Self-adjusting lipid photochemical reaction system with controllable reaction efficiency and analysis method
By designing a self-regulated lipid photochemical reaction system with controllable reaction efficiency, the problems of low throughput, poor flexibility and poor reagent compatibility in the prior art are solved, and efficient and accurate lipid photochemical reactions are achieved, suitable for large-scale and Micro sample analysis provides strong technical support for biochemical and medicinal chemistry research.
Patent Information
- Application Number
- CN202411234207.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-04
- Publication Date
- 2024-12-10
AI Technical Summary
The existing photochemical reaction technology has problems such as low flux, poor flexibility, low sample processing efficiency, poor reagent compatibility, unstable reaction reagent injection and inability to monitor reaction efficiency in real time in lipid fine structure analysis, which is difficult to meet the large-scale and high Requirements for flux and micro sample analysis.
A self-regulated lipid photochemical reaction system with controllable reaction efficiency is designed, including lipid injection module, reaction reagent injection module, mixer module, photochemical reaction module, reaction efficiency monitoring module and calculation control module, which is controlled through automatic control. and multi-channel design to achieve efficient and accurate lipid photochemical reactions and are compatible with micro sample analysis.
The analysis throughput is improved, and the precise control of reagent ratio, wavelength and other parameters is achieved, the accuracy and reliability of the analysis results are ensured, and scarce or difficult to obtain lipid samples can be effectively analyzed.
Smart Images
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Abstract
Description
Technical Field
[0001] The present application relates to the technical field of photochemical reaction control and mass spectrometry analysis, and in particular to a self-regulating lipid photochemical reaction system with controllable reaction efficiency and an analysis method. Background Art
[0002] In recent years, lipid research has attracted much attention in the field of life sciences. As an indispensable organic molecule in organisms, the importance of lipids is becoming increasingly prominent. Lipids are the main components of cell membranes. They not only provide structural support and protection for cells, but also participate in many biological functions, such as signal transduction, molecular transmission, energy storage, etc. The fine structure of lipids is directly related to the diversity and specificity of their biological functions. For example, the fatty acid chain composition and connection position of lipid molecules can affect the fluidity, stability and permeability of cell membranes. Structural changes of lipids in head groups, fatty acid chains, C=C positions, sn-positions and other dimensions can significantly affect their distribution in organisms, metabolic pathways and interactions with other biological molecules. Moreover, structural changes in multiple dimensions of lipid fine structure will produce a large number of isomeric molecules, making lipidome analysis challenging.
[0003] In lipid analysis, technologies such as liquid chromatography-mass spectrometry (LC-MS) or gas chromatography-mass spectrometry (GC-MS) have become the main means of lipid analysis. Among the many lipid fine structure mass spectrometry analysis technologies, photochemical reaction-tandem mass spectrometry has attracted much attention due to its unique advantages. It can trigger chemical changes within or between molecules through light excitation of specific wavelengths, thereby achieving precise modification and labeling of lipid structures, making it easier for subsequent tandem mass spectrometry to form specific diagnostic ions for structural identification.
[0004] However, the current application of photochemical reactions in lipid fine structure analysis still faces several challenges: most of the existing technical means rely on "pre-column derivatization", which is not only cumbersome to operate, but also often requires preset reaction conditions, limiting its flexibility and versatility; in addition, existing photochemical reaction equipment usually has a low throughput and a low degree of automation, which cannot meet the needs of large-scale, high-throughput analysis; at the same time, these devices are often unable to adjust the reagent ratio and wavelength, and cannot perform real-time monitoring and feedback on samples after photochemical reactions, limiting their flexibility, accuracy and repeatability in the analysis of different lipid samples; finally, with the deepening of biological research, there is an urgent need for structural analysis of trace biological lipid samples, and existing photochemical reaction equipment also has limitations in the analysis of trace samples. Summary of the invention
[0005] In order to solve the above technical problems, the present application provides a self-regulating lipid photochemical reaction system and an analysis method with controllable reaction efficiency.
