A method for online reaction and mass spectrometric in situ sampling analysis

Abstract

The application discloses a method for online reaction and mass spectrum in-situ sampling analysis, and belongs to the technical field of chemical analysis. The method is used for sampling and analyzing a sample in real time without destroying a reaction environment or prematurely terminating the reaction, avoids destroying the sample by an external environment, improves the accuracy of experimental results, and effectively reduces labor cost.

Description

Technical Field

[0001] This invention belongs to the field of chemical analysis technology, specifically relating to a method for online reaction and in-situ mass spectrometry analysis. Background Technology

[0002] In sulfinic acid-bridged bisferric complexes, the iron atom exists in multiple oxidation states, and these complexes can interconvert between different oxidation states. Under certain conditions, they can accept or lose electrons, participating in redox reactions. They can act as catalysts in various organic reactions, such as oxidation, reduction, and coupling reactions. Their unique structure and electronic properties can activate substrate molecules, lower reaction activation energies, and improve reaction selectivity and efficiency. In the biomedical field, due to their structural similarity to some metalloproteins and enzymes in living organisms, sulfinic acid-bridged bisferric complexes have potential applications. For example, they could serve as design templates for drug molecules, used to develop novel antibacterial, antiviral, or antitumor drugs; or as bioimaging reagents, used to detect the concentration of metal ions or the distribution of specific biomolecules in living organisms.

[0003] Sulfinate-bridged diferric complexes are often prepared by the oxidation of biomimetic binuclear iron-sulfur clusters. However, biomimetic binuclear iron-sulfur clusters are sensitive to oxygen and are easily oxidized in the air. Sampling often disrupts the reaction environment or terminates the reaction. Therefore, conventional sampling methods cannot be used for testing and characterization. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a method for online reaction and in-situ mass spectrometry analysis, which allows for real-time sampling and analysis of samples without disrupting the reaction environment or prematurely terminating the reaction.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows: A method for online reaction and in-situ mass spectrometry sample injection analysis, comprising the following steps:

[0006]

[0007] S1. First, draw nitrogen gas into feed tubes 1 and 2 of the feeding device through a nitrogen protection bottle. Then, place feed tube 1 into CH2Cl2 and feed tube 2 into the CH2Cl2 solution of compound 1. Start the feeding device. The ratio of feed in feed tube 1 to feed tube 2 is 9:1.

[0008] S2. The feeding device delivers CH2Cl2 and the CH2Cl2 solution of compound 1 to the reaction device through the liquid inlet pipe, and mixes them evenly; sampling and analysis are performed, and the peak m / z of compound 1 is found to be 538.0.

[0009] S3. The reaction three-way valve connects the oxygen outlet pipe and the gas inlet pipe; the oxygen in the oxygen source cylinder enters the reaction device through the oxygen outlet pipe, the reaction three-way valve, and the gas inlet pipe in sequence. When the pressure sensor monitors that the pressure inside the device reaches 1 atm, stirring begins and the reaction timer starts.

[0010] S4. Set the sampling volume to 0.1 mL, perform sampling and analysis, and obtain the peak m / z of compound 2 as 570.0; set different sampling and analysis times to monitor the reaction;

[0011] The sampling process in steps S2 and S4 is as follows: the injection three-way valve is connected to the syringe connecting tube and the six-way valve and three-way valve connecting tube; the injection six-way valve is connected to the six-way valve and three-way valve connecting tube and the filter and injection connecting tube; the syringe is started to draw in suction, a negative pressure is formed in the tubing, the sample in the reaction device is drawn out, the sample is filtered through the liquid outlet tube and then enters the syringe in sequence through the filter and injection connecting tube, the injection six-way valve, the six-way valve and three-way valve connecting tube, the injection three-way valve, and the syringe connecting tube;

[0012] The three-way injection valve connects to the syringe connecting tube and the six-way valve and three-way valve connecting tube. The six-way injection valve connects to the six-way valve and three-way valve connecting tube and the injection and analysis connecting tube. The syringe pushes the sample through the syringe connecting tube, the six-way valve and three-way valve connecting tube, and the injection and analysis connecting tube in sequence into the analytical device for analysis.

[0013] Compared to existing technologies, this invention offers the following advantages: The method allows for real-time sampling and analysis of samples without disrupting the reaction environment or prematurely terminating the reaction, avoiding external environmental damage, improving the accuracy of experimental results, and effectively reducing labor costs. It also effectively reduces personnel contact with chemical reagents, minimizing the risk of health harm caused by unexpected accidents or prolonged exposure during experiments. Furthermore, this invention avoids reaction uncertainties resulting from raw material oxidation during sampling, effectively controls the reaction endpoint, improves the accuracy of experimental monitoring, and facilitates the study of reaction mechanisms. Attached Figure Description

[0014] Figure 1 This is a schematic diagram of the online reaction and analysis system.

