阅读说明：本技术 液相色谱法测定钴胺素光化动力学装置及方法 (Device and method for measuring photochemical kinetics of cobalamin by liquid chromatography ) 是由 陈慧 韦莉贞 李霞 于 2021-07-12 设计创作，主要内容包括：本发明公布了一种液相色谱法测定钴胺素光化动力学装置及方法,属于化工技术领域。装置由原料罐(1)、原料罐温度控制系统(2)、原料罐出口管线温度控制(3)、原料泵(4)、原料泵出口管线温度控制组(5)、n个进料阀、n个出料阀、n个微通道反应器、在线六通阀(16)、高效液相色谱(17)、废液收集系统(18)、光源发生装置(19)及光漫射系统(25)组成。本发明具有结构简单、操作方便、实验效率高、能快速测定钴胺素光化动力学等优点。(The invention discloses a device and a method for determining photochemical kinetics of cobalamin by liquid chromatography, belonging to the technical field of chemical industry. The device comprises a raw material tank (1), a raw material tank temperature control system (2), a raw material tank outlet pipeline temperature control group (3), a raw material pump (4), a raw material pump outlet pipeline temperature control group (5), n feed valves, n discharge valves, n micro-channel reactors, an online six-way valve (16), a high performance liquid chromatography (17), a waste liquid collecting system (18), a light source generating device (19) and a light diffusion system (25). The invention has the advantages of simple structure, convenient operation, high experimental efficiency, capability of rapidly measuring the photochemical kinetics of the cobalamins and the like.)

1. The method for determining the photochemical kinetics of the cobalamin by using the liquid chromatography is characterized by comprising the following steps:

(a) adding a cyanocobalamin aqueous solution or a methylcobalamin aqueous solution or an adenosylcobalamin aqueous solution with a certain concentration into a raw material tank (1), controlling the temperature TC01 of a raw material tank temperature control system (2), the temperature TC02 of a raw material tank outlet pipeline temperature control system (3) and the temperature TC03 of a raw material pump outlet pipeline temperature control group (5) to be T₀Turning on the light source generating device (19) until the temperature stabilizes;

(b) opening the No. 1 feed valve (6) and the No. 1 discharge valve (7), closing the rest feed valves and discharge valves, opening the raw material pump (4), and performing high performance liquid chromatography (1)7) On-line sampling analysis of the composition X of the raw material and the product in the material after passing through a No. 1 micro-channel reactor (20)₁、Y₁After the test is finished, the raw material pump (4) is closed, and the No. 1 feeding valve (6) and the No. 1 discharging valve (7) are closed;

(c) opening a No. 2 feed valve (8) and a No. 2 discharge valve (9), closing the rest feed valves and discharge valves, opening a raw material pump (4), and analyzing the composition X of raw materials and products in the materials after flowing through a No. 2 microchannel reactor (21) through high performance liquid chromatography (17) online sampling₂、Y₂After the test is finished, the raw material pump (4) is closed, and the No. 2 feeding valve (8) and the No. 2 discharging valve (9) are closed;

(d) sequentially testing the composition of the material after passing through the remaining microchannel reactor according to the method in step (b) to obtain the composition at temperature T₀Then, the products corresponding to different light source positions form a variation curve;

(e) the temperature TC01 of the temperature control system (2) of the raw material tank, the temperature TC02 of the temperature control (3) of the outlet pipeline of the raw material tank and the temperature TC03 of the temperature control group (5) of the outlet pipeline of the raw material pump are all T₁After the temperature is stable, the composition of the materials after the raw materials pass through No. 1-n microchannel reactors is tested in sequence according to the method in the step (b), and the temperature T can be obtained₁Then, the products corresponding to different light source positions form a variation curve;

(f) obtaining a product composition change curve corresponding to different light source positions at a series of temperatures according to the method in the step (e);

(g) two kinetic curves can be obtained through the test data of the steps (b) to (f): different light source position-product composition change curves corresponding to the same temperature and different temperature-product composition change curves corresponding to the same light source position.

2. The method of claim 1 for determining photochemical kinetics of cobalamin by liquid chromatography, wherein: the temperature TC01 range of the raw material tank temperature control system (2) in the step (a) is as follows: -5 ℃ to 90 ℃, the temperature TC02 of the head tank outlet line temperature control (3) ranges: -5 ℃ to 90 ℃, the temperature TC03 of the raw material pump outlet line temperature control group (5) being in the range: -5 ℃ to 90 ℃ while satisfying: TC01 ═ TC02 ═ TC 03.

