ROS high-efficiency synchronous recognition system combined with multiple molecular probes and application of ROS high-efficiency synchronous recognition system

Abstract

The invention discloses an ROS (reactive oxygen species) efficient synchronous recognition system combined with multiple molecular probes and application. The system comprises a table type vertical rotating device, a first reaction device and a detection device, the first reaction device is detachably fixed on the table type vertical rotating device in a dark place and comprises a reaction medium and a pollutant-probe mixed solution; the reaction medium is periodically kept in a wetting-exposure alternating state in the vertical rotation process; the first reaction device comprises three parallel and independent pollutant-probe mixed solutions which are respectively used for identifying. OH, O2. <-> and < 1 > O2; and the detection device is used for quantitatively determining the residual concentration of a stable product or a probe reagent after the reaction of each probe and the free radical. The method has the advantages of being high in scene adaptability, easy and convenient to operate, low in cost, high in sensitivity, high in matrix interference resistance and the like, and can be widely applied to exploration and research of natural attenuation or enhanced attenuation mechanisms and the like of pollutants in saturated or non-(variable) saturated scenes.

Description

Technical Field

[0001] This invention belongs to the field of environmental monitoring and analysis technology, and specifically relates to a high-efficiency synchronous ROS identification system and its application that combines multiple molecular probes. Background Technology

[0002] Reactive oxygen species (ROS) are a class of highly reactive oxidizing chemical species, mainly including hydroxyl radicals (·OH) and superoxide radicals (O2··OH). - Singlet oxygen () 1 Oxygen spores (O2) and other reactive oxygen species are widely present in natural environmental processes, advanced oxidation water treatment technologies, and biological metabolic systems. In the field of environmental remediation, the amount, reactivity, and lifespan of reactive oxygen species directly determine the degradation efficiency of pollutants. Accurate quantitative detection of these free radicals is a core element in evaluating the effectiveness of remediation technologies and optimizing reaction conditions.

[0003] Traditional ROS detection methods primarily target single free radicals and are susceptible to matrix interference and insufficient sensitivity. Currently, ROS detection methods mainly include electron spin resonance (ESR), chemiluminescence, fluorescent probe methods, and colorimetric methods. Among these, while ESR can directly capture free radical signals, the equipment is expensive, the operation is complex, and the detection cost is high, making it difficult to meet the needs of routine experiments and on-site monitoring. Chemiluminescence has high sensitivity but is easily interfered with by other substances in the system and has poor stability. Although fluorescent probe methods have strong selectivity, some fluorescent probes exhibit photobleaching and have stringent requirements for environmental conditions such as pH and temperature.

[0004] Probe methods, as a quantitative detection technique based on specific chemical reactions, convert free radicals into stable, detectable products by selecting probe molecules with high specificity to the target free radical, and then achieving quantification using chromatographic and spectroscopic analytical methods. This method has advantages such as ease of operation, controllable cost, and good selectivity, and has received widespread attention in the detection of reactive oxygen species (ROS). However, existing probe methods have the following shortcomings: firstly, they lack targeting capabilities for various ROS (·OH, O2·)... - , 1The current integrated detection scheme for O2 requires separate experimental setups and procedures for different free radicals, resulting in low detection efficiency. Secondly, the reaction conditions between probe molecules and free radicals are not standardized, leading to inconsistent product separation and detection parameters, resulting in poor accuracy and repeatability of the detection results. Thirdly, existing detection methods are mostly applicable to static laboratory systems, making it difficult to meet the real-time monitoring needs of free radicals during dynamic reactions. Furthermore, current methods do not consider efficient and accurate ROS identification in actual hydrological environments such as saturated and unsaturated (variable) saturated environments caused by water level fluctuations. Therefore, it is necessary to develop a low-cost, easy-to-operate method that can simulate hydrological changes and simultaneously quantify ·OH and O2· - , 1 The probe method for O2 is of great significance for revealing the synergistic mechanism of free radicals in complex pollution systems. Summary of the Invention

