CN118275485A - Liquid phase free radical real-time in-situ detection device and detection method thereof - Google Patents

Abstract

The present invention discloses a liquid-phase free radical real-time in-situ detection device and a detection method thereof. The device comprises a free radical detection module, a computer control module, a liquid flow control module, a constant temperature control module, a reaction system holding container and a free radical scavenger holding container, wherein the reaction system holding container and the free radical scavenger holding container are both arranged in the constant temperature control module; the free radical detection module comprises an electron paramagnetic resonance spectrometer, a sample tube, a quartz capillary and a silicone sleeve, wherein the sample tube is arranged in a detection pool of a resonant cavity of the electron paramagnetic resonance spectrometer, the quartz capillary and the silicone sleeve are both located in the sample tube, the silicone sleeve is sleeved on the quartz capillary, one end of the quartz capillary is connected to the reaction system holding container and the free radical scavenger holding container through a pipeline, and the liquid flow control module is arranged on the pipeline. The present application can greatly improve the detection efficiency of free radicals while realizing the in-situ real-time detection of free radicals.

Description

Liquid phase free radical real-time in-situ detection device and detection method thereof

Technical Field

The present application relates to the technical field of analysis and testing, and in particular to a real-time in-situ detection device for liquid-phase free radicals and a detection method thereof.

Background technique

Free radicals refer to atoms or groups with unpaired electrons formed by homolytic cleavage of covalent bonds in molecules of compounds under external conditions such as light and heat, also known as "free radicals". The existence of unpaired electrons brings two characteristics of free radicals. First, unpaired electrons have a strong tendency to pair, so they have high reactivity. Free radicals are very easy to react such as dimerization, hydrogen abstraction, oxidation, disproportionation, etc. Except for very few cases, free radicals are difficult to exist stably; second, there is a spin magnetic moment brought by unpaired electrons, which is also the main basis for the detection of free radicals by electron paramagnetic resonance technology. Free radical reactions play an important role in combustion, atmospheric chemistry, polymerization, plasma chemistry, biochemistry and various other disciplines. Modern biomedical research has also shown that free radicals are closely related to human aging and a variety of diseases including heart disease, Alzheimer's disease, Parkinson's disease and tumors. Therefore, free radical research has always been an important research direction, and the detection of free radicals is a very basic and important work.

At present, in traditional technologies, the methods for detecting free radicals mainly include electron paramagnetic resonance (EPR), high performance liquid chromatography (HPLC), spectrophotometry, fluorescence spectroscopy and electrochemical methods. Among them, HPLC needs to be used in conjunction with other detection technologies to perform accurate quantitative analysis; spectrophotometry, fluorescence spectroscopy and electrochemical methods quantitatively detect free radicals by changes in absorbance, luminescence intensity and electrical signals before and after the reaction. Among these methods, the most commonly used method is electron paramagnetic resonance (EPR). The usual operation steps are to manually sample and add free radical scavengers, then load them into capillaries and put them into the resonant cavity of the electron paramagnetic resonance spectrometer for testing. The above operations are cumbersome and time-consuming, and also bring many problems such as sampling, capturing, and loading samples. The process takes a certain amount of time. For some reactions with very fast rates, due to the short life of the reaction intermediates, manual operation is difficult to achieve real-time in-situ monitoring, and even for some reactions with slow reaction rates and long free radical life, manual operation is difficult to achieve continuous monitoring of the reaction system.

Summary of the invention

Based on this, it is necessary to provide a liquid phase free radical real-time in-situ detection device. The liquid phase free radical real-time in-situ detection device of the present invention can realize the in-situ real-time detection of free radicals while greatly improving the detection efficiency of free radicals.

An embodiment of the present application provides a real-time in-situ detection device for liquid-phase free radicals.

A liquid phase free radical real-time in-situ detection device, comprising a free radical detection module, a computer control module, a liquid flow control module, a constant temperature control module, a reaction system holding container and a free radical scavenger holding container;

Wherein, the reaction system holding container and the free radical scavenger holding container are both arranged in the constant temperature control module;

The free radical detection module comprises an electron paramagnetic resonance spectrometer, a sample tube, a quartz capillary and a silicone sleeve, wherein the sample tube is arranged in a detection pool of a resonant cavity of the electron paramagnetic resonance spectrometer, the quartz capillary and the silicone sleeve are both located in the sample tube, the silicone sleeve is sleeved on the quartz capillary, one end of the quartz capillary is connected to the reaction system holding container and the free radical scavenger holding container through a pipeline, the liquid flow control module is arranged on the pipeline, and the other end of the quartz capillary is used to connect to a waste liquid collection pool;

The free radical detection module, the liquid flow control module and the constant temperature control module are electrically connected to the computer control module respectively.

