CN114570295A - Method for rapidly preparing triethylamine carbonate buffer solution - Google Patents
Method for rapidly preparing triethylamine carbonate buffer solution Download PDFInfo
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- CN114570295A CN114570295A CN202210307390.1A CN202210307390A CN114570295A CN 114570295 A CN114570295 A CN 114570295A CN 202210307390 A CN202210307390 A CN 202210307390A CN 114570295 A CN114570295 A CN 114570295A
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- AFQIYTIJXGTIEY-UHFFFAOYSA-N hydrogen carbonate;triethylazanium Chemical compound OC(O)=O.CCN(CC)CC AFQIYTIJXGTIEY-UHFFFAOYSA-N 0.000 title claims abstract description 92
- 239000007853 buffer solution Substances 0.000 title claims abstract description 46
- 238000000034 method Methods 0.000 title claims abstract description 29
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims abstract description 147
- 238000006243 chemical reaction Methods 0.000 claims abstract description 145
- 235000011089 carbon dioxide Nutrition 0.000 claims abstract description 61
- 239000001569 carbon dioxide Substances 0.000 claims abstract description 43
- 229910002092 carbon dioxide Inorganic materials 0.000 claims abstract description 43
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 105
- 239000000872 buffer Substances 0.000 claims description 29
- 239000007788 liquid Substances 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- 238000005273 aeration Methods 0.000 claims description 22
- 238000009434 installation Methods 0.000 claims description 21
- 239000004575 stone Substances 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 5
- 229910001069 Ti alloy Inorganic materials 0.000 claims description 3
- 230000006978 adaptation Effects 0.000 claims description 3
- 229910052593 corundum Inorganic materials 0.000 claims description 3
- 239000010431 corundum Substances 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 2
- 230000008878 coupling Effects 0.000 claims description 2
- 238000010168 coupling process Methods 0.000 claims description 2
- 238000005859 coupling reaction Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims 1
- 238000002360 preparation method Methods 0.000 abstract description 20
- 230000035484 reaction time Effects 0.000 abstract description 11
- 230000000694 effects Effects 0.000 abstract description 6
- 238000004904 shortening Methods 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 2
- HWCKGOZZJDHMNC-UHFFFAOYSA-M tetraethylammonium bromide Chemical compound [Br-].CC[N+](CC)(CC)CC HWCKGOZZJDHMNC-UHFFFAOYSA-M 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 24
- 239000007789 gas Substances 0.000 description 16
- 238000005070 sampling Methods 0.000 description 9
- 238000010586 diagram Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 239000002912 waste gas Substances 0.000 description 6
- 239000012295 chemical reaction liquid Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 230000000630 rising effect Effects 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 230000005587 bubbling Effects 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
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- 125000003729 nucleotide group Chemical group 0.000 description 2
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- 238000000926 separation method Methods 0.000 description 2
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- 231100000331 toxic Toxicity 0.000 description 2
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- 238000009423 ventilation Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000006177 biological buffer Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
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- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000012495 reaction gas Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- BFKJFAAPBSQJPD-UHFFFAOYSA-N tetrafluoroethene Chemical group FC(F)=C(F)F BFKJFAAPBSQJPD-UHFFFAOYSA-N 0.000 description 1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J10/00—Chemical processes in general for reacting liquid with gaseous media other than in the presence of solid particles, or apparatus specially adapted therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/0053—Details of the reactor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/18—Stationary reactors having moving elements inside
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00049—Controlling or regulating processes
- B01J2219/00051—Controlling the temperature
- B01J2219/00121—Controlling the temperature by direct heating or cooling
- B01J2219/00123—Controlling the temperature by direct heating or cooling adding a temperature modifying medium to the reactants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00049—Controlling or regulating processes
- B01J2219/00177—Controlling or regulating processes controlling the pH
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/141—Feedstock
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Abstract
The embodiment of the invention provides a method for rapidly preparing triethylamine carbonate buffer solution, and relates to the technical field of buffer solution preparation. The method for rapidly preparing the triethylamine carbonate buffer solution comprises the step of adding dry ice for reducing the temperature of a reaction system and participating in reaction into the reaction system for preparing the triethylamine carbonate buffer solution. It can play the effect of effectively reducing reaction system temperature through adding the dry ice to reaction system, has still played the effect of supplementary carbon dioxide in order to accelerate the formation of triethylamine carbonate simultaneously, very big shortening reaction time, and then make this application can accomplish the preparation to TEAB buffer solution under the ordinary pressure state fast.