3. The device for determining the photochemical kinetics of cobalamin by using the liquid chromatography comprises a raw material tank (1), a raw material tank temperature control system (2), a raw material tank outlet pipeline temperature control group (3), a raw material pump (4), a raw material pump outlet pipeline temperature control group (5), a feed valve (6) No. 1, a discharge valve (7) No. 1, a feed valve (8) No. 2, a discharge valve (9) No. 2, a feed valve (10) No. 3, a discharge valve (11) No. 3, a feed valve No. n-1 (12), a discharge valve No. n-1 (13), a feed valve No. n (14), a discharge valve No. n (15), an online six-way valve (16), a high performance liquid chromatograph (17), a waste liquid collecting system (18), a light source generating device (19), a microchannel reactor No. 1 (20), a microchannel reactor No. 2 (21), a microchannel reactor No. 3 (22), a microchannel reactor No. n-1 (23), A number n microchannel reactor (24) and a light diffusing system (25), characterized by: raw material tank (1) link to each other through pipeline and former feed pump (4), the outside of raw material tank (1) is adhered to and has former feed tank temperature control system (2), the outside of the pipeline that links to each other between former feed tank (1) and former feed pump (4) is adhered to and has former feed tank export pipeline temperature control (3), the export of former feed pump (4) and the entry of 1 number feed valve (6), the entry of 2 number feed valve (8), the entry of 3 number feed valve (10), the entry of n-1 number feed valve (12), the entry of n number feed valve (14) pass through the pipeline and link to each other and the pipeline outside that is connected is adhered to and has raw material pump export pipeline temperature control group (5), the export of 1 number feed valve (6) pass through the pipeline and link to each other with 1 number microchannel reactor (20), 1 number discharge valve (7), the 1 end of online six-way valve (16) in proper order, the export of 2 number feed valve (8) pass through the pipeline and link to each other with 2 number microchannel reactor (21) in proper order, The discharge valve 2 (9) and the end 1 of the online six-way valve (16) are connected, the outlet of the feed valve 3 (10) is sequentially connected with the ends 1 of the microchannel reactor 3 (22), the discharge valve 3 (11) and the online six-way valve (16) through pipelines, the outlet of the feed valve n-1 (12) is sequentially connected with the ends 1 of the microchannel reactor n-1 (23), the discharge valve n-1 (13) and the online six-way valve (16) through pipelines, the outlet of the feed valve n (14) is sequentially connected with the ends 1 of the microchannel reactor n (24), the discharge valve n (15) and the online six-way valve (16) through pipelines, the end 2 of the online six-way valve (16) is connected with the waste liquid collecting system (18) through pipelines, and the ends 3 and 4 of the online six-way valve (16) are connected with the high performance liquid chromatography (17) through pipelines, the number 1 micro-channel reactor (20), the number 2 micro-channel reactor (21), the number 3 micro-channel reactor (22), the number n-1 micro-channel reactor (23) and the number n micro-channel reactor (24) are all arranged in a light diffusion system (25), and a light source generating device (19) is arranged above the light diffusion system (25).

4. The apparatus for determining the photochemical kinetics of cobalamin by liquid chromatography as claimed in claim 3, wherein: the light source in the light source generating device (19) can be a single point light source or a single line light source or a combination of a plurality of point light sources arranged according to a certain rule or a combination of a plurality of line light sources arranged according to a certain rule or a combination of a plurality of point light sources and line light sources arranged according to a certain rule.

5. The apparatus for determining the photochemical kinetics of cobalamin by liquid chromatography as claimed in claim 3, wherein: the number 1 micro-channel reactor (20), the number 2 micro-channel reactor (21), the number 3 micro-channel reactor (22), the number n-1 micro-channel reactor (23) and the number n micro-channel reactor (24) are all made of quartz glass, the outer diameter is phi 3mm or phi 4mm, and the wall thickness is 1 mm.