[0015] Figure 2 These are graphs showing the analysis results obtained from sampling at different times.

[0016] In the diagram: 1. Auxiliary sample injection device; 2. Nitrogen protection bottle; 3. Feeding device; 4. Sample injection device; 5. Analytical device; 6. Cleaning solution bottle; 7. Reaction device; 8. Filtering device; 9. Nitrogen gas source bottle; 9a. Oxygen gas source bottle; 10. Auxiliary sample injection line; 11. Feeding line; 12. Syringe connection line; 13. Valve and cleaning bottle connection line; 14. Six-way valve and three-way valve connection line; 15. Liquid inlet pipe; 16. Filtering and sample injection connection line; 17. Sample injection and analysis connection line; 18. Liquid outlet pipe; 19. Gas inlet pipe; 20. Nitrogen outlet pipe; 21. Oxygen outlet pipe; 22. Analytical device outlet pipe; 23. Reaction three-way valve; 24. Sample injection three-way valve; 25. Sample injection six-way valve; 26. Syringe. Detailed Implementation

[0017] The technical solution of the present invention will be clearly described below with reference to the accompanying drawings. Obviously, the described embodiments are not all embodiments of the present invention. All other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.

[0018] It should be noted that, unless otherwise specifically stated, the relative arrangement and numerical expressions of the components and steps described in these embodiments should not be construed as limiting the scope of the invention.

[0019] The following description of exemplary embodiments is merely illustrative and is not intended to limit the invention or its application or use in any way. Techniques, methods, and apparatus known to those skilled in the art may not be discussed in detail herein, but where applicable, such techniques, methods, and apparatus should be considered part of this specification.

[0020] Figure 1 An online reaction and analysis system for the oxidation of biomimetic binuclear iron-sulfur clusters is shown. The oxidation reaction includes the following steps:

[0021]

[0022] S1. First, draw nitrogen gas into feed tubes 1 and 2 of feed device 3 through nitrogen protection bottle 2, then put feed tube 1 into CH2Cl2 and feed tube 2 into the CH2Cl2 solution of compound 1; start feed device 3, and the ratio of feed in feed tube 1 to feed tube 2 is 9:1.

[0023] S2, the feeding device 3 delivers CH2Cl2 and the CH2Cl2 solution of compound 1 to the reaction device 7 through the liquid inlet pipe 15, and mixes them evenly; sampling and analysis are performed, and the peak m / z of compound 1 is found to be 538.0;

[0024] S3, the reaction three-way valve 23 connects the oxygen outlet pipe 21 and the gas inlet pipe 19; the oxygen in the oxygen source cylinder 9a enters the reaction device 7 through the oxygen outlet pipe 21, the reaction three-way valve 23, and the gas inlet pipe 19 in sequence. When the pressure sensor monitors that the pressure in the device reaches 1 atm, stirring begins and the reaction timer starts.

[0025] S4. Set the sampling volume to 0.1 mL, perform sampling and analysis, and obtain the peak m / z of compound 2 as 570.0; set different sampling and analysis times to monitor the reaction;

[0026] The sampling process is as follows: the injection three-way valve 24 is connected to the syringe connecting tube 12 and the six-way valve and three-way valve connecting tube 14; the injection six-way valve 25 is connected to the six-way valve and three-way valve connecting tube 14 and the filter and injection connecting tube 16; the syringe 26 is started to draw in the sample, a negative pressure is formed in the tubing, and the sample in the reaction device 7 is drawn out. The sample is filtered by the filter device 8 through the liquid outlet tube 18, and then enters the syringe 26 in sequence through the filter and injection connecting tube 16, the injection six-way valve 25, the six-way valve and three-way valve connecting tube 14, the injection three-way valve 24, and the syringe connecting tube 12.

[0027] The injection three-way valve 24 connects to the syringe connecting tube 12 and the six-way valve and three-way valve connecting tube 14. The injection six-way valve 25 connects to the six-way valve and three-way valve connecting tube 14 and the injection and analysis connecting tube 17. The syringe 26 pushes the sample through the syringe connecting tube 12, the six-way valve and three-way valve connecting tube 14, and the injection and analysis connecting tube 17 into the analysis device 5 for analysis.

[0028] The feeding device 3 is a quaternary low-pressure pump with a settable flow rate range of 0~10 mL / min. It has four feed lines 11, which can simultaneously draw up the chemical reagents required for four different chemical reactions. The feed lines 11 are made of PTFE, which has good corrosion resistance to acids, alkalis, and organic reagents. The other end of the feed line 11 is placed in a nitrogen protection bottle 2. The feeding device 3 has one outlet line - a liquid inlet line 15, made of the same material as the feed lines 11, which is connected to the liquid inlet of the reaction apparatus 7, enabling automatic feeding according to the reaction requirements.