Description
Technical Field
The invention relates to the technical field of buffer solution preparation, in particular to a method for quickly preparing triethylamine carbonate buffer solution.
Background
Triethylamine carbonate (TEAB) is widely used for preparing protein biological buffer and mobile phase of liquid chromatography, especially for separating and identifying oligonucleotide. With the rapid development of nucleic acid detection and nucleic acid drug technology in recent years, the demand for analysis and purification of nucleoside, nucleotide and oligonucleotide is rising day by day, and TEAB is used as a gaseous salt buffer solution, which can be removed by reduced pressure distillation without additional desalting operation. This property makes TEAB widely used in the purification preparation process of nucleic acids, but commercial 1M TEAB (pH 8) buffer is up to 1000 yuan per 100 mL. As a research and development department of the production process of rare modified nucleotide for mRNA, hundreds of liters of 1M TEAB stock solution is consumed every day, and the high purchase cost of the purified stock solution is difficult to bear. However, the main raw material of high-purity triethylamine for preparing the TEAB stock solution only needs 100 yuan/L, so that the TEAB stock solution is always prepared independently for departments needing a large amount of TEAB buffer solution.
In general, in the laboratory configuration of TEAB, carbon dioxide gas is introduced into a triethylamine aqueous solution beaker for bubbling for 5-6 hours in an ice bath environment until the pH value is reduced to 8, and then the method is used, but the method is extremely long in time consumption, and a large amount of toxic and pungent triethylamine gas can overflow to harm the health of liquid configuration personnel.
In view of this, the present application is specifically made.
Disclosure of Invention
Objects of the present invention include, for example, providing a method for rapidly preparing a triethylamine carbonate buffer, which can simplify and accelerate the preparation of the triethylamine carbonate buffer.
Embodiments of the invention may be implemented as follows:
in a first aspect, the present invention provides a method for rapidly preparing a triethylamine carbonate buffer, which comprises adding dry ice having a function of reducing the temperature of a reaction system and participating in a reaction to the reaction system for preparing the triethylamine carbonate buffer.
In an alternative embodiment, water, triethylamine and carbon dioxide are included in the reaction system, and the dry ice is added before the water, the triethylamine and the carbon dioxide are mixed;
preferably, the water and the triethylamine are added firstly, then the dry ice is added into the mixed solution of the water and the triethylamine, and then the carbon dioxide is introduced for reaction;
preferably, the dry ice is added first, then the water and the triethylamine are added, and then the carbon dioxide is introduced for reaction. In an alternative embodiment, adding the dry ice prior to the reacting comprises: water and triethylamine are added into the reactor, then the dry ice is added, and then carbon dioxide is introduced for reaction.
In an alternative embodiment, the dry ice is added in an amount of 10-30% by mass of the total mass of the liquid of the water and the triethylamine.
In alternative embodiments, the dry ice is in the form of pellets, blocks, or cylinders;
preferably, the dry ice has a weight of 10-15 g/piece.
In an alternative embodiment, the carbon dioxide has an inlet pressure of 0.1 to 0.2 mpa;
preferably, the inlet flow rate of the carbon dioxide is 1-3L/min.
In an alternative embodiment, the pH value of the reaction system is monitored in real time by a pH detector inserted in the reaction system, and the reaction is terminated when the pH value of the reaction system is 7 to 8.5.
In an alternative embodiment, the reaction system is placed in a reactor for preparing triethylamine carbonate buffer solution for reaction; introducing the carbon dioxide by using an air inlet pipe communicated with the reactor, and detecting the pH of a reaction system in the reactor by using a pH detector arranged in the reactor;
preferably, the tail end of the air inlet pipe is provided with an aeration piece;
preferably, the aeration member is corundum aeration stone or titanium alloy aeration stone.