6. The apparatus for determining the photochemical kinetics of cobalamin by liquid chromatography as claimed in claim 3, wherein: the inner wall of the light diffusion system (25) is provided with a light reflection layer, the light diffusion system (25) is provided with n groups of round holes with the diameter being r based on axial symmetry, the connecting line of the circle centers of the round holes is vertical to the central axis of the light diffusion system (25), the height of the light diffusion system (25) is H, the circle center distance between the adjacent round holes is H, and the range of r is as follows: 4 mm-6 mm, and the relationship among H, H and r satisfies: h is more than or equal to 1.5r and less than or equal to H/(n + 1).

The technical field is as follows:

the invention discloses a device and a method for measuring photochemical kinetics of cobalamin by liquid chromatography, belongs to the technical field of chemical industry, and is suitable for researching a kinetics curve of cyanocobalamin, methylcobalamin or adenosylcobalamin generated by photochemical reaction of hydroxycobalamin.

Background art:

photochemical reactions refer to chemical reactions initiated by the absorption of a photon by an atom, molecule, radical or ion. Common photochemical reactions include photo-oxidation, photo-reduction, photo-polymerization, and photo-substitution. For example: cyanocobalamin, methylcobalamin or desoxyadenosylcobalamin are subjected to light irradiation to generate hydroxycobalamin, which belongs to the light substitution reaction. The reactions have the characteristics of mild reaction, high recovery rate, good selectivity and the like.

The photochemical reaction process is relatively complicated. First, photochemistry includes a series of complex photophysical and photochemical phenomena such as photoinduced electron transfer, excited proton transfer, potential energy plane crossing, photoisomerization, photodissociation, internal conversion, intersystem crossing and the like; second, excited molecules have high energy but short lifetimes and often lose energy to return to the ground state before chemical reaction, so whether a photochemical reaction occurs depends on the relative rates of the excited chemical reaction process and the energy decay process; third, the photochemical and photophysical changes of the excited molecule are a relatively competing process and a process that coexist with each other.

The reaction rate of the photochemical method for preparing the hydroxycobalamin is related to factors such as wavelength of absorbed light, light quantum flux, temperature and the like, and the research of a related kinetic curve has profound significance for guiding the process research of preparing the hydroxycobalamin by the photochemical method.

The invention content is as follows:

the invention aims to provide an experimental device and an experimental method which have simple structure, convenient operation and high experimental efficiency and can rapidly measure the photochemical kinetics of cobalamin by liquid chromatography.

The invention provides a method for determining photochemical kinetics of cobalamin by liquid chromatography, which comprises the following steps:

(a) adding a cyanocobalamin aqueous solution or a methylcobalamin aqueous solution or an adenosylcobalamin aqueous solution with a certain concentration into a raw material tank (1), controlling the temperature TC01 of a raw material tank temperature control system (2), the temperature TC02 of a raw material tank outlet pipeline temperature control system (3) and the temperature TC03 of a raw material pump outlet pipeline temperature control group (5) to be T₀Turning on the light source generating device (19) until the temperature stabilizes;

(b) opening a feed valve (6) No. 1 and a discharge valve (7) No. 1, closing the rest feed valves and discharge valves, opening a raw material pump (4), and analyzing the composition X of raw materials and products in the materials after flowing through a microchannel reactor (20) No. 1 by high performance liquid chromatography (17) online sampling₁、Y₁After the test is finished, the raw material pump (4) is closed, and the No. 1 feeding valve (6) and the No. 1 discharging valve (7) are closed;

(c) opening No. 2 feed valve (8) and No. 2 bleeder valve (9), closing the rest feed valves and bleeder valves, opening raw material pump (4), and passing through high performance liquid chromatography (1)7) On-line sampling analysis of the composition X of the raw material and the product in the material after passing through a No. 2 micro-channel reactor (21)₂、Y₂After the test is finished, the raw material pump (4) is closed, and the No. 2 feeding valve (8) and the No. 2 discharging valve (9) are closed;

(d) sequentially testing the composition of the material after passing through the remaining microchannel reactor according to the method in step (b) to obtain the composition at temperature T₀Then, the products corresponding to different light source positions form a variation curve;

(e) the temperature TC01 of the temperature control system (2) of the raw material tank, the temperature TC02 of the temperature control (3) of the outlet pipeline of the raw material tank and the temperature TC03 of the temperature control group (5) of the outlet pipeline of the raw material pump are all T₁After the temperature is stable, the composition of the materials after the raw materials pass through No. 1-n microchannel reactors is tested in sequence according to the method in the step (b), and the temperature T can be obtained₁Then, the products corresponding to different light source positions form a variation curve;

(f) obtaining a product composition change curve corresponding to different light source positions at a series of temperatures according to the method in the step (e);

(g) two kinetic curves can be obtained through the test data of the steps (b) to (f): different light source position-product composition change curves corresponding to the same temperature and different temperature-product composition change curves corresponding to the same light source position.