[0006] The present application provides a self-regulating lipid photochemical reaction system with controllable reaction efficiency, comprising the following modules: The lipid injection module is used to automatically inject the biological lipid sample on demand through program control, and inject the lipid sample to be reacted into the mixer module at a set flow rate and volume; A reaction reagent injection module, used for selecting and controlling the injection of photochemical reaction reagents into the mixer module through a program; A mixer module, which is a device module for mixing the lipid sample to be reacted and the photochemical reaction reagent through program control; A photochemical reaction module, used for photochemical reaction, where the biomolecules in the sample undergo photochemical derivatization reaction; Reaction efficiency monitoring module, which is used to monitor the efficiency of photochemical reactions and to provide real-time monitoring and feedback of samples after photochemical reactions; A collection module is a device module for collecting samples that have completed the photochemical reaction; The calculation control module is used to coordinate the functional timing of each module and realize fully automated reaction analysis.
[0007] The present application proposes a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency, which aims to solve the problems of low flux, poor flexibility, low sample processing efficiency, poor reagent compatibility, unstable injection of reaction reagents, and inability to monitor reaction efficiency and side reaction products in real time in the current ultraviolet reaction system in the field of photochemical research. Figure 1 As shown, it includes a calculation control module, a lipid injection module, a reaction reagent injection module, a mixer module, a photochemical reaction module, a reaction efficiency monitoring module and a collection module. The calculation control module can accurately control the operation of each module through the software automation control program to achieve efficient and stable sample processing and reaction process.
[0008] The system aims to achieve efficient, accurate, and cross-contamination-free lipid photochemical reactions through automated control and multi-channel design, providing strong technical support for research in biochemistry, medicinal chemistry and other fields. By designing a multi-channel reaction unit, the system can process multiple lipid samples at the same time, greatly improving the analysis throughput. At the same time, automated control technology is used to achieve precise regulation of parameters such as reagent ratio, reagent flow rate, wavelength, and real-time monitoring and feedback of samples after reaction, thereby ensuring the accuracy and reliability of the analysis results. In addition, the system is also compatible with the analysis of trace samples, so that scarce or difficult-to-obtain lipid samples can also be effectively analyzed.
[0009] Preferably, the lipid injection module comprises an injection pump and an injection channel; the injection pump is used to achieve adjustable sample injection flow rate, ratio and volume, with an adjustable flow rate range of 0.1-1000 μL / min and an adjustable injection volume of 0.05-1000 μL, and the sample is pushed into the mixer as needed.
[0010] Preferably, the injection channel is selected from any one of a single channel or a multi-channel design, and the number of reaction channels of the injection channel is set according to the reaction flux requirement, the multiple channels do not interfere with each other, and there is no loss of samples before and after the reaction.
[0011] Preferably, the reaction reagent injection module includes an injection pump and an injection channel; the injection channel is distributed on demand according to the reaction reagent requirements, and the reaction reagents include but are not limited to acetone, benzophenone, and diacetylpyridine.
[0012] Preferably, the mixer module is used to achieve mixing of the injection channel of the lipid injection module, the incoming sample to be reacted and the reaction reagent of the reaction reagent injection module; the mixer module comprises a mixing valve and a flow rate sensor; according to the sample to be reacted and the required reagent, the channel of the mixing valve is adjusted to open as needed to achieve mixing of the two, and the total flow rate after mixing can be monitored by the flow rate sensor to adjust the reaction time.
[0013] Preferably, the photochemical reaction module comprises a reaction pipeline and a reaction light source to realize a photochemical reaction; the material of the reaction pipeline includes but is not limited to quartz, polytetrafluoroethylene, and polymethyl methacrylate.
[0014] Preferably, in the photochemical reaction module, the light reaction wavelength of the reaction light source is adjusted according to the actual reaction requirements, the actual reaction wavelength is selected from any one of a single wavelength or multiple wavelengths, and the light reaction wavelength range is 100nm-500nm.
[0015] Preferably, the reaction efficiency monitoring module is a miniature mass spectrometer, which includes but is not limited to an ion trap mass spectrometer or a quadrupole mass spectrometer, which detects the abundance of product ions and substrate ions of the post-reaction sample diverted from the photochemical reaction module, obtains the reaction efficiency value and feeds it back to the calculation control module. Preferably, the collection module matches the corresponding collection volume according to the selected channel volume, is compatible with 0.05-1000 μL collection containers, and the collection between multiple channels does not interfere with each other.
[0016] Preferably, after obtaining the value of the monitoring module, the calculation control module compares it with the target reaction efficiency range, thereby outputting the reaction parameters to be readjusted; it can realize information collection and closed-loop control of sample and reagent injection pumps, mixing valves, light sources, temperature sensors, heat dissipation devices, and liquid leakage sensors.