In an alternative embodiment, exhaust gas generated in the reaction process is discharged by using an exhaust pipe communicated with the reactor;
preferably, the blast pipe includes that exhaust installation pipe and exhaust are outer to be managed, exhaust installation pipe with the opening adaptation of reactor, the pipe diameter of exhaust installation pipe is greater than the pipe diameter of intake pipe, the intake pipe passes the lateral wall of exhaust installation pipe and inserts in the reactor, the outer union coupling of exhaust is to on the lateral wall of exhaust installation pipe.
In an alternative embodiment, the reactor is arranged on a magnetic stirrer, and a magnetic stirrer arranged in the reactor is used for stirring a reaction system;
preferably, the rotation speed of the magnetic stirrer is 500-1200 rpm.
In an alternative embodiment, the reaction system after the reaction is discharged by using a drain pipe arranged at the bottom of the reactor, and the reaction system is filtered by using a filter connected to the drain pipe;
preferably, the filter is a 0.22 μm capsule filter.
The beneficial effects of the embodiment of the invention include, for example:
the embodiment of the invention provides a method for quickly preparing a triethylamine carbonate buffer solution, which has the advantages that dry ice is added into a reaction system, so that the effect of effectively reducing the temperature of the reaction system can be achieved, the effect of supplementing carbon dioxide to accelerate the generation of triethylamine carbonate can be achieved, the reaction time is greatly shortened, and the preparation of the TEAB buffer solution can be quickly completed under the normal pressure state.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a schematic view of an installation method of an air inlet pipe, an air outlet pipe and a pH detector of a reactor for preparing triethylamine carbonate buffer solution provided by the present application;
FIG. 2 is a top view of FIG. 1;
FIG. 3 is a schematic view of another installation manner of an air inlet pipe, an air outlet pipe and a pH detector of the reactor for preparing triethylamine carbonate buffer solution provided by the present application;
FIG. 4 is a top view of FIG. 3;
FIG. 5 is a schematic diagram of a reactor for preparing triethylamine carbonate buffer solution provided by the present application, wherein the reactor is provided with a dry ice feeding pipe;
FIG. 6 is a diagram showing the reaction process for preparing triethylamine carbonate buffer solution according to example 1 of the present application;
FIG. 7 is a diagram showing the reaction process for preparing triethylamine carbonate buffer solution according to example 2 of the present application;
FIG. 8 is a diagram showing the reaction progress for preparing triethylamine carbonate buffer solution according to comparative example 1 of the present application;
FIG. 9 is a diagram showing the reaction progress for preparing triethylamine carbonate buffer solution according to comparative example 2 of the present application;
FIG. 10 is a diagram showing the reaction progress for preparing triethylamine carbonate buffer solution according to comparative example 3;
FIG. 11 is a diagram showing the reaction progress for preparing triethylamine carbonate buffer solution according to comparative example 4 of the present application.
Icon: 100-a reactor for preparing triethylamine carbonate buffer solution; 110-a reaction flask; 111-bottle cap; 1111-mounting holes; 112-an air inlet pipe; 1121-aeration member; 113-exhaust pipe; 1131 — exhaust mounting tube; 1132 — an exhaust extension pipe; 114-a pH detector; 115-drain pipe; 1151-liquid outlet tap; 116-dry ice feed tube; 117-dry ice; 118-magnetic stirrer.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In the description of the present invention, it should be noted that if the terms "upper", "lower", "inside", "outside", etc. indicate an orientation or a positional relationship based on that shown in the drawings or that the product of the present invention is used as it is, this is only for convenience of description and simplification of the description, and it does not indicate or imply that the device or the element referred to must have a specific orientation, be constructed in a specific orientation, and be operated, and thus should not be construed as limiting the present invention.
Furthermore, the appearances of the terms "first," "second," and the like, if any, are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance.
It should be noted that the features of the embodiments of the present invention may be combined with each other without conflict.
Referring to fig. 1, this example provides a method for rapidly preparing a triethylamine carbonate buffer, which includes adding dry ice 117 for reducing the temperature of the reaction system and participating in the reaction system for preparing the triethylamine carbonate buffer.