[0005] The purpose of this invention is to provide a system for the simultaneous identification of reactive oxygen species (ROS) based on multiple probes under simulated hydrological changes (saturated and / or (non-)variant saturation) scenarios, and its application, to achieve the identification of hydroxyl radicals (·OH) and superoxide radicals (O2·) under different saturation environments. - Singlet oxygen () 1 The accurate and efficient quantitative simultaneous detection of O2 solves the problems of poor adaptability, cumbersome operation, poor specificity and low detection efficiency of existing detection methods.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows: A high-efficiency synchronous ROS identification system combining multiple molecular probes includes: a benchtop vertical rotating device, a first reaction device, and a detection device; the first reaction device is light-proof and detachably fixed to the benchtop vertical rotating device, and contains a reaction medium and a contaminant-probe mixed solution; the reaction medium of the first reaction device is periodically kept in a wetting-exposure alternating state during vertical rotation to simulate a variable saturation scenario caused by water level fluctuations; the first reaction device contains three parallel and independent contaminant-probe mixed solutions, respectively for ·OH, O2· - , 1 O2 identification; the detection device is used to quantitatively determine the stable product or residual concentration of the probe reagent after the reaction of each probe with a free radical. The vertical rotation is a rotation in the XZ plane, with its rotation axis parallel to the Y-axis. The Y-axis is parallel to the horizontal plane, and the XZ plane is a plane perpendicular to the horizontal plane.

[0007] According to the aforementioned high-efficiency synchronous ROS identification system combining multiple molecular probes, preferably, the system further includes a second reaction device; the first reaction device is light-proof, detachably fixed to a tabletop vertical rotating device, and contains a reaction medium and a contaminant-probe mixed solution; the reaction medium of the second reaction device remains fully submerged during vertical rotation to simulate a saturation scenario; the first and second reaction devices each independently contain three parallel and independent contaminant-probe mixed solutions, respectively for ·OH, O2· - , 1 O2 identification. A tabletop vertical rotating device achieves simultaneous generation and identification of ROS under saturated and unsaturated environments by synchronously fixing n≥1 sets of reaction devices.

[0008] Preferably, the first reaction apparatus and / or the second reaction apparatus further include glass beads, which are used to prevent the reaction medium from being lost with the water flow.

[0009] Preferably, in the aforementioned high-efficiency simultaneous recognition system for ROS using multiple molecular probes, the probe for ·OH recognition is a benzoic acid (BA) probe, and the product p-hydroxybenzoic acid (p-HBA) is quantitatively determined at 254 nm using high-performance liquid chromatography-ultraviolet detection; the probe for O2· - The identified probe was nitroblue tetrazolium (NBT), and residual NBT was quantitatively determined by UV-Vis spectrophotometry at 260 nm; used for identification. 1 The O2 probe was furfuryl alcohol (FFA) probe, and residual FFA was quantitatively determined by high performance liquid chromatography-ultraviolet detector at 230 nm.

[0010] Preferably, the concentration of the benzoic acid (BA) probe in the corresponding contaminant-probe mixed solution is 200 mg·L⁻¹. - ¹; The concentration of the nitroblue tetrazolium (NBT) probe in the corresponding contaminant-probe mixture was 0.1 mmol·L⁻¹. - ¹; The concentration of furfuryl alcohol (FFA) probe in the corresponding contaminant-probe mixed solution was 8.5 mmol·L⁻¹. - ¹.

[0011] According to a specific embodiment of the present invention, the reaction device is a reaction tube containing a reaction medium and a contaminant-probe mixed solution, and the wall of the reaction tube is covered with tin foil to block light.

[0012] Preferably, the system further includes a data processing module and an internal displacement calculation formula to automatically calculate the cumulative yield or generation rate of the target free radical based on the residual concentration of the stable product or probe reagent.

[0013] According to a specific embodiment of the present invention, the reaction medium is an iron-sulfur mineral-quartz sand composite; the pollutants in the pollutant-probe mixed solution are PFOA and GenX, and microorganisms may be selectively added or not added.