In some embodiments, the constant temperature control module includes a constant temperature water bath.

In some embodiments, the pipeline is a silicone hose, the reaction system holding container and the free radical scavenger holding container are respectively connected to the quartz capillary through the silicone hose, and the liquid flow control module is arranged on the silicone hose.

In some embodiments, the liquid flow control module is a peristaltic pump, which includes at least two pump heads. The two pump heads of the peristaltic pump can respectively realize the transportation and flow control of the liquid in the reaction system container and the liquid in the free radical scavenger container.

In some embodiments, the real-time in-situ detection device for liquid-phase free radicals also includes a three-way valve, and the pipeline connected to the reaction system container and the pipeline connected to the free radical scavenger container are respectively connected to the three-way valve, and the three-way valve is also connected to the quartz capillary.

In some embodiments, the silicone sleeve is between the sample tube and the quartz capillary and is in contact with and fits with the sample tube and the quartz capillary at the same time.

In some embodiments, the liquid-phase free radical real-time in-situ detection device further includes a waste liquid collection pool, and the other end of the quartz capillary is used to be connected to the waste liquid collection pool.

An embodiment of the present application also provides a real-time in-situ detection method for liquid-phase free radicals.

A method for real-time in-situ detection of liquid-phase free radicals, using the real-time in-situ detection device of liquid-phase free radicals, comprises the following steps:

The sample to be tested is placed in a reaction system container, the free radical scavenging agent is placed in a free radical scavenging agent container, and the temperature of the thermostatic control module is controlled to a predetermined temperature;

The pump speed of the liquid flow control module is set, and the operation of the liquid flow control module is controlled by the computer control module to achieve that the sample to be tested in the reaction system holding container is mixed through the pipeline and then enters the quartz capillary of the free radical detection module, or the sample to be tested in the reaction system holding container and the free radical scavenging agent in the free radical scavenging agent holding container are mixed through the pipeline and then enter the quartz capillary of the free radical detection module;

The computer control module controls the electron paramagnetic resonance spectrometer to detect the liquid in the quartz capillary and collect and generate a spectrum.

In some embodiments, the free radical capture agent includes one of DMPO (5,5-dimethyl-1-pyrroline-N-oxide), MNP (2-methyl-2-nitrosopropane dimer), PBN (N-tert-butyl-α-phenylnitrone), 1-oxy-4-pyridyl-N-tert-butyl nitroxide (POBN), TEMPO (2,2,6,6-tetramethylpiperidinyl oxide), and BMPO (5-tert-butyloxycarbonyl-5-methyl-1-pyrroline-N-oxide).

In some embodiments, the method for real-time in-situ detection of liquid-phase free radicals further comprises the following steps:

When a capture reagent is not needed, that is, for some relatively stable free radicals that can be directly detected, the sample to be tested in the reaction system container is mixed through the pipeline and then enters the quartz capillary of the free radical detection module for detection. After the detection is completed, the quartz capillary is pumped back to the reaction system container through the liquid flow control module;

When a capture reagent is needed, the sample to be tested in the reaction system holding container and the free radical capture reagent in the free radical capture reagent holding container are mixed through a pipeline and then enter the waste liquid collection pool.

The above-mentioned real-time in-situ detection device for liquid-phase free radicals realizes the real-time capture of liquid-phase free radicals by driving the sample liquid to be tested and the free radical capture reagent to flow and mix. The mixed liquid passes through the resonant cavity of the electron paramagnetic resonance spectrometer, and the captured free radicals are detected in-situ by the electron paramagnetic resonance spectrometer. In the present application, the flow rate of the sample to be tested and the free radical capture reagent, and the mixing ratio between the sample to be tested and the free radical capture reagent can be controlled, which greatly improves the detection efficiency of free radicals while realizing the in-situ real-time detection of free radicals.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

In order to more completely understand the present application and its beneficial effects, the following description will be given in conjunction with the accompanying drawings. In the following description, the same reference numerals represent the same parts.

FIG1 is a schematic diagram of a real-time in-situ detection device for liquid-phase free radicals according to an embodiment of the present invention;

FIG. 2 is a partial structural schematic diagram of a device for real-time in-situ detection of liquid-phase free radicals according to an embodiment of the present invention.