The triethylamine is toxic corrosive class A liquid, the flash point of the triethylamine is-7 ℃, so the whole preparation process needs to be carried out under the low temperature condition, the conventional method is directly carried out by placing a reaction system for preparing the triethylamine carbonate buffer solution in an ice bath, however, in the application, dry ice 117 is directly added into the reaction system for preparing the triethylamine carbonate buffer solution, and the reaction is realized under the low temperature condition by utilizing the dry ice 117, so the temperature of the whole reaction process is maintained under the low temperature state. In the application, the temperature of the reaction system can be reduced by adding the dry ice 117, and the generation of triethylamine carbonate can be further accelerated by carbon dioxide generated by sublimating the dry ice 117, so that the configuration work of the triethylamine carbonate buffer solution is simplified and accelerated.
In the present application, there are many possibilities of adding the water, triethylamine and carbon dioxide to the reaction system at the same time, for example, adding them together before the reaction, adding a part before the reaction, adding the rest during the reaction, or adding them gradually directly during the reaction. Preferably, dry ice should be added before water, triethylamine and carbon dioxide are mixed; therefore, the reaction system can be maintained in a lower required temperature range in the initial stage of the reaction, and the whole reaction can be conveniently kept to be carried out in a low-temperature environment in the follow-up process.
Specifically, water and triethylamine may be added first, then dry ice is added to the mixed solution of water and triethylamine, and then carbon dioxide is introduced for reaction; it is also possible to carry out the reaction by first adding dry ice, then adding water and triethylamine, and subsequently introducing carbon dioxide. The volume ratio of triethylamine to water may be selected according to the concentration of TEAB to be formulated, for example in the case of 2M TEAB the volume ratio of triethylamine to water is 1: 2.57.
specifically, in the present application, water and triethylamine are added, then dry ice 117 is added, and then carbon dioxide is introduced to carry out the reaction. The amount of dry ice 117 added is 10-30% of the total mass of the liquid of water and triethylamine. The dry ice 117 is granular, blocky or columnar; preferably, the dry ice 117 weighs 10-15 g/piece. The inventor researches and discovers that when the dry ice 117 is too small, the liquid is sprayed out due to violent reaction caused by adding the dry ice into the reaction system, and the liquid at the bottom of the reactor is quickly frozen and is very difficult to control. When the dry ice 117 is too large, the dry ice 117 is not easily put into the reaction system and is inconvenient to use. Therefore, in the present application, by limiting the weight of the dry ice 117 to the above range, it is possible to avoid that the dry ice 117 is too small to be sublimated completely quickly in a short time, and thus the temperature in the later stage of the reaction cannot be maintained under a low temperature condition.
In the whole reaction process, in order to ensure the uniform mixing of the reaction system, the reaction system is stirred at the stirring speed of 500-1200 rpm. Meanwhile, controlling the pressure of the inlet gas to be 0.1-0.2mpa when the carbon dioxide is introduced; the air inlet flow is 1-3L/min. The inlet pressure and the inlet flow can ensure the inlet amount of carbon dioxide, thereby being beneficial to promoting the reaction process. In addition, in the reaction process of the application, the pH value of the reaction system is monitored in real time by using a pH detector inserted in the reaction system, and the reaction is terminated when the pH value of the reaction system is 7-8.5. The setting of the pH detector can monitor the preparation process of the buffer solution in real time, and the reaction can be immediately stopped when the pH value reaches the corresponding reading without sampling for many times through a sampling port. After the preparation is finished, the reaction system is discharged, and under some special conditions, the discharged reaction system can be filtered by a filter, the filter can effectively filter a small amount of residues or impurities such as microorganisms mixed in the reaction system, and the filtered TEAB buffer solution can be directly used for separation and purification of various biological experiments and chromatographic systems.
In order to more clearly illustrate the present application, an apparatus for implementing the above method for rapidly preparing a triethylamine carbonate buffer is also provided.
In this embodiment, which shows a typical but non-limiting example, the reactor 100 for preparing triethylamine carbonate buffer solution comprises a reaction flask 110, an air inlet pipe 112, an air outlet pipe 113 and a pH detector 114.
In the present application, a mounting hole 1111 is formed in the bottle cap 111 to penetrate through the upper and lower surfaces thereof, and the air inlet pipe 112, the exhaust pipe 113, and the pH detector 114 are inserted into the reaction flask 110 through the mounting hole 1111.