[0029] The reaction device 7 is designed for explosion-proof operation. It consists of an inner frame and an outer frame. A Peltier element is placed between the inner and outer frames to provide heating and cooling for the reactor body. Insulation cotton is placed between the inner and outer frames, avoiding the Peltier element, to prevent heat loss from the device. A control panel and temperature display are located on the outer surface of the outer frame. The control panel allows for setting the functional parameters of the feeding device, reaction device, sample injection device, and cleaning device to achieve automated reaction control. The temperature display shows the current reaction temperature.

[0030] A 2-position 3-way electromagnetic switching valve—a reaction three-way valve 23—is placed on the outer surface of the outer frame of the reaction device 7. One port of the electromagnetic switching valve is connected to the nitrogen outlet pipe 21, another port is connected to the nitrogen outlet pipe 20, and the remaining port serves as the gas outlet of the electromagnetic switching valve, connected to the reaction device cover via the gas inlet pipe 19, enabling automatic switching between different gas inputs. The reaction device cover is arc-shaped and has a sensor on it to monitor liquid level changes and the pressure and temperature inside the reactor.

[0031] The inner liner of the reaction apparatus is made of quartz and is placed within the inner frame of the apparatus. It can be removed, cleaned, and reused after the reaction. A magnetic stir bar is placed at the bottom for stirring during the reaction.

[0032] The inlet of the filter device 8 is connected to the liquid outlet pipe 18 on the reaction device 7, and the outlet is connected to the sample injection device 4 through the filter and sample injection connection pipe 16. Its function is to filter the sample solution drawn by the sample injection device as needed to avoid contaminating the sample injection device and the detection device.

[0033] The sample injection device 4 has its inlet connected to the outlet of the filter device 8. It comprises a 2-position 6-way solenoid switching valve-sample injection six-way valve 25, a 2-position 3-way solenoid valve-sample injection three-way valve 24, and a syringe 26. The 2-position 6-way solenoid switching valve-sample injection six-way valve 25 has two ports connected to a quantitative loop, one port connected to the analyzer device 5 via a sample injection and analysis connection tube, one port connected to the filter and sample injection connection tube 16, one port connected to the auxiliary sample injection device 1 via an auxiliary sample injection line 10, and one port connected to the 2-position 3-way solenoid valve-sample injection three-way valve 24 via a six-way valve and three-way valve connection tube 14. The remaining two ports of the 2-position 3-way solenoid valve-sample injection three-way valve 24 are connected as follows: one port is connected to the cleaning solution bottle 6 via a valve and washing solution bottle connection tube 13, providing washing solution for syringe 26 during cleaning; the other port is connected to syringe 26 via syringe connection tube 12. During sample injection, the three-way valve 24 switches to the syringe connection channel to draw up the sample for injection; during cleaning, the three-way valve 24 switches to the cleaning solution channel to draw up the cleaning solution to clean the injection device tubing.

[0034] The analytical device 5 is typically a gas chromatograph (GC), a mass spectrometer (MS), or a gas chromatography-mass spectrometer-mass spectrometer (GC-MS). By setting the instrument's functional parameters according to the sample properties, the sample can be analyzed automatically.

[0035] The auxiliary sample injection device 1 is a single pump connected to the sample injection device through pipeline 10. It draws up specific reagents, such as acetonitrile and dichloromethane, and transports the sample to the analytical device.

[0036] Example 1

[0037] Based on the reaction conditions, the reaction and product analysis were performed using an automated, multifunctional online reaction and mass spectrometry in-situ sample introduction system provided by this invention.

[0038] 1) First, draw nitrogen into the feed pipes 1 and 2 of the feeding device through the nitrogen protection bottle. Then, put pipe 1 into CH2Cl2 and pipe 2 into the CH2Cl2 solution of biomimetic sulfur-bridged double iron ammonium compound 1. Connect the solenoid valve 1 on the reaction device to the O2 gas cylinder and the solenoid valve 2 to the nitrogen gas cylinder. Connect the gas outlet pipe to the gas inlet pipe on the reactor cover.

[0039] 2) Using the control panel, set the flow rate of the feed device to 5 mL / min, the time to 2.5 min, the channel selection to 1 and 2, the ratio to 9:1, the gas channel of the reaction device to 1 (O2), the pressure to 1 atm, the temperature to room temperature, and the reaction time to 24 h. After setting these parameters, run the system.