Description of Reference Numerals

10. Real-time in-situ detection device for liquid-phase free radicals; 100. Free radical detection module; 101. Electron paramagnetic resonance spectrometer; 1011. Resonant cavity; 102. Sample tube; 103. Quartz capillary; 104. Silicone sleeve; 200. Computer control module; 300. Liquid flow control module; 400. Constant temperature control module; 500. Reaction system container; 600. Free radical scavenger container; 700. Three-way valve; 800. Waste liquid collection pool; 900. Silicone hose.

Detailed ways

In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

In the description of the present invention, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, and "above", "below", "within", etc. are understood to include the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

In the present application, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values in the numerical interval is considered to be continuous, and includes the two numerical endpoints (i.e., the minimum value and the maximum value) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to an integer in the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints, which is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical range disclosed in the present application should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows for a broad range of quantitative intervals such as percentage intervals, ratio intervals, and ratio intervals.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and/or" used herein includes any and all combinations of one or more of the related listed items.

The embodiment of the present application provides a liquid-phase free radical real-time in-situ detection device 10 to solve the problems of manual sampling operation in the prior art when the electron paramagnetic resonance method (EPR) is used to detect free radicals, which is cumbersome and time-consuming, and the problem that for some reactions with very fast rates, it is difficult to achieve real-time in-situ monitoring by manual operation due to the short life of the reaction intermediates. The liquid-phase free radical real-time in-situ detection device 10 will be described below in conjunction with the accompanying drawings.

The real-time in-situ detection device 10 for liquid-phase free radicals provided in the embodiment of the present application is exemplified in Figure 1, which is a schematic diagram of the structure of the real-time in-situ detection device 10 for liquid-phase free radicals provided in the embodiment of the present application. The real-time in-situ detection device 10 for liquid-phase free radicals provided in the present application can be used for real-time in-situ detection of liquid-phase free radicals.

In order to more clearly illustrate the structure of the liquid-phase free radical real-time in-situ detection device 10 , the liquid-phase free radical real-time in-situ detection device 10 will be introduced below with reference to the accompanying drawings.

For example, please refer to FIG. 1 , which is a schematic structural diagram of a liquid-phase free radical real-time in-situ detection device 10 provided in an embodiment of the present application.

A liquid phase free radical real-time in-situ detection device 10 includes a free radical detection module 100, a computer control module 200, a liquid flow control module 300, a constant temperature control module 400, a reaction system container 500 and a free radical scavenger container 600.

Among them, the reaction system holding container 500 and the free radical scavenger holding container 600 are both arranged in the constant temperature control module 400. The free radical detection module 100 includes an electron paramagnetic resonance spectrometer 101, a sample tube 102, a quartz capillary 103 and a silicone sleeve 104. The sample tube 102 is arranged in the detection pool of the resonant cavity 1011 of the electron paramagnetic resonance spectrometer 101. The quartz capillary 103 and the silicone sleeve 104 are both located in the sample tube 102. The silicone sleeve 104 is sleeved on the quartz capillary 103. One end of the quartz capillary 103 is connected to the reaction system holding container 500 and the free radical scavenger holding container 600 through a pipeline. The liquid flow control module 300 is arranged on the pipeline. The other end of the quartz capillary 103 is used to connect to the waste liquid collection pool 800;

The free radical detection module 100 , the liquid flow control module 300 and the constant temperature control module 400 are electrically connected to the computer control module 200 , respectively.

In some embodiments, the sample tube 102 is an EPR quartz sample tube 102 .

In some embodiments, the constant temperature control module 400 includes a constant temperature water bath, and the temperature of the constant temperature water bath can be set according to actual needs.

In some embodiments, the pipeline is a silicone hose 900, the reaction system container 500 and the free radical scavenger container 600 are respectively connected to the quartz capillary 103 through the silicone hose 900, and the liquid flow control module 300 is arranged on the silicone hose 900. The silicone hose 900 can cooperate with the liquid flow control module 300 such as a peristaltic pump, and the liquid flow control module 300 realizes the delivery of liquid through peristalsis.

In some embodiments, the liquid flow control module 300 is a peristaltic pump. The peristaltic pump includes at least two pump heads. The two pump heads of the peristaltic pump can respectively realize the delivery and flow control of the liquid in the reaction system container 500 and the liquid in the free radical scavenger container 600.

