Figure 2 The tributylammonium pyrophosphate prepared in Example 1 31 P-NMR spectrum. DETAILED DESCRIPTION

[0042] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0043] Example 1

[0044] Take 20g (43mmol) of sodium pyrophosphate decahydrate in a 500mL beaker, add 300mL of pure water and stir thoroughly to dissolve; add the dissolved sodium pyrophosphate solution to the glass column of DOWEX-50WX8 resin, let the solution flow out, and collect the solution with pH < 6. Take a 1000mL flask, add a magnetic stir bar, 70mL of ethanol and 23g (30mL) of tri-n-butylamine, and stir at 0℃.

[0045] The collected liquid with pH < 6 was dripped into the flask containing ethanol and tri-n-butylamine under stirring, and stirred at 0°C for 20 minutes. The reaction flask was taken out, and ethanol and water were removed by vacuum distillation until the volume of the solution was about 30 mL, and a paste product was obtained. 60 mL of ethanol was added to the reaction flask, and the paste product was mixed with ethanol.

[0046] The mixture was connected to a rotary evaporator and vacuumed at room temperature (vacuum degree of 1-20 Pa) to distill ethanol under reduced pressure to obtain a viscous liquid. The viscous liquid was placed in a -80°C refrigerator for 24 hours to freeze and freeze. Then it was placed in a freeze dryer, the vacuum degree was set to 2 Pa, and the freeze drying time was 48 hours, and finally 18 g of a white solid was obtained, with a yield of 76.8%.

[0047] The product prepared in Example 1 of the present invention was subjected to nuclear magnetic resonance detection to obtain tributylammonium pyrophosphate. 1 H-NMR spectra and 31 P-NMR diagram, such as Figure 1 and Figure 2 As shown, it can be seen that the target product, tributylammonium pyrophosphate, was prepared in Example 1.

[0048] The method provided by the present invention utilizes ethanol vacuum distillation to remove most of the water in the organic ammonium pyrophosphate salt solution, and then freezes the product in a low-temperature refrigerator, and then freezes it in a freeze dryer to obtain a solid. The method provided by the present invention can prepare a solid organic ammonium pyrophosphate salt, which is convenient for storage and transportation, and the raw materials are easily available and cheap, greatly reducing the production cost.

[0049] Although the present invention has been described and illustrated with reference to specific embodiments of the present invention, these descriptions and illustrations do not limit the present invention. It will be clearly understood by those skilled in the art that various changes may be made without departing from the true spirit and scope of the present invention as defined by the appended claims, so that specific circumstances, materials, material compositions, materials, methods or processes are suitable for the goals, spirit and scope of the present application. All such modifications are intended to be within the scope of the appended claims. Although the methods disclosed herein have been described with reference to specific operations performed in a specific order, it should be understood that these operations may be combined, subdivided or reordered to form equivalent methods without departing from the teachings of the present invention. Therefore, unless specifically indicated herein, the order and grouping of operations are not limitations of the present application.

Claims

1. A method for preparing a solid organic ammonium pyrophosphate salt, comprising: Under the action of negative pressure, water in the organic ammonium pyrophosphate salt solution is removed by ethanol distillation, and the obtained product is frozen and then freeze-dried to obtain an organic ammonium pyrophosphate salt solid.

2. The preparation method according to claim 1, characterized in that: The vacuum degree of the negative pressure effect is 1-20Pa.

3. The preparation method according to claim 1, characterized in that: The negative pressure effect is achieved by vacuuming with a vacuum pump, and the vacuum pump is selected from an oil pump, a water pump or a diaphragm pump.

4. The preparation method according to claim 1, characterized in that: The ethanol distillation is carried out by vacuum distillation on a rotary evaporator; The temperature of the reduced pressure distillation is 0-50°C.

5. The preparation method according to claim 1, characterized in that: The volume ratio of the ethanol to the organic ammonium pyrophosphate salt solution is (1-10):

1.

6. The preparation method according to claim 5, characterized in that: The volume ratio of the ethanol to the organic ammonium pyrophosphate salt solution is (1-2):

1.

7. The preparation method according to claim 1, characterized in that: The freezing temperature is -20 to -80°C.

8. The preparation method according to claim 1, characterized in that: The freeze drying process is performed at a pressure of 1 to 20 Pa, a time of 10 to 96 hours, and a temperature of -20 to -80°C.

9. The preparation method according to claim 1, characterized in that: The method of removing water from the organic ammonium pyrophosphate solution by ethanol distillation also includes: Under negative pressure, water and alcohol in the organic ammonium pyrophosphate solution are removed by vacuum distillation; The vacuum degree of the negative pressure is 1 to 20 Pa; The temperature of the reduced pressure distillation is 0-50°C.

10. The preparation method according to claim 9, characterized in that: The volume of water and alcohol removed from the organic ammonium pyrophosphate salt solution is 60-90% of the total volume of the organic ammonium pyrophosphate salt solution.

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