[0014] The present invention also provides a method for quantitatively detecting ROS using the system described above, the steps of which include: (1) At room temperature, the reaction apparatus is fixed on a tabletop vertical rotating device and rotated vertically. During the rotation, different probes identify ·OH and O2· respectively. - , 1 O2; (2) After the reaction is complete, collect the liquid sample and use the detection device to quantitatively determine the stable product or residual concentration of the probe reagent after the reaction of each probe with the free radical; (3) Calculate the cumulative yield or generation rate of the target free radical based on the residual concentration of the stable product or probe reagent.

[0015] According to the specific implementation method, in step (1), the table-type vertical rotation device rotates at 20 r·min -1 Rotation speed: 24 h-48 h.

[0016] The system of this invention is applicable to the simultaneous and accurate determination of various reactive oxygen free radicals in scenarios such as natural or enhanced attenuation monitoring, remediation technology research and development, and environmental chemistry under different hydrological fluctuation environments such as saturated and unsaturated environments.

[0017] This invention also provides applications of the system in in-situ ROS identification scenarios, such as natural or enhanced degradation monitoring and the development of advanced oxidation and other environmental remediation technologies. Application scenarios of the identification system include, but are not limited to: advanced oxidation technologies for water remediation, natural / enhanced ROS detection, ROS detection in soil remediation, and other environmental fields (such as...). Figure 5 (As shown). Specific application scenarios include, but are not limited to: monitoring and optimizing ROS production in advanced oxidation water treatment technologies, assessing the contribution of ROS effectiveness in the natural and enhanced degradation remediation processes of soil / groundwater, and studying the degradation mechanisms of ROS in environmental pollutants.

[0018] Specifically, the applications described can be in the detection of naturally / enhanced attenuated ROS, the detection and / or optimization of advanced oxidizing ROS in water remediation, the detection of ROS in soil remediation, and other environmental ROS detection applications. The high-efficiency ROS identification system developed in this invention has the following advantages: (1) The simulated saturated and / or non-(variable) saturated system constructed by the present invention can realize the synchronous identification and monitoring assessment of ROS under actual hydrological environment changes, and has a good match with the actual scenario, which is in line with the actual situation; (2) The highly efficient reactive oxygen species recognition system constructed in this invention can recognize ·OH and O2· - , 1 The independent detection of three reactive oxygen species (O2) simplifies the operation process and improves detection efficiency. (3) By optimizing probe concentration, reaction conditions and detection parameters, the specificity and accuracy of detection were improved, and the influence of interfering substances was effectively reduced; (4) The detection method is based on the mature probe reaction principle, which is simple to operate and cost-controllable, and is suitable for routine laboratory testing and on-site emergency monitoring. (5) According to the quantitative calculation formula in the data processing module, the concentration data of the three reactive oxygen free radicals can be directly converted, reducing intermediate errors and thus improving the repeatability and reliability of the detection results.

[0019] (6) The system of the present invention has the advantages of strong scene adaptability, simple operation, low cost, high sensitivity and strong resistance to matrix interference. It can be widely used in the exploration and research of environmental remediation technology (such as ROS optimization of advanced oxidation process), natural decay or enhanced decay mechanism of pollutants in saturated / or unsaturated (variable) scenarios in soil and groundwater. Attached Figure Description

[0020] Figure 1 The ROS reaction principle corresponding to the three probe detection methods; Figure 2 ROS synchronous recognition simulation system under saturated and non-(variable) saturated scenarios; Figure 3 The identification system for Fe x S y Identification effect of three ROS in mineral matrix; Figure 4 The identification system identifies "microorganism-Fe" x S y The recognition effect of three ROS in the "mineral" interactive matrix; Figure 5 A schematic diagram of the application scenarios of the recognition system. Detailed Implementation

[0021] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings. The following examples will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.

[0022] Example 1: Identification and quantification of iron-sulfur minerals (Fe) in saturated and unsaturated scenarios x Sy ROS generated in the system In saturated scenes, Fe x S y Mineral preparations were completed in advance; in addition, this study constructed a saturation simulation system and simultaneously constructed three sets of probes.