In some embodiments, the liquid-phase free radical real-time in-situ detection device 10 further includes a three-way valve 700. The pipeline connected to the reaction system container 500 and the pipeline connected to the free radical scavenger container 600 are respectively connected to the three-way valve 700, and the three-way valve 700 is also connected to the quartz capillary 103.

In some embodiments, as shown in FIG. 2 , the silicone sleeve 104 is between the sample tube 102 and the quartz capillary 103 and is in contact with and fits with the sample tube 102 and the quartz capillary 103 at the same time.

In some embodiments, the liquid phase free radical real-time in-situ detection device 10 further includes a waste liquid collection pool 800. The other end of the quartz capillary 103 is used to be connected to the waste liquid collection pool 800.

An embodiment of the present application also provides a method for real-time in-situ detection of liquid-phase free radicals.

It should be noted that, in this article, unless otherwise specified, each reaction step can be carried out in the order described herein, or can be carried out in a non-sequential manner. For example, other steps may be included between each reaction step, and the order of the reaction steps may also be appropriately swapped. This can be determined by the technician based on conventional knowledge and experience. Preferably, the reaction method herein is carried out sequentially.

A method for real-time in-situ detection of liquid-phase free radicals, using a real-time in-situ detection device 10 for liquid-phase free radicals, comprises the following steps:

S1. Place the sample to be tested in the reaction system container 500, place the free radical scavenging agent in the free radical scavenging agent container 600, and control the temperature of the constant temperature control module 400 to a predetermined temperature.

S2. Set the pump speed of the liquid flow control module 300, and control the operation of the liquid flow control module 300 through the computer control module 200 to achieve that the sample to be tested in the reaction system holding container 500 is mixed through the pipeline and then enters the quartz capillary 103 of the free radical detection module 100, or the sample to be tested in the reaction system holding container 500 and the free radical scavenging agent in the free radical scavenging agent holding container 600 are mixed through the pipeline and then enter the quartz capillary 103 of the free radical detection module 100.

S3. The computer control module 200 controls the electron paramagnetic resonance spectrometer 101 to detect the liquid in the quartz capillary 103 and collect and generate a spectrum.

In some of the embodiments, the free radical scavenging agent includes one of DMPO (5,5-dimethyl-1-pyrroline-N-oxide), MNP (2-methyl-2-nitrosopropane dimer), PBN (N-tert-butyl-α-phenylnitrone), 1-oxy-4-pyridyl-N-tert-butylnitroxide (POBN), TEMPO (2,2,6,6-tetramethylpiperidinyloxide), and BMPO (5-tert-butyloxycarbonyl-5-methyl-1-pyrroline-N-oxide).

In some embodiments, the method for real-time in-situ detection of liquid-phase free radicals further comprises the following steps:

When a capture reagent is not needed, that is, for some relatively stable free radicals that can be directly detected, the sample to be tested in the reaction system container 500 is mixed through the pipeline and then enters the quartz capillary 103 of the free radical detection module 100 for detection. After the detection is completed, the quartz capillary 103 is pumped back to the reaction system container 500 through the liquid flow control module 300;

When a capture reagent is needed, the sample to be tested in the reaction system container 500 and the free radical capture reagent in the free radical capture reagent container 600 are mixed through a pipeline and then enter the waste liquid collection tank 800.

Example 1

The present embodiment provides a liquid-phase free radical real-time in-situ detection device 10, comprising a free radical detection module 100, a computer control module 200, a liquid flow control module 300, a constant temperature control module 400, a reaction system container 500, a free radical scavenger container 600, a three-way valve 700, and a waste liquid collection pool 800. The constant temperature control module 400 is a constant temperature water bath. The liquid flow control module 300 is a peristaltic pump.

The reaction system container 500 and the free radical scavenger container 600 are both arranged in the constant temperature control module 400. The free radical detection module 100 includes an electron paramagnetic resonance spectrometer 101, a sample tube 102, a quartz capillary 103 and a silicone sleeve 104. The sample tube 102 is an EPR quartz sample tube 102. The sample tube 102 is arranged in the detection pool of the resonant cavity 1011 of the electron paramagnetic resonance spectrometer 101. The quartz capillary 103 and the silicone sleeve 104 are both located in the sample tube 102. The silicone sleeve 104 is sleeved on the quartz capillary 103.