[0017] In a second aspect, the present application also provides a self-regulating lipid photochemical reaction method with controllable reaction efficiency, which is performed using the above self-regulating lipid photochemical reaction system with controllable reaction efficiency; and comprises the following analysis steps: The system self-checks and controls the temperature to be maintained at 15℃-25℃, with no leakage. Start the calculation control module and the reaction efficiency monitoring module; Load the cleaning solvent into the injection module and the reaction reagent module, and increase the flow rate to clean the injection port and channel; Select the corresponding number of sample channels according to the number of samples to be reacted, and select the number of channels of the reaction reagent injection module according to the type of reaction reagent; Loading the lipid sample to be reacted into the injection module, and loading the reaction reagent into the reaction reagent injection module; Set the initial flow rate and volume of lipid sample loading, as well as the adjustment step, select the corresponding photochemical reaction reagent as needed, adjust the position of the mixing valve connection channel port, set the warning value of the reaction reagent loading flow rate, set the warning value of the reaction light source wavelength, and install the collection container as needed; The lipid sample and the reaction reagent are injected into the mixing valve according to the setting, and after mixing, they flow into the reaction module, and a photochemical reaction occurs in the reaction module; The target reaction efficiency range is set, and the samples undergoing photochemical reaction are monitored in real time by the reaction efficiency monitoring module and fed back to the calculation control module. The system adjusts the reaction reagent loading flow rate, reaction reagent ratio, or reaction light wavelength according to the set target reaction efficiency range to achieve the target reaction efficiency; The post-reaction sample confirmed by the reaction efficiency monitoring module switches the sample flow path to the collection module through the switching valve, matches the corresponding collection method / interface according to the selected reaction channel in the collection module, and stores the collected sample in the corresponding container, otherwise the sample flows into the waste liquid container; Set the temperature of the collection module to keep the samples within the appropriate temperature range; The lipid solution after the reaction flows into the collection module and separates the lipid reaction product and the reaction reagent by liquid-liquid extraction; The lipid reaction products are injected into a mass spectrometry system with tandem mass spectrometry analysis function, including but not limited to triple quadrupole, quadrupole-time of flight, etc., and the lipid structure analysis and relative quantification of lipid structural isomers can be achieved through tandem mass spectrometry analysis and diagnostic ion identification.
[0018] In summary, the technical solution of this application has the following effects: The present application proposes a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency, which can solve the problems existing in the current photochemical reaction system in the field of photochemical research, such as the inability to monitor reaction efficiency in real time, low flux, poor flexibility, low sample processing efficiency, poor reagent compatibility, and unstable injection of reaction reagents.
[0019] The self-regulating lipid photochemical reaction system and method with controllable reaction efficiency proposed in this application realizes efficient, accurate and cross-contamination-free lipid photochemical reactions through automated control and multi-channel design, providing strong technical support for research in the fields of biochemistry and medicinal chemistry. The system can process multiple lipid samples at the same time by designing a multi-channel reaction unit, greatly improving the analysis throughput. At the same time, automated control technology is used to achieve precise regulation of parameters such as reagent ratio and wavelength, and real-time monitoring of samples after reaction, thereby ensuring the accuracy and reliability of the analysis results. In addition, the system is also compatible with the analysis of trace samples, so that scarce or difficult-to-obtain lipid samples can also be effectively analyzed. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 Schematic diagram of the self-regulating lipid photochemical reaction system and method with controllable reaction efficiency of the present application.
[0021] Figure 2 Schematic diagram of a single-channel photochemical reaction of a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency in Example 1.
[0022] Figure 3 This is a flow chart for determining the self-regulating lipid photochemical reaction system with controllable reaction efficiency in Example 1.
[0023] Figure 4 This is a flow chart of the analysis process of the self-regulating lipid photochemical reaction system and method with controllable reaction efficiency in Example 2.
[0024] Figure 5 This is the primary mass spectrum of phospholipid PC (34:1) using acetone as a photochemical reaction reagent based on the self-regulating lipid photochemical reaction system and method with controllable reaction efficiency in Example 3.
[0025] Figure 6 This is the primary mass spectrum of phospholipid PC (34:1) using benzophenone as a photochemical reaction reagent based on the self-regulating lipid photochemical reaction system and method with controllable reaction efficiency in Example 4. DETAILED DESCRIPTION
[0026] The present application is further described in detail below in conjunction with examples, comparative examples and performance testing experiments. These examples should not be construed as limiting the scope of protection claimed in the present application.