[0023] Construction of the saturation identification system: bottom-up, 14.0 g iron-sulfur mineral-quartz sand composite medium (Fe x S y &QS-0.04 M, where Fe x S y -0.04 M refers to the synthesis of Fe using FeSO4·7H2O and a sulfur source. x S y Minerals, Fe after synthesis x S y The mineral (containing 0.04 mol of Fe) and 8.0 g of glass beads were separated and placed in a 15 mL reaction tube. Figure 2 (a) After adding 7.0 mL of the contaminant-probe mixture, the tube opening was sealed, and the tube was fixed in the rotating device to maintain complete immersion. The contaminants were PFOA and GenX, with a concentration of 0.1 mg·L⁻¹ for both PFOA and GenX. -1 .like Figure 2 As shown, the reaction tube was placed on a benchtop rotator at 20 r·min -1 The reaction tube was rotated at a certain speed for 24 or 48 hours; the reaction tube was covered with tin foil to protect it from light during the reaction; after the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0024] Construction of the saturation control system: Compared with the saturation identification system, 14.0 g of pure quartz sand (QS) was used instead of the iron-sulfur mineral-quartz sand composite medium. The reaction tubes were placed in a benchtop rotator at 20 r·min. -1 The reaction tube was rotated at a certain speed for 24 or 48 hours. During the reaction, the reaction tube wall was covered with tin foil to protect it from light. After the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0025] In unsaturated scenarios, Fe x S y Mineral preparations were completed in advance; in addition, this study constructed an unsaturated simulation system and simultaneously constructed three sets of probes.

[0026] Construction of the unsaturated identification system: From bottom to top, 28.0 g of iron-sulfur mineral-quartz sand composite (Fe x S y&QS-0.02 M, where Fe x S y -0.02 M refers to the synthesis of Fe using FeSO4·7H2O and a sulfur source. x S y Minerals, Fe after synthesis x S y The mineral (containing 0.02 mol of Fe) and 15.0 g of glass beads were placed in a 30 mL reaction tube. Figure 2 In step b), 7.0 mL of the contaminant-probe mixture was added and the tube was sealed, causing the medium to periodically alternate between wetting and exposure during rotation to simulate the variable saturation conditions caused by water level fluctuations. The contaminants were PFOA and GenX, with each PFOA and GenX having a concentration of 0.1 mg·L⁻¹. -1 .like Figure 2 As shown, the reaction tube is placed on a benchtop vertical rotating device at 20 r·min -1 The reaction tube was incubated with shaking for 24 or 48 hours (oxygen was added every 12 hours during the reaction, and then the tube was resealed); the reaction tube was covered with tin foil to protect it from light during the reaction; after the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0027] Construction of the unsaturated control system: Compared with the unsaturated identification system, 28.0 g of pure quartz sand (QS) was used to replace Fe. x S y Mineral-quartz sand composite medium. The reaction tube is placed on a benchtop vertical rotating device at 20 r·min. -1 The reaction mixture was incubated with shaking at a constant speed for 24 or 48 hours. During the reaction, the reaction tube walls were covered with tin foil to protect it from light. After the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0028] Example 2: Identification and quantification of "microorganism-iron-sulfur minerals (Fe)" in saturated and unsaturated scenarios x S y ROS generated in the ")" interaction system In saturated scenes, Fe x S y The mineral-microbial interaction medium was prepared in advance; in addition, this study constructed a saturation simulation system and simultaneously constructed three sets of probes.