The reaction system holding container 500 is connected to an interface of the three-way valve 700 through a silicone hose 900, the free radical capture agent holding container 600 is connected to another joint of the three-way valve 700 through a silicone hose 900, and the third joint of the three-way valve 700 is connected to the quartz capillary 103. The peristaltic pump includes two pump heads. The two pump heads of the peristaltic pump can respectively realize the transportation and flow control of the liquid in the reaction system holding container 500 and the liquid in the free radical capture agent holding container 600. The other end of the quartz capillary 103 is connected to the waste liquid collection tank 800. The free radical detection module 100, the liquid flow control module 300 and the constant temperature control module 400 are electrically connected to the computer control module 200 respectively.

Example 2

This embodiment provides a real-time in-situ detection method for liquid-phase free radicals.

This example is described by taking the detection of free radicals in the reaction process of laccase and hydroquinone as an example. Laccase can catalyze the oxidation of hydroquinone to form semiquinone free radicals, which are relatively stable free radicals and can be detected without a free radical scavenger.

S1. Place laccase and hydroquinone in a reaction system container 500 and mix them.

S2. Set the pump speed of the liquid flow control module 300, and control the operation of the liquid flow control module 300 through the computer control module 200 to enable the sample to be tested in the reaction system container 500 to enter the quartz capillary 103 of the free radical detection module 100 through the silicone hose 900.

S3, adjusting the relevant parameters of the electron paramagnetic resonance spectrometer 101, the semiquinone free radicals are detected in real time in the resonant cavity 1011, and the spectrum is collected continuously for a long time, that is, the trend of the change of the semiquinone free radical concentration over time can be seen, and the study of the enzyme catalytic reaction kinetics can be realized. The computer control module 200 controls the electron paramagnetic resonance spectrometer 101 to detect the liquid in the quartz capillary 103 and collect and generate the spectrum.

S4. After passing through the detection pool, the sample to be tested can flow back into the reaction system container 500, realizing the cyclic real-time detection of a small amount of sample; further, by adjusting the temperature of the constant temperature water bath of the constant temperature control module 400, the concentration of semiquinone free radicals under different temperature conditions in the same time can be measured, and the effect of temperature on laccase activity can also be studied.

Example 3

This embodiment provides a real-time in-situ detection method for liquid-phase free radicals.

This embodiment is described by taking the detection of hydroxyl radicals generated during the reaction of hydrogen peroxide and divalent iron ion solution as an example. Hydrogen peroxide and divalent iron ion solution can react to produce hydroxyl radicals, namely Fenton reaction, in which hydroxyl radicals are very active free radicals, and direct electron paramagnetic detection cannot detect signals, and can only be detected after being captured by a free radical scavenger.

S1. Mix the dilute hydrogen peroxide solution and the divalent iron ion solution reaction system in a container 500. Place a free radical scavenger in a free radical scavenger container 600.

S2. Set the pump speed of the liquid flow control module 300, and control the operation of the liquid flow control module 300 through the computer control module 200 to achieve the goal that the sample to be tested in the reaction system container 500 and the free radical scavenger in the free radical scavenger container 600 are mixed at the three-way valve 700 through the silicone hose 900 and then enter the quartz capillary 103 of the free radical detection module 100.

S3, adjusting the relevant parameters of the electron paramagnetic resonance spectrometer 101, the hydroxyl radicals captured by the free radical scavenger are detected in real time in the resonant cavity 1011, and the spectrum is collected continuously for a long time, that is, the trend of the hydroxyl radical concentration changing with time can be seen, and the study of the enzyme catalytic reaction kinetics is realized. The computer control module 200 controls the electron paramagnetic resonance spectrometer 101 to detect the liquid in the quartz capillary 103 and collect and generate the spectrum.

S4. After passing through the detection pool, the sample to be tested enters the waste liquid collection pool 800; further, the temperature of the constant temperature water bath of the constant temperature control module 400 is adjusted to measure the concentration of hydroxyl radicals under different temperature conditions in the same time, and the influence of temperature on the Fenton reaction can also be studied.

In summary, the above-mentioned liquid-phase free radical real-time in-situ detection device 10 realizes the real-time capture of liquid-phase free radicals by driving the sample liquid to be tested and the free radical capture reagent to flow and mix. The mixed liquid passes through the resonant cavity 1011 of the electron paramagnetic resonance spectrometer 101, and the captured free radicals are detected in-situ by the electron paramagnetic resonance spectrometer 101. In the present application, the flow rate of the sample to be tested and the free radical capture reagent, and the mixing ratio between the sample to be tested and the free radical capture reagent can be controlled. While realizing the in-situ real-time detection of free radicals, the detection efficiency of free radicals is greatly improved.