[0027] Embodiments of the present application are described in detail below, and these embodiments are shown in the accompanying illustrations. Throughout the document and the accompanying drawings, the same or similar marks are used to indicate components with the same or similar characteristics, and components that perform the same or similar functions. The embodiments described by citing these drawings are intended to be illustrative cases to illustrate the concept of the present invention, rather than to limit the scope of application of the present invention. In short, these examples are intended to illustrate but not to limit the scope of the present invention. The self-regulating lipid photochemical reaction system and method with controllable reaction efficiency proposed in accordance with an embodiment of the present invention will be described below with reference to the accompanying drawings. Example
[0028] Example 1 This embodiment provides a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency.
[0029] like Figure 2 As shown, it is a single-channel structure diagram of the self-regulating lipid photochemical reaction system and method with controllable reaction efficiency of this embodiment. The specific process used is as follows: First, the temperature of the injection module is set by the calculation control module, and the lipid sample to be reacted is placed in a suitable sample bottle according to the volume of the lipid sample to be reacted, and marked in the calculation control module, and a suitable photochemical reaction reagent (taking acetone as an example) is selected according to the needs. According to the needs, the flow rate of the lipid sample to be reacted and the photochemical reaction reagent entering the mixer is set in the calculation control module. In the mixer, the lipid sample to be reacted and the photochemical reaction reagent are mixed in the channel after passing through the mixing valve, and the liquid flow rate of the mixed solution of the lipid sample to be reacted and the photochemical reaction reagent flowing into the photochemical reactor is monitored by the built-in flow rate sensor of the mixing module. According to the requirements, the target reaction efficiency range, the warning value of the reaction reagent loading flow rate, and the warning value of the reaction light source wavelength are set in the calculation control module. The mixed solution of the lipid sample to be reacted and the photochemical reaction reagent flowing in undergoes a photochemical derivatization reaction in the photochemical reactor. At the same time, the temperature of the photochemical reactor can be set in the calculation control module to avoid excessive local temperature caused by long-term irradiation of ultraviolet light, which affects the photochemical reaction. Figure 3 As shown, after the photochemical derivatization reaction is completed, the reaction sample flows into the reaction efficiency monitoring module for real-time monitoring, which not only monitors the target reaction efficiency but also monitors the side reaction products; after the reaction efficiency is met, the sample can pass through the collection module to collect the reacted sample, and the temperature of the collection module can be set in the calculation control module to control the temperature within an appropriate range.
[0030] A feature of the present embodiment is that there is no restriction on the photochemical derivatization reagent used and the wavelength of light of the reaction in the present embodiment, and the photochemical reaction reagents that can be used include but are not limited to acetone, benzophenone, diacetylpyridine, etc., and the wavelength range of the photochemical reaction that can be selected is 100nm-500nm. The parameters of each module and the specific reaction conditions can also be controlled by the calculation control module, and real-time monitoring and control can be performed, while the volume of the lipid sample to be reacted is unlimited. It not only realizes real-time monitoring and feedback of the reaction throughout the process, but also greatly improves the working efficiency of the overall photochemical reaction.
[0031] Example 2 This embodiment provides a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency.
[0032] like Figure 4 As shown, it is a multi-channel analysis flow chart of the self-regulating lipid photochemical reaction system and method with controllable reaction efficiency of the present embodiment. The specific process used is as follows: First, the temperature of the injection module is set by the calculation control module, and the lipid sample to be reacted is placed in a suitable sample bottle according to the volume of the lipid sample to be reacted. Three samples can be injected at the same time and marked in the calculation control module. According to the needs, a suitable photochemical reaction reagent is selected, and multiple samples can select the same photochemical reaction reagent, or different photochemical reaction reagents can be selected. According to the needs, the target reaction efficiency range is set in the calculation control module, and the initial value and warning value of the reaction reagent loading flow rate are set. In the mixer, the lipid sample to be reacted and the photochemical reaction reagent are mixed in the channel after passing through the mixing valve, and the liquid flow rate of the mixed solution of the lipid sample to be reacted and the photochemical reaction reagent flowing into the photochemical reactor is monitored by the built-in flow rate sensor of the mixing module. The mixer is automatically cleaned between the two samples to avoid cross contamination caused by different samples and different reaction reagents. Then, the reaction light source wavelength warning value is set in the calculation control module, and different channels can set different light wavelength warning values, or the same light wavelength warning value can be set. The mixed solution of the lipid sample to be reacted and the photochemical reaction reagent flows in the photochemical reactor for photochemical derivatization reaction. At the same time, the temperature of the photochemical reactor can be set in the calculation and control module to avoid long-term irradiation of the ultraviolet lamp causing excessive local temperature and affecting the photochemical reaction. After the photochemical derivatization reaction is completed, the reaction sample flows into the reaction efficiency monitoring module for real-time monitoring, which not only monitors the target reaction efficiency but also monitors the side reaction products; after the reaction efficiency is met, the reaction-completed sample can be collected through the collection module, and then the lipid reaction product and the reaction reagent are separated by liquid-liquid extraction. The lipid reaction product is injected into the mass spectrometry system, and the cascade mass spectrometry analysis can achieve efficient analysis of lipids containing fine structures such as C=C. At the same time, the temperature of the collection module can be set in the calculation and control module to control the temperature within an appropriate range.