[0029] Construction of the saturation identification system: 14.0 g of iron-sulfur mineral-quartz sand composite medium (Fe x S y &QS-0.04 M, where Fex S y -0.04 M refers to the synthesis of Fe using FeSO4·7H2O and a sulfur source. x S y Minerals, Fe after synthesis x S y The mineral (containing 0.04 mol of Fe) and 8.0 g of glass beads were separated and placed in a 15 mL reaction tube. Figure 2 (a) Add 7.0 mL of a contaminant-probe-microorganism mixture (the microorganisms are strains capable of degrading PFOA and GenX, with a concentration of 2.2 g / L). The contaminants are PFOA and GenX, with each at a concentration of 0.1 mg·L⁻¹. -1 The reaction tube was placed on a benchtop rotary table at 20 r·min. -1 The reaction tube was rotated at a certain speed for 24 hours. During the reaction, the reaction tube wall was covered with tin foil to protect it from light. After the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0030] Construction of the saturation control system: Compared with the saturation identification system, 14.0 g of pure quartz sand (QS) was used instead of the iron-sulfur mineral-quartz sand composite medium. The reaction tubes were placed in a benchtop rotator at 20 r·min. -1 The reaction tube was rotated at a certain speed for 24 hours. During the reaction, the reaction tube wall was covered with tin foil to protect it from light. After the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0031] In unsaturated scenarios, Fe x S y The mineral-microbial interaction medium was prepared in advance; in addition, this study constructed an unsaturated simulation system and simultaneously constructed three sets of probes.

[0032] Construction of the unsaturated identification system: 28.0 g of iron-sulfur mineral-quartz sand composite (Fe x S y &QS-0.02 M, where Fe x S y -0.02 M refers to the synthesis of Fe using FeSO4·7H2O and a sulfur source. x S y Minerals, Fe after synthesis x S y The mineral (containing 0.02 mol of Fe) and 15.0 g of glass beads were placed in a 30 mL reaction tube. Figure 2(b) Similarly, add 7.0 mL of a contaminant-probe-microorganism mixture (the microorganisms are strains capable of degrading PFOA and GenX, with a concentration of 2.2 g / L). The contaminants are PFOA and GenX, with each contaminant at a concentration of 0.1 mg·L⁻¹. -1 The reaction tube was placed on a benchtop vertical rotating device at 20 r·min. -1 The reaction was incubated with shaking at a constant speed for 24 hours. During the reaction, the reaction tube walls were covered with tin foil to protect it from light. After the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0033] Construction of the unsaturated control system: Compared with the unsaturated identification system, 28.0 g of pure quartz sand (QS) was used to replace Fe. x S y Mineral-quartz sand composite medium. The reaction tube is placed on a benchtop vertical rotating device at 20 r·min. -1 The reaction was incubated with shaking at a constant speed for 24 hours. During the reaction, the reaction tube walls were covered with tin foil to protect it from light. After the experiment, liquid samples were collected for the detection of ·OH and O2·. - , 1 The amount of O2 generated.

[0034] The quantitative analysis methods for the three ROS in Examples 1 and 2 are as follows (see details of the reaction principle). Figure 1 Quantitative analysis was completed within 90 min, with a detection limit ≤ 0.1 μmol·L⁻¹. - ¹, relative standard deviation < 5%) (1) Quantitative analysis of hydroxyl radicals (·OH) Quantitative analysis of hydroxyl radicals (·OH) was performed using the benzoic acid (BA) probe method. In the presence of ·OH, benzoic acid can be converted into three hydroxybenzoic acid isomers, accounting for 90±5% of the total reaction products. The formation ratio of o-hydroxybenzoic acid (o-HBA), m-hydroxybenzoic acid (m-HBA), and p-hydroxybenzoic acid (p-HBA) was 1.7:2.3:1.2. The cumulative amount of ·OH formed could be estimated by measuring the concentration of p-hydroxybenzoic acid (p-HBA) in the aqueous phase. High-performance liquid chromatography (HPLC) was used to determine the p-hydroxybenzoic acid in the collected aqueous solution samples after the reaction. The instrument parameters were as follows: UV detector, detection wavelength 254 nm; mobile phase: 1% acetic acid aqueous solution and methanol, volume ratio 40:60; flow rate 1.0 mL·min. -1 The chromatographic column was a SunFire-C18 column (3.9×5 mm); the column temperature was 30 ℃; and the injection volume was 50 μL.

[0035] The data processing module, employing a built-in quantitative calculation formula, automatically calculates the cumulative yield of ·OH based on the detected p-HBA product concentration. The calculation formula is as follows: Cumulative OH production = [p-HBA] × 5.87 In this formula, [p-HBA] represents the detected concentration of p-hydroxybenzoic acid (μmol·L⁻¹). -1 ).