The air inlet pipe 112 is used for introducing carbon dioxide into the reaction flask 110, one end of the air inlet pipe 112 is inserted into the reaction flask 110, and an aeration member 1121 is disposed at the end of the air inlet pipe 112. In the present application, the position of the end of the aeration member 1121 is not limited as long as it is located below the liquid level of the reaction system in the reaction flask 110 so as to provide the reaction system with the reaction gas in the form of aeration. For example, the present application shows a typical but non-limiting example, the aeration members 1121 are located at the bottom of the reaction flask 110. The aeration member 1121 in the present application includes, but is not limited to, corundum aeration stone or titanium alloy aeration stone, and the aeration member 1121 may atomize carbon dioxide into very small bubbles, and the bubbles rapidly react with water and triethylamine while rising to form triethylamine carbonate to prepare a TEAB buffer. The process of reaction can be accelerated through the arrangement of the aeration pieces 1121 in the application, and pressure is not particularly applied, so that the TEAB buffer solution can be rapidly prepared under the normal pressure condition.
The exhaust pipe 113 is used for exhausting gas in the reaction flask 110, and the exhaust pipe 113 is inserted into the reaction flask 110 through the mounting hole 1111. For example, the present application shows a typical but non-limiting example, where the end of the exhaust pipe 113 is located at the top of the reaction flask 110. In this application, the exhaust pipe 113 specifically includes the exhaust installation pipe 1131 and the exhaust outer pipe 1132, the exhaust installation pipe 1131 and the mounting hole 1111 adaptation, the pipe diameter of the exhaust installation pipe 1131 is great, is significantly greater than the pipe diameter of the intake pipe 112, the intake pipe 112 passes through the lateral wall of the exhaust installation pipe 1131 and inserts in the reaction flask 110 through the mounting hole 1111, the pH detector 114 is located in the exhaust installation pipe 1131 (please refer to fig. 1 and fig. 2 in combination), and the exhaust outer pipe 1132 is connected to the lateral wall of the exhaust installation pipe 1131. In this application, through setting up the great exhaust installation pipe 1131 of pipe diameter to set up intake pipe 112 in exhaust installation pipe 1131, can effectively avoid offering the through-hole on bottle lid 111 again when intake pipe 112 inserts the reactor, the trompil number on the bottle lid 111 that reduces, and then reduce waste gas and follow the excessive possibility in other trompil gaps, guaranteed that waste gas only discharges from blast pipe 113.
The pH detector 114 is inserted into the reactor, and the position of the end of the pH detector 114 is not limited as long as the pH detector does not interfere with the aeration member 1121, for example, the present application shows a typical but non-limiting example, the pH detector 114 can be located in the middle of the reaction flask 110, and in the present application, the pH detector 114 is mainly used for detecting the pH of the reaction system in the reactor, so as to perform real-time monitoring of the buffer solution preparation process, and the reaction can be terminated immediately when the pH reaches a corresponding reading value, without sampling through a sampling port for multiple times.
It should be understood that the specific structures and the arrangement positions of the air inlet pipe 112, the air outlet pipe 113, the liquid outlet pipe 115 and the pH detector 114 are not particularly limited in this application, as long as the purposes of air inlet, air outlet, liquid outlet and pH real-time monitoring can be achieved, and the installation thereof can be adaptively adjusted. Referring to fig. 3 and 4, fig. 3 and 4 show that the air inlet pipe 112, the air outlet pipe 113 and the pH detector 114 are directly inserted into the reaction flask 110, and the number of the mounting holes 1111 is three.
The bottom of this application reaction bottle 110 is provided with fluid-discharge tube 115, installs out liquid tap 1151 on fluid-discharge tube 115, goes out liquid tap 1151 and is glass or tetrafluoroethylene tap. The liquid discharge pipe 115 is arranged to discharge the reaction liquid in the reaction bottle 110 after the reaction is finished, so that the liquid is prevented from being poured out by moving the reagent bottle, and the operation is convenient and labor-saving.
Further, the reactor 100 for preparing triethylamine carbonate buffer provided by the present application further includes a filter (not shown), and the filter is in communication with the outlet tap 1151. The filter is a 0.22-micron bag filter, the filter can effectively filter a small amount of residues or microorganisms and other impurities mixed in the reaction liquid, and the filtered TEAB buffer solution can be directly used for separation and purification of various biological experiments and chromatographic systems.