The method for determining the photochemical kinetics of the cobalamin by the liquid chromatography is further characterized in that: the temperature TC01 range of the raw material tank temperature control system (2) in the step (a) is as follows: -5 ℃ to 90 ℃, the temperature TC02 of the head tank outlet line temperature control (3) ranges: -5 ℃ to 90 ℃, the temperature TC03 of the raw material pump outlet line temperature control group (5) being in the range: -5 ℃ to 90 ℃ while satisfying: TC01 ═ TC02 ═ TC 03.

The invention also provides a device for determining the photochemical kinetics of cobalamin by using the liquid chromatography, which comprises a raw material tank (1), a raw material tank temperature control system (2), a raw material tank outlet pipeline temperature control group (3), a raw material pump (4), a raw material pump outlet pipeline temperature control group (5), a No. 1 feed valve (6), a No. 1 discharge valve (7), a No. 2 feed valve (8), a No. 2 discharge valve (9), a No. 3 feed valve (10), a No. 3 discharge valve (11), an n-1 feed valve (12), an n-1 discharge valve (13), an N feed valve (14), an N discharge valve (15), an online six-way valve (16), a high performance liquid chromatography (17), a waste liquid collection system (18), a light source generation device (19), a No. 1 microchannel reactor (20), a No. 2 microchannel reactor (21), a No. 3 microchannel reactor (22), No. n-1 microchannel reactor (23), No. n microchannel reactor (24) and light diffusion system (25), characterized in that: raw material tank (1) link to each other through pipeline and former feed pump (4), the outside of raw material tank (1) is adhered to and has former feed tank temperature control system (2), the outside of the pipeline that links to each other between former feed tank (1) and former feed pump (4) is adhered to and has former feed tank export pipeline temperature control (3), the export of former feed pump (4) and the entry of 1 number feed valve (6), the entry of 2 number feed valve (8), the entry of 3 number feed valve (10), the entry of n-1 number feed valve (12), the entry of n number feed valve (14) pass through the pipeline and link to each other and the pipeline outside that is connected is adhered to and has raw material pump export pipeline temperature control group (5), the export of 1 number feed valve (6) pass through the pipeline and link to each other with 1 number microchannel reactor (20), 1 number discharge valve (7), the 1 end of online six-way valve (16) in proper order, the export of 2 number feed valve (8) pass through the pipeline and link to each other with 2 number microchannel reactor (21) in proper order, The discharge valve 2 (9) and the end 1 of the online six-way valve (16) are connected, the outlet of the feed valve 3 (10) is sequentially connected with the ends 1 of the microchannel reactor 3 (22), the discharge valve 3 (11) and the online six-way valve (16) through pipelines, the outlet of the feed valve n-1 (12) is sequentially connected with the ends 1 of the microchannel reactor n-1 (23), the discharge valve n-1 (13) and the online six-way valve (16) through pipelines, the outlet of the feed valve n (14) is sequentially connected with the ends 1 of the microchannel reactor n (24), the discharge valve n (15) and the online six-way valve (16) through pipelines, the end 2 of the online six-way valve (16) is connected with the waste liquid collecting system (18) through pipelines, and the ends 3 and 4 of the online six-way valve (16) are connected with the high performance liquid chromatography (17) through pipelines, the number 1 micro-channel reactor (20), the number 2 micro-channel reactor (21), the number 3 micro-channel reactor (22), the number n-1 micro-channel reactor (23) and the number n micro-channel reactor (24) are all arranged in a light diffusion system (25), and a light source generating device (19) is arranged above the light diffusion system (25).

The device for determining the photochemical kinetics of the cobalamin by the liquid chromatography is further characterized in that: the light source in the light source generating device (19) can be a single point light source or a single line light source or a combination of a plurality of point light sources arranged according to a certain rule or a combination of a plurality of line light sources arranged according to a certain rule or a combination of a plurality of point light sources and line light sources arranged according to a certain rule.