[0040] 3) The feeding device simultaneously delivers CH2Cl2 and the CH2Cl2 solution of compound 1 into the inner liner of the reaction device through pipeline in a certain proportion. After stirring evenly, a sample is taken and tested, and the raw material peak m / z 538.0 is obtained.

[0041] 4) Switch the solenoid valve on the reaction apparatus to position 1 via the control panel. Inject O2 into the reaction apparatus through the pipeline. Once the pressure sensor detects that the pressure inside the apparatus reaches 1 atm, and the reaction parameters are ready, turn on the stirring function and start the reaction timing. Set the sampling volume to 0.1 mL, and inject samples into the MS at different times (see...). Figure 2 The product peak at m / z is 570.0, and the reaction endpoint is monitored.

[0042] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for online reaction and in-situ mass spectrometry injection analysis, characterized in that, The method includes the following steps: S1. First, draw nitrogen gas into feed tubes 1 and 2 of feed device (3) through nitrogen protection bottle (2), then put feed tube 1 into CH2Cl2 and feed tube 2 into CH2Cl2 solution of compound 1; start feed device (3), the ratio of feed in feed tube 1 to feed tube 2 is 9:1; S2, The feeding device (3) transports CH2Cl2 and the CH2Cl2 solution of compound 1 to the reaction device (7) through the liquid inlet pipe (15) and mixes them evenly; samples are taken and analyzed to obtain the peak m / z 538.0 of compound 1; The structure of compound 1 is as follows: ; S3, the reaction three-way valve (23) connects the oxygen outlet pipe (21) and the gas inlet pipe (19); the oxygen in the oxygen source cylinder (9a) enters the reaction device (7) through the oxygen outlet pipe (21), the reaction three-way valve (23) and the gas inlet pipe (19) in sequence. When the pressure sensor monitors that the pressure in the device reaches 1 atm, stirring begins and the reaction timer starts. S4. Set the sampling volume to 0.1 mL, perform sampling and analysis, and obtain the peak m / z of compound 2 as 570.0; set different sampling and analysis times to monitor the reaction; The structure of compound 2 is as follows: ; The sampling process in steps S2 and S4 is as follows: the 2-position 3-way solenoid valve-injection 3-way valve (24) connects the syringe connecting tube (12) and the six-way valve and 3-way valve connecting tube (14); the 2-position 6-way solenoid switching valve-injection 6-way valve (25) connects the six-way valve and 3-way valve connecting tube (14) and the filter and injection connecting tube (16). Start the syringe (26) to draw in the sample. The tubing forms a negative pressure and the sample in the reaction device (7) is drawn out. The sample is filtered by the filter device (8) through the liquid outlet pipe (18) and then passes through the filter and injection connection pipe (16), the 2-position 6-way solenoid switching valve-injection 6-way valve (25), the 6-way valve and 3-way valve connection pipe (14), the 2-position 3-way solenoid valve-injection 3-way valve (24), and the syringe connection pipe (12) into the syringe (26). The 2-position 3-way solenoid valve-sample injection 3-way valve (24) connects to the syringe connecting tube (12) and the 6-way valve and 3-way valve connecting tube (14), and the 2-position 6-way solenoid switching valve-sample injection 6-way valve (25) connects to the 6-way valve and 3-way valve connecting tube (14) and the sample injection and analysis connecting tube (17). The syringe (26) pushes the sample through the syringe connecting tube (12), the six-way valve and three-way valve connecting tube (14), and the sample injection and analysis connecting tube (17) into the analysis device (5) for analysis. The inlet of the filtration device is connected to the liquid outlet pipe on the reaction device, and its outlet is connected to the injection device through the filtration and injection connection pipe. The sample solution drawn by the injection device is filtered as needed. The inlet of the injection device is connected to the outlet of the filtration device. The injection device consists of a 2-position 6-way solenoid switching valve (injection six-way valve), a 2-position 3-way solenoid valve (injection three-way valve), and a syringe. Two ports on the 2-position 6-way solenoid switching valve (injection six-way valve) are connected to a quantitative loop; one port is connected to the analytical device via an injection and analysis connection tube; one port is connected to the filter and injection connection tube; one port is connected to the auxiliary injection device via an auxiliary injection line; and one port is connected to the 2-position 3-way solenoid valve (injection three-way valve) via a six-way valve and three-way valve connection tube. The remaining two ports on the 2-position 3-way solenoid valve (injection three-way valve) are connected as follows: one port is connected to the washing solution bottle via a valve and washing solution bottle connection tube; the other port is connected to the syringe via a syringe connection tube. The auxiliary injection device is a single pump connected to the injection device. After drawing up the reagent, it transports the sample to the analytical device.

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