In addition, in order to keep the mixing of the reaction system uniform during the reaction process, the reactor is arranged on the magnetic stirrer, and the reaction system is stirred by the magnetic stirrer 118 arranged in the reactor, and the rotation speed of the magnetic stirrer 118 is 500-1200 rpm.
This application can adopt above-mentioned reactor 100 of preparation triethylamine carbonate buffer solution to realize the quick preparation to triethylamine carbonate buffer solution, on the one hand, this application directly adds dry ice 117 to the reactor 100 of preparation triethylamine carbonate buffer solution, can play the effect of effectively reducing reaction system temperature, the effect of supplementary carbon dioxide in order to accelerate the formation of triethylamine carbonate has still been played simultaneously, so very big shortening reaction time, and then make this application can accomplish the preparation to TEAB buffer solution under the ordinary pressure state fast. Meanwhile, an aeration piece 1121 arranged at the tail end of the air inlet pipe 112 is matched to perform aeration operation on the carbon dioxide, so that the carbon dioxide is atomized into tiny bubbles, and the bubbles rapidly react with water and triethylamine to form triethylamine carbonate to prepare TEAB buffer solution while rising. In addition, the reaction process is detected in real time by the pH detector 114, the reaction can be stopped immediately when the pH value reaches a corresponding reading value, the sampling through a sampling port is not needed for multiple times, the operation is simpler, the whole configuration process is safe and convenient, and the speed is higher than that of the like products.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
Taking 4L 2M triethylamine carbonate (pH 8.5) buffer as an example, this example provides a method for rapidly preparing triethylamine carbonate buffer, which includes the following steps:
(1) a reaction flask 110 was placed above a magnetic stirrer, 1.12L of triethylamine and 2.88L of ultrapure water were added to the reaction flask 110, then, all of block-shaped dry ice 117(10 g/block) was added to the reaction flask 110 in 10 portions, a total of 1200g was obtained, a bottle cap 111 was closed, and the magnetic stirrer was opened to set a rotation speed of 1000rpm to start stirring.
(2) A hose for a carbon dioxide gas path is connected into the gas inlet pipe 112, the gas outlet pipe 113 is directly placed into a ventilation cabinet or a small waste gas filtering carbon tank for waste gas adsorption, the carbon dioxide gas path is opened to adjust the gas inlet pressure to 0.1-0.2mpa, the gas inlet flow of carbon dioxide is adjusted to be 2L/min to bubble liquid in the reactor, and the pH detector 114 is opened to monitor the liquid in the reaction bottle 110 in real time. And after triethylamine and water are completely mutually soluble and the pH value reaches 8.5, closing a carbon dioxide gas supply valve, stopping bubbling, and recording the time, wherein the reaction time is 25 min.
(3) After the reaction is finished, the liquid outlet faucet 1151 at the bottom of the reaction bottle 110 is opened to discharge the prepared TEAB buffer solution, and the liquid outlet faucet 1151 is connected to a 0.22 mu m bag filter through a hose for filtration.
The prepared TEAB buffer was a colorless, transparent, viscous liquid, and the pH was measured at 8.5 and the Conductivity at 25.0-26.0 mS/cm. Consistent with the properties and measurements of commercial TEAB.
Example 2
Taking 4L 2M triethylamine carbonate (pH 8.5) buffer as an example, this example provides a method for rapidly preparing triethylamine carbonate buffer, which includes the following steps:
(1) the reaction flask 110 was placed above a magnetic stirrer, 1.12L of triethylamine and 2.88L of ultrapure water were added to the reaction flask 110, and then all the block-shaped dry ice 117(10-15 g/block) was added to the reaction flask 110 in 10 portions, totaling 800g, and the bottle cap 111 was closed, and stirring was started by opening the magnetic stirrer at 600 rpm.