[0033] A feature of this embodiment is that multiple channels are used to carry out photochemical reactions, which greatly improves the throughput of lipid sample analysis.
[0034] Example 3 This embodiment provides a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency.
[0035] like Figure 5 The figure shows the primary mass spectrum of the photochemical reaction of lipid PC (34:1) based on Example 1. It is a single-channel photochemical reaction, and the reaction reagent is acetone. The specific steps are as follows: 1. In the control software, set the temperature of the injection module to 15°C, the temperature of the collection module to 15°C, and the temperature of the reactor to 25°C, and wait for the temperature to balance to the set temperature.
[0036] 2. Set the target reaction yield in the control software to 15%-20%.
[0037] 3. Turn on the power of the UV lamp, set the reaction wavelength to 254nm, and preheat and balance the UV lamp for 10 minutes.
[0038] 4. Prepare 100 μL of 1 μg / mL PC (34:1) standard and place it in a 1 mL injection vial for later use.
[0039] 5. Take 100 mL of acetone as a photochemical reaction reagent and set aside.
[0040] 6. In the control software, set the initial flow rate of lipid sample injection to 5 μL / min, the speed adjustment step to 1 μL / min, the injection time to 2 min, the initial flow rate of acetone to 0.1 mL / min, and the injection time to 2 min.
[0041] 7. After the UV lamp balance is completed, click to start sampling.
[0042] 8. The sample obtained by the reaction at the initial flow rate flows into the micro mass spectrometry reaction monitoring module, and the substrate peak and product peak m / z 760.5 and m / z 818.6 are identified, and the reaction yield is calculated to be 13%.
[0043] 9. The reaction yield monitored in the calculation control module is 13% lower than the set range, that is, the sample flow rate is adjusted to 4 μL / min through the control module.
[0044] 10. The sample obtained by the reaction at a sample flow rate of 4 μL / min flowed into the micro mass spectrometry reaction monitoring module, and the substrate peak and product peak m / z 760.5 and m / z 818.6 were identified, and the reaction yield was calculated to be 15.2%.
[0045] 11. The reaction yield of 15.2% monitored in the calculation and control module is within the set range, that is, the liquid path of the post-reaction solution is switched to the collection module.
[0046] 12. According to the above reaction time and solution flow rate, the volume is less than 1mL, and the collection module automatically matches a 1mL collection bottle for collection. Because acetone can directly enter the mass spectrometer, there is no need for liquid-liquid extraction to extract lipids. Then, the product after the PC (34:1) reaction is pushed into the mass spectrometer by direct injection for mass spectrometry analysis.
[0047] Figure 5 Two mass spectrometry peaks can be clearly observed, namely m / z 760.5 and m / z 818.6. The mass spectrometry peak of PC(34:1) in the positive mode is m / z 760.5. With acetone as the photochemical reaction reagent, 58Da of acetone molecular weight will be added to the original mass after the reaction. Therefore, after the photochemical reaction of PC(34:1), the mass of its product is 818.6Da, that is, Figure 4 Another mass spectrum peak m / z 818.6 is shown in the figure. The reason is that after the photochemical reaction of PC(34:1), acetone is added to its structure, so the mass of the reaction product needs to be added with the mass of acetone 58Da.
[0048] Example 4 This embodiment provides a self-regulating lipid photochemical reaction system and method with controllable reaction efficiency.