The device for determining the photochemical kinetics of the cobalamin by the liquid chromatography is further characterized in that: the number 1 micro-channel reactor (20), the number 2 micro-channel reactor (21), the number 3 micro-channel reactor (22), the number n-1 micro-channel reactor (23) and the number n micro-channel reactor (24) are all made of quartz glass, the outer diameter is phi 3mm or phi 4mm, and the wall thickness is 1 mm.

The device for determining the photochemical kinetics of the cobalamin by the liquid chromatography is further characterized in that: the inner wall of the light diffusion system (25) is provided with a light reflection layer, the light diffusion system (25) is provided with n groups of round holes with the diameter being r based on axial symmetry, the connecting line of the circle centers of the round holes is vertical to the central axis of the light diffusion system (25), the height of the light diffusion system (25) is H, the circle center distance between the adjacent round holes is H, and the range of r is as follows: 4 mm-6 mm, and the relationship among H, H and r satisfies: h is more than or equal to 1.5r and less than or equal to H/(n + 1).

The device and the method for determining photochemical kinetics of cobalamin by using the liquid chromatography have the following advantages that: the device has the advantages of simple structure, compact layout, convenient operation and simple experimental method, and the product composition can be analyzed on line through the high performance liquid chromatography (17) only by changing the temperature, the flow, the light source, the valve switches and the like after the raw materials are thrown, so that the one-time feeding rapid analysis is realized, and the experimental efficiency and the automation degree are improved.

Description of the drawings:

FIG. 1 is a schematic diagram of a photochemical kinetic device for determining cobalamin by liquid chromatography.

Fig. 2 is a schematic view of the structure of the light diffusion system (25).

The specific implementation mode is as follows:

in order to make the present invention more comprehensible, preferred embodiments are described in detail below with reference to the accompanying drawings, but the present invention is not limited thereto.

As shown in figure 1, the device for determining the photochemical kinetics of cobalamin by using the liquid chromatography comprises a raw material tank (1), a raw material tank temperature control system (2), a raw material tank outlet pipeline temperature control (3), a raw material pump (4), a raw material pump outlet pipeline temperature control group (5), a No. 1 feed valve (6), a No. 1 discharge valve (7), a No. 2 feed valve (8), a No. 2 discharge valve (9), a No. 3 feed valve (10), a No. 3 discharge valve (11), an n-1 feed valve (12), an n-1 discharge valve (13), an N feed valve (14), an N discharge valve (15), an online six-way valve (16), a high performance liquid chromatography (17), a waste liquid collection system (18), a light source generation device (19), a No. 1 microchannel reactor (20), a No. 2 microchannel reactor (21), a No. 3 microchannel reactor (22), n-1 microchannel reactor (23), n microchannel reactor (24) and light diffusion system (25), raw material tank (1) links to each other with feedstock pump (4) through the pipeline, raw material tank temperature control system (2) are adhered to the outside of raw material tank (1), raw material tank export pipeline temperature control (3) are adhered to the outside of the pipeline that links to each other between feedstock tank (1) and feedstock pump (4), the export of feedstock pump (4) and the entry of 1 feed valve (6), the entry of 2 feed valve (8), the entry of 3 feed valve (10), the entry of n-1 feed valve (12), the entry of n feed valve (14) links to each other through the pipeline and the pipeline outside of being connected is adhered to raw material pump export pipeline temperature control group (5), the export of 1 feed valve (6) is in proper order through the pipeline with 1 microchannel reactor (20), The No. 1 discharge valve (7) and the end 1 of the online six-way valve (16) are connected, the outlet of the No. 2 feed valve (8) is sequentially connected with the end 1 of the No. 2 microchannel reactor (21), the No. 2 discharge valve (9) and the online six-way valve (16) through pipelines, the outlet of the No. 3 feed valve (10) is sequentially connected with the end 1 of the No. 3 microchannel reactor (22), the No. 3 discharge valve (11) and the online six-way valve (16) through pipelines, the outlet of the No. n-1 feed valve (12) is sequentially connected with the end 1 of the No. n-1 microchannel reactor (23), the No. n-1 discharge valve (13) and the online six-way valve (16) through pipelines, the outlet of the No. n feed valve (14) is sequentially connected with the end 1 of the No. n microchannel reactor (24), the No. n discharge valve (15) and the online six-way valve (16) through pipelines, and the end 2 of the online six-way valve (16) is connected with the waste liquid collecting system (18) through a pipeline, the 3 ends and 4 ends of the online six-way valve (16) are connected with a high performance liquid chromatograph (17) through pipelines, a No. 1 micro-channel reactor (20), a No. 2 micro-channel reactor (21), a No. 3 micro-channel reactor (22), an n-1 micro-channel reactor (23) and an n micro-channel reactor (24) are all placed in a light diffusion system (25), the materials of the micro-channel reactors are quartz glass, the outer diameter size is phi 3mm or phi 4mm, the wall thickness is 1mm, a light source generating device (19) is arranged above the light diffusion system (25), a light source in the light source generating device (19) can be a single point light source or a single line light source or a plurality of point light source combinations arranged according to a certain rule or a plurality of line light source combinations arranged according to a certain rule or a plurality of point light sources and line light source combinations arranged according to a certain rule, and a light reflection layer is arranged on the inner wall of the light diffusion system (25), the light diffusion system (25) is provided with n groups of round holes with the diameter of r based on axial symmetry, the connecting line of the circle centers of the round holes is vertical to the central axis of the light diffusion system (25), the height of the light diffusion system (25) is H, the distance between the centers of the adjacent round holes is H, and the range of r is as follows: 4 mm-6 mm, and the relationship among H, H and r satisfies: h is more than or equal to 1.5r and less than or equal to H/(n + 1).