(2) The carbon dioxide gas path is connected with the air inlet pipe 112 through a hose, the air outlet pipe 113 is directly placed in a ventilation cabinet or connected with a small waste gas filtering carbon tank for waste gas adsorption, the carbon dioxide gas path is opened to adjust the air inlet pressure to 0.2mpa, the air inlet flow of carbon dioxide is adjusted to be 1.5L/min to bubble liquid in the reactor, and the pH detector 114 is opened to monitor the liquid in the reaction bottle 110 in real time. And after triethylamine and water are completely mutually soluble and the pH value reaches 8.5, closing a carbon dioxide gas supply valve, stopping bubbling, and recording the time, wherein the reaction time is 36 min.
(3) After the reaction is finished, the liquid outlet faucet 1151 at the bottom of the reaction bottle 110 is opened to discharge the prepared TEAB buffer solution, and the liquid outlet faucet 1151 is connected to a 0.22 mu m bag filter through a hose for filtration.
The prepared TEAB buffer was a colorless, transparent, viscous liquid, and the pH was measured at 8.5 and the Conductivity at 25.0-26.0 mS/cm. Consistent with the properties and measurements of commercial TEAB.
Comparative example 1
In this comparative example, the reactor was placed directly in an ice-water bath without dry ice 117, and the other parameter conditions were the same as in example 1. The reaction completion time was 160 min.
Comparative example 2
This comparative example is substantially the same as example 1 except that in example 1, the dry ice 117 was added all before the start of the reaction, and in this comparative example, the dry ice 117 was continuously replenished during the reaction by opening the cap, specifically, in this comparative example, the dry ice 117 was added 10 times, and the second time was added after 1min from each addition. The reaction completion time was 27 min.
The prepared TEAB buffer was a colorless, transparent, viscous liquid, and the pH was measured at 8.5 and the Conductivity at 25.0-26.0 mS/cm. Consistent with the properties and measurements of commercial TEAB.
Comparative example 3
This comparative example is substantially the same as example 1 except that the amount of dry ice 117 added in this comparative example is 300 g. The reaction completion time was 46 min.
The prepared TEAB buffer was a colorless, transparent, viscous liquid, and the pH was measured at 8.5 and the Conductivity at 25.0-26.0 mS/cm. Consistent with the properties and measurements of commercial TEAB.
Comparative example 4
This comparative example is substantially the same as example 1 except that the amount of dry ice 117 added in this comparative example is 2000 g. However, since the amount of dry ice charged at one time is too large, the temperature rapidly decreases, resulting in freezing of the liquid at the bottom of the reaction vessel, and the reaction has to be terminated.
Comparative example 5
This comparative example is essentially the same as example 1 except that in this comparative example the dry ice 117 was changed to a rice-like weight of 1-2 g/piece, totaling 1200 g. Because the grain-shaped dry ice has small granularity, the dry ice can be rapidly melted to release a large amount of gas, so that the inside of the reactor is vigorously bubbled, the solution overflows, and the solution preparation fails.
The preparation processes of examples 1-2 and comparative examples 1-5 were monitored, and the progress of the reaction was counted every 5min, and the final preparation process is shown in FIGS. 6-11.
As can be seen from fig. 6-11, the required amount of dry ice 117 is added into the reactor before the reaction, so that the reaction system can effectively reduce the temperature of the reaction system, and carbon dioxide is supplemented to accelerate the generation of triethylamine carbonate, thereby greatly shortening the reaction time. Whereas in comparative example 1, the reaction time was about 160min with a conventional ice-water bath, whereas the reaction time was about 25min for example 1, it can be seen that example 1 significantly reduced the reaction time. Further, as can be seen from comparative example 2, adding 1200g of dry ice ten times does not significantly prolong the buffer solution preparation time, but adding dry ice by continuously opening and closing the upper cover of the reactor is troublesome, which is not favorable for large-scale preparation of the reaction solution. As can be seen from comparative examples 3 and 4, too small an amount of dry ice 117 added easily causes a large extension of the reaction time, or too much easily causes freezing of the bottom of the reactor and a failure in solution arrangement. As can be seen from comparative example 5, when the weight of each piece of dry ice 117 was small, the sublimation time of the dry ice 117 was shortened, and it was likely that the liquid in the reactor was vigorously boiled and the reaction liquid overflowed, and the preparation of the TEAB reaction liquid could not be completed.