In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims (10)

1. The liquid phase free radical real-time in-situ detection device is characterized by comprising a free radical detection module, a computer control module, a liquid flow control module, a constant temperature control module, a reaction system container and a free radical capturing agent container; the reaction system containing container and the free radical scavenger containing container are arranged in the constant temperature control module; the free radical detection module comprises an electron paramagnetic resonance spectrometer, a sample tube, a quartz capillary tube and a silica gel sleeve, wherein the sample tube is arranged in a detection pool of a resonant cavity of the electron paramagnetic resonance spectrometer, the quartz capillary tube and the silica gel sleeve are both positioned in the sample tube, the silica gel sleeve is sleeved on the quartz capillary tube, one end of the quartz capillary tube is connected with the reaction system containing container and the free radical capturing agent containing container through pipelines, the liquid flow control module is arranged on the pipelines, and the other end of the quartz capillary tube is used for communicating a waste liquid collecting pool; the free radical detection module, the liquid flow control module and the constant temperature control module are respectively and electrically connected with the computer control module.

2. The liquid phase free radical real time in situ detection apparatus according to claim 1, wherein the constant temperature control module comprises a constant temperature water bath.

3. The liquid-phase free radical real-time in-situ detection device according to claim 1, wherein the pipeline is a silica gel hose, the reaction system container and the free radical scavenger container are respectively connected to the silica gel capillary through the silica gel hose, and the liquid flow control module is arranged on the silica gel hose.

4. The liquid-phase free radical real-time in-situ detection device according to any one of claims 1-3, wherein the liquid flow control module is a peristaltic pump, the peristaltic pump comprises at least two pump heads, and the two pump heads of the peristaltic pump can respectively realize the transportation and flow control of the liquid in the reaction system container and the liquid in the free radical capturing agent container.

5. The liquid-phase free radical real-time in-situ detection device according to any one of claims 1 to 3, further comprising a three-way valve, wherein the pipeline connected with the reaction system container and the pipeline connected with the free radical scavenger container are respectively connected with the three-way valve, and the three-way valve is further connected with the quartz capillary tube.

6. The liquid-phase free radical real-time in-situ detection device according to any one of claims 1-3, wherein the silica gel sleeve is between the sample tube and the quartz capillary tube and is in contact fit with the sample tube and the quartz capillary tube at the same time.

7. The liquid-phase free radical real-time in-situ detection device according to any one of claims 1-3, further comprising a waste liquid collecting tank, wherein the other end of the quartz capillary tube is communicated with the waste liquid collecting tank.

8. A liquid phase free radical real-time in-situ detection method, which is characterized by using the liquid phase free radical real-time in-situ detection device according to any one of claims 1-7, comprising the following steps:

Placing a sample to be tested in a reaction system container, placing a free radical capturing agent in the free radical capturing agent container, and controlling the temperature of the constant temperature control module to a preset temperature;

Setting the pump speed of a liquid flow control module, and controlling the action of the liquid flow control module through a computer control module to realize that a sample to be detected in a reaction system container enters a quartz capillary tube of a free radical detection module after being mixed through a pipeline, or that the sample to be detected in the reaction system container and a free radical capture agent in a free radical capture agent container enter the quartz capillary tube of the free radical detection module after being mixed through a pipeline;

And controlling an electron paramagnetic resonance spectrometer to detect the liquid in the quartz capillary through a computer control module and acquiring and generating a spectrogram.

9. The method of real-time in-situ detection of liquid phase free radicals according to claim 8, wherein the free radical capture reagent comprises one of 5, 5-dimethyl-1-pyrroline-N-oxide, 2-methyl-2-nitrosopropane dimer, N-t-butyl- α -phenylnitrone, 1-oxo-4-pyridinyl-N-t-butyloxynitride, 2, 6-tetramethylpiperidine oxide, 5-t-butoxycarbonyl-5-methyl-1-pyrroline-N-oxide.

10. The method for real-time in-situ detection of liquid phase free radicals according to claim 8, further comprising the steps of:

when the reagent is not required to be captured, namely, for some more stable free radicals which can be directly detected, a sample to be detected in the reaction system accommodating container is mixed through a pipeline and then enters a quartz capillary tube of a free radical detection module for detection, and after the detection is finished, the quartz capillary tube is pumped back to the reaction system accommodating container through the liquid flow control module;

When the reagent is required to be captured, the sample to be detected in the reaction system containing container and the free radical capturing reagent in the free radical capturing agent containing container are mixed through a pipeline and then enter the waste liquid collecting tank.