[0049] like Figure 6 The figure shows the primary mass spectrum of the photochemical reaction of lipid PC (34:1) based on Example 1. It is a single-channel photochemical reaction, and the reaction reagent is benzophenone. The specific steps are as follows: 1. In the control software, set the temperature of the injection module to 15°C, the temperature of the collection module to 15°C, and the temperature of the reactor to 25°C, and wait for the temperature to balance to the set temperature.
[0050] 2. Set the target reaction yield in the control software to 16%-23%.
[0051] 3. Turn on the power of the UV lamp, set the reaction wavelength to 254nm, and preheat and balance the UV lamp for 10 minutes.
[0052] 4. Prepare 100 μL of 1 μg / mL PC (34:1) standard and place it in a 1 mL injection vial for later use.
[0053] 5. Prepare 100 mL of 10 mM benzophenone methanol solution as a photochemical reaction reagent and set aside.
[0054] 6. In the control software, set the lipid sample injection flow rate to 2 μL / min, the injection time to 5 min, the benzophenone flow rate to 0.1 mL / min, and the injection time to 5 min.
[0055] 7. After the UV lamp balance is completed, click to start sampling.
[0056] 8. The sample obtained by the reaction at the initial flow rate flows into the micro mass spectrometry reaction monitoring module, and the substrate peak and product peak m / z 760.5 and m / z 942.5 are identified, and the reaction yield is calculated to be 15.6%.
[0057] 9. The reaction yield of 15.6% monitored in the calculation control module is lower than the set range, that is, the sample flow rate is adjusted to 1.5 μL / min through the control module.
[0058] 10. The sample obtained by the reaction at a sample flow rate of 1.5 μL / min flowed into the micro mass spectrometry reaction monitoring module, and the substrate peak and product peak m / z 760.5 and m / z 942.5 were identified, and the reaction yield was calculated to be 17.4%.
[0059] 11. The reaction yield of 17.4% monitored in the calculation and control module is within the set range, that is, the liquid path of the post-reaction solution is switched to the collection module.
[0060] 12. According to the above reaction time and solution flow rate, the volume is less than 1mL, and the collection module automatically matches a 1mL collection bottle for collection. Because benzophenone can directly enter the mass spectrometer, there is no need for liquid-liquid extraction to extract lipids. Then, the product after the PC (34:1) reaction is pushed into the mass spectrometer by direct injection for mass spectrometry analysis.
[0061] Figure 6 Two mass spectrometry peaks can be clearly observed, namely m / z 760.5 and m / z 942.5. The mass spectrometry peak of PC(34:1) in the positive mode of mass spectrometry is m / z 760.5. With benzophenone as the photochemical reaction reagent, after the reaction, the molecular weight of benzophenone of 182Da will be added to the original mass. Therefore, after the photochemical reaction of PC(34:1), the mass of its product is 942.5Da, that is, Figure 5 Another mass spectrum peak m / z 942.5 is shown in the figure. The reason is that after the photochemical reaction of PC(34:1), benzophenone is added to its structure, so the mass of the reaction product needs to be added with the mass of benzophenone 182Da.
[0062] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.
Claims
1. A self-regulating lipid photochemical reaction system with controllable reaction efficiency, characterized in that: Includes the following modules: The lipid injection module is used to automatically inject the biological lipid sample on demand through program control, and inject the lipid sample to be reacted into the mixer module at a set flow rate and volume; A reaction reagent injection module, used for selecting and controlling the injection of photochemical reaction reagents into the mixer module through a program; A mixer module, which is a device module for mixing the lipid sample to be reacted and the photochemical reaction reagent through program control; A photochemical reaction module, used for photochemical reaction, where the biomolecules in the sample undergo photochemical derivatization reaction; Reaction efficiency monitoring module, which is used to monitor the efficiency of photochemical reactions and to provide real-time monitoring and feedback of samples after photochemical reactions; A collection module is a device module for collecting samples that have completed the photochemical reaction; The calculation control module is used to coordinate the functional timing of each module and realize fully automated reaction analysis.
2. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: The lipid injection module comprises an injection pump and an injection channel; the injection pump is used to achieve adjustable sample injection flow rate and volume, the flow rate range is 0.1-1000 μL / min, the injection volume is 0.05-1000 μL, and the sample is pushed into the mixer as needed; the injection channel is selected from any one of a single channel or a multi-channel design, the injection channel sets the number of reaction channels according to the reaction flux requirements, the multi-channels do not interfere with each other, and there is no loss before and after the sample reaction.
3. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: The reaction reagent injection module includes an injection pump and an injection channel; the injection channel is distributed on demand according to the reaction reagent requirements, and the reaction reagents include but are not limited to acetone, benzophenone, and diacetylpyridine.
4. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: The mixer module is used to achieve mixing of the injection channel of the lipid injection module, the incoming sample to be reacted and the reaction reagent of the reaction reagent injection module; the mixer module comprises a mixing valve and a flow rate sensor; according to the sample to be reacted and the required reagent, the channel of the mixing valve is adjusted to open as required to achieve mixing of the two, and the total flow rate after mixing can be monitored by the flow rate sensor to adjust the reaction time.
5. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: The photochemical reaction module includes a reaction pipeline and a reaction light source to realize a photochemical reaction; the material of the reaction pipeline includes but is not limited to quartz, polytetrafluoroethylene, and polymethyl methacrylate.
6. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: In the photochemical reaction module, the light reaction wavelength of the reaction light source is adjusted according to the actual reaction requirements, and the actual reaction wavelength is selected from any one of a single wavelength or multiple wavelengths, and the light reaction wavelength range is 100nm-500nm.
7. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: The reaction efficiency monitoring module is a miniature mass spectrometer, which includes but is not limited to an ion trap mass spectrometer or a quadrupole mass spectrometer, which detects the abundance of product ions and substrate ions of the post-reaction sample diverted from the photochemical reaction module, obtains the reaction efficiency value and feeds it back to the calculation control module.
8. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: The collection module matches the corresponding collection volume according to the selected channel volume, is compatible with 0.05-1000 μL collection containers, and the collection between multiple channels does not interfere with each other.
9. The self-regulating lipid photochemical reaction system with controllable reaction efficiency according to claim 1, characterized in that: After obtaining the value of the monitoring module, the calculation control module compares it with the target reaction efficiency range, thereby outputting the reaction parameters to be readjusted; it can realize information collection and closed-loop control of sample and reagent injection pumps, mixing valves, light sources, temperature sensors, heat dissipation devices, and liquid leakage sensors.
10. A self-regulating lipid photochemical reaction method with controllable reaction efficiency, characterized in that: The method is carried out using the self-regulating lipid photochemical reaction system with controllable reaction efficiency according to any one of claims 1 to 9; The analysis steps include: The system self-checks and controls the temperature to be maintained at 15℃-25℃, with no leakage. Start the calculation control module and the reaction efficiency monitoring module; Load the cleaning solvent into the injection module and the reaction reagent module, and increase the flow rate to clean the injection port and channel; Select the corresponding number of sample channels according to the number of samples to be reacted, and select the number of channels of the reaction reagent injection module according to the type of reaction reagent; Loading the lipid sample to be reacted into the injection module, and loading the reaction reagent into the reaction reagent injection module; Set the initial flow rate and volume of lipid sample loading, as well as the adjustment step, select the corresponding photochemical reaction reagent as needed, adjust the position of the mixing valve connection channel port, set the warning value of the reaction reagent loading flow rate, set the warning value of the reaction light source wavelength, and install the collection container as needed; The lipid sample and the reaction reagent are injected into the mixing valve according to the setting, and after mixing, they flow into the reaction module, and a photochemical reaction occurs in the reaction module; The target reaction efficiency range is set, and the samples undergoing photochemical reaction are monitored in real time by the reaction efficiency monitoring module and fed back to the calculation control module. The system adjusts the reaction reagent loading flow rate, reaction reagent ratio, or reaction light wavelength according to the set target reaction efficiency range to achieve the target reaction efficiency; The post-reaction sample confirmed by the reaction efficiency monitoring module switches the sample flow path to the collection module through the switching valve, matches the corresponding collection method / interface according to the selected reaction channel in the collection module, and stores the collected sample in the corresponding container, otherwise the sample flows into the waste liquid container; Set the temperature of the collection module to keep the samples within the appropriate temperature range; The lipid solution after the reaction flows into the collection module and separates the lipid reaction product and the reaction reagent by liquid-liquid extraction; The lipid reaction products are injected into a mass spectrometry system with tandem mass spectrometry analysis function, including but not limited to triple quadrupole, quadrupole-time of flight, etc., and lipid structure analysis and relative quantification of lipid structural isomers can be achieved through tandem mass spectrometry analysis and diagnostic ion identification.