In summary, the embodiment of the present invention provides a method for rapidly preparing a triethylamine carbonate buffer solution, in which dry ice 117 is added into a reactor 100 for preparing the triethylamine carbonate buffer solution, so as to effectively reduce the temperature of a reaction system, and supplement carbon dioxide to accelerate the generation of triethylamine carbonate, thereby greatly shortening the reaction time. Meanwhile, an aeration piece 1121 arranged at the tail end of the air inlet pipe 112 is matched to perform aeration operation on the carbon dioxide, so that the carbon dioxide is atomized into tiny bubbles, and the bubbles rapidly react with water and triethylamine to form triethylamine carbonate to prepare TEAB buffer solution while rising. In addition, the reaction process is detected in real time by the pH detector 114, the reaction can be immediately stopped when the pH value reaches a corresponding reading value, multiple sampling through a sampling port is not needed, the operation is simpler, and the conditions of reaction system pollution and the like caused by multiple sampling are effectively avoided.
The above description is only for the specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. A method for rapidly preparing triethylamine carbonate buffer solution is characterized by comprising the step of adding dry ice which is used for reducing the temperature of a reaction system and participates in reaction into the reaction system for preparing the triethylamine carbonate buffer solution.
2. The method for rapidly preparing triethylamine carbonate buffer according to claim 1, wherein the reaction system comprises water, triethylamine and carbon dioxide, and the dry ice is added before the water, the triethylamine and the carbon dioxide are mixed;
preferably, the water and the triethylamine are added firstly, then the dry ice is added into the mixed solution of the water and the triethylamine, and then the carbon dioxide is introduced for reaction;
preferably, the dry ice is added first, then the water and the triethylamine are added, and then the carbon dioxide is introduced for reaction.
3. The method for rapidly preparing triethylamine carbonate buffer according to claim 2, wherein the amount of dry ice added is 10-30% of the total mass of the liquids of water and triethylamine.
4. The method for rapidly preparing triethylamine carbonate buffer according to claim 3, wherein said dry ice is in the form of particles, blocks or columns;
preferably, the dry ice has a weight of 10-15 g/piece.
5. The method for rapidly preparing triethylamine carbonate buffer according to claim 2, wherein the inlet pressure of carbon dioxide is 0.1-0.2 mpa;
preferably, the inlet flow rate of the carbon dioxide is 1-3L/min.
6. The method for rapidly preparing triethylamine carbonate buffer according to claim 2, wherein the pH value of the reaction system is monitored in real time by using a pH detector inserted into the reaction system, and the reaction is terminated when the pH value of the reaction system is 7-8.5.
7. The method for rapidly preparing triethylamine carbonate buffer according to claim 3, wherein the reaction system is placed in a reactor for preparing triethylamine carbonate buffer for reaction; introducing the carbon dioxide by using an air inlet pipe communicated with the reactor, and detecting the pH of a reaction system in the reactor by using a pH detector arranged in the reactor;
preferably, an air exposure piece is arranged at the tail end of the air inlet pipe;
preferably, the aeration member is corundum aeration stone or titanium alloy aeration stone.
8. The method for rapidly preparing triethylamine carbonate buffer according to claim 7, wherein exhaust gas generated during the reaction is discharged through an exhaust pipe connected to the reactor;
preferably, the blast pipe includes that exhaust installation pipe and exhaust are outer to be managed, exhaust installation pipe with the opening adaptation of reactor, the pipe diameter of exhaust installation pipe is greater than the pipe diameter of intake pipe, the intake pipe passes the lateral wall of exhaust installation pipe and inserts in the reactor, the outer union coupling of exhaust is to on the lateral wall of exhaust installation pipe.
9. The method for rapidly preparing triethylamine carbonate buffer according to claim 7, wherein the reactor is arranged on a magnetic stirrer, and a magnetic stirrer arranged in the reactor is used for stirring a reaction system;
preferably, the rotation speed of the magnetic stirrer is 500-1200 rpm.
10. The method for rapidly preparing triethylamine carbonate buffer according to claim 7, wherein the reaction system after the reaction is discharged by using a drain pipe disposed at the bottom of the reactor, and the reaction system is filtered by using a filter connected to the drain pipe;
preferably, the filter is a 0.22 μm capsule filter.
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Application publication date: 20220603 |