The structure of the device for measuring the photochemical kinetics of the cobalamin is determined according to the liquid chromatography, and the method for measuring the photochemical kinetics of the cobalamin by the liquid chromatography is as follows:

(a) adding a cyanocobalamin aqueous solution or a methylcobalamin aqueous solution or an adenosylcobalamin aqueous solution with a certain concentration into a raw material tank (1), controlling the temperature TC01 of a raw material tank temperature control system (2), the temperature TC02 of a raw material tank outlet pipeline temperature control system (3) and the temperature TC03 of a raw material pump outlet pipeline temperature control group (5) to be T₀，T₀The range is as follows: -5 ℃ to 90 ℃ until the temperature is stable, turning on the light source generating means (19);

(b) opening a feed valve (6) No. 1 and a discharge valve (7) No. 1, closing the rest feed valves and discharge valves, opening a raw material pump (4), and analyzing the composition X of raw materials and products in the materials after flowing through a microchannel reactor (20) No. 1 by high performance liquid chromatography (17) online sampling₁、Y₁After the test is finished, the raw material pump (4) is closed, and the No. 1 feeding valve (6) and the No. 1 discharging valve (7) are closed;

(c) opening the No. 2 feed valve (8) and the No. 2 discharge valve (9), closing the rest feed valves and discharge valves, opening the raw material pump (4), and taking out the raw material on line through the high performance liquid chromatography (17)Sample analysis of the composition X of the feed and product after passing through a No. 2 microchannel reactor (21)₂、Y₂After the test is finished, the raw material pump (4) is closed, and the No. 2 feeding valve (8) and the No. 2 discharging valve (9) are closed;

(d) sequentially testing the composition of the material after passing through the remaining microchannel reactor according to the method in step (b) to obtain the composition at temperature T₀Then, the products corresponding to different light source positions form a variation curve;

(e) the temperature TC01 of the temperature control system (2) of the raw material tank, the temperature TC02 of the temperature control (3) of the outlet pipeline of the raw material tank and the temperature TC03 of the temperature control group (5) of the outlet pipeline of the raw material pump are all T₁，T₁The range is as follows: the temperature is between 5 ℃ below zero and 90 ℃ until the temperature is stable, the composition of the materials after the raw materials pass through No. 1 to n microchannel reactors is tested in sequence according to the method in the step (b), and the temperature T can be obtained₁Then, the products corresponding to different light source positions form a variation curve;

(f) obtaining a product composition change curve corresponding to different light source positions at a series of temperatures according to the method in the step (e);

(g) two kinetic curves can be obtained through the test data of the steps (b) to (f): different light source position-product composition change curves corresponding to the same temperature and different temperature-product composition change curves corresponding to the same light source position.