[0036] (2) Superoxide anion radical (O2· - Quantitative analysis of ) Nitrogen blue tetrazolium (NBT) colorimetric method was used to detect superoxide anion radicals (O2· - Quantitative analysis was performed. O2· - The yellow nitroblue tetrazolium can be reduced to blue formazan. The collected aqueous solution sample after the reaction was exposed to sunlight for 1 h, then filtered through a 0.22 μm polyvinylidene fluoride (PVDF) membrane to remove precipitates. The solution was diluted 2-fold and the absorbance was measured using a UV-Vis spectrophotometer. The decrease in NBT concentration at 260 nm was used as an indicator to analyze O2· - The percentage of the generated equivalent.

[0037] The data processing module, which employs a built-in quantitative calculation formula, automatically converts the detected remaining NBT concentration into O2· - The generation amount is calculated using the following formula: O2· - Percentage of equivalent generated NBT = (Initial NBT concentration - Remaining NBT concentration) × 2 / Initial NBT concentration In this formula, the NBT concentration is expressed in μmol·L⁻¹. -1 Coefficient 2 is the correction value for the sample dilution factor.

[0038] (3) Singlet oxygen ( 1 Quantitative analysis of O2) Furfuryl alcohol (FFA, concentration 8.5 mmol·L⁻¹) was used. -1 The indicator method for singlet oxygen ( 1 Quantitative analysis of O2 was performed. High-performance liquid chromatography (HPLC) was used to quantitatively detect residual FFA in the collected aqueous solution samples after the reaction. A SunFire-C18 column was used, with a UV detection wavelength of 230 nm. The mobile phase was 85:15 (V:V) deionized water and methanol, and the flow rate was 1.0 mL / min. - ¹, column temperature 30 ℃, analysis 1 The equivalent percentage of O2 generated.

[0039] The data processing module, which uses a built-in quantitative calculation formula, automatically converts the data based on the detected residual FFA concentration. 1 The O2 generation amount is calculated using the following formula: 1 O2 production equivalent percentage = (initial FFA concentration - remaining FFA concentration) / initial FFA concentration In this formula, the unit of FFA concentration is μmol·L. -1 .

[0040] (4) ROS identification results in Example 1 Fe x S y ROS identification results in mineral systems are as follows Figure 3 As shown. The ROS identification results are as follows: In the control system containing only quartz sand QS (saturated control system QS-Sat. and unsaturated control system QS-Uns.), ·OH (0 μM) was not detected. Figure 3 a) and 1 O2 (FFA indicator showed no change) Figure 3 The generation of (c) was observed, but 89.33%-92.15% of NBT was consumed. Figure 3 (b) This indicates that the QS system contains O2· - The generation of .

[0041] In Fe x S y No detection was found in the QS medium either. 1 O2 generation (FFA indicator unchanged), Figure 3 (c) Compared with the QS control system, Fe x S y &QS saturated environment (83.10%-85.87%) and Fe x S y &QS unsaturated (simulated water level fluctuation) environment (12.53%-62.00%) NBT consumption is significantly reduced. Figure 3 (b) This indicates a significant decrease in O2·- generation. Furthermore, the ·OH concentration showed a trend of first increasing and then decreasing between 24 and 48 h ( Figure 3 in a), Fe x S y Significant ·OH generation was detected in both saturated (up to 6.18 μM) and unsaturated (up to 3.84 μM) environments of the &QS medium, and the ·OH concentration in the saturated environment (2.58-6.18 μM) was consistently higher than that in the unsaturated environment (2.18-3.84 μM).

[0042] (5) ROS identification results in Example 2 Microorganisms-Fe x S y ROS identification results in the "mineral" interaction system are as follows Figure 4 As shown. The ROS identification results are as follows: In a saturated environment, 7.37% of NBT was consumed, while in an unsaturated environment, NBT was almost completely consumed (99%), indicating that more O2 was generated in the unsaturated environment. - .

[0043] Furthermore, the results showed that the concentration of ·OH generated in the saturated environment (4.83 μM) was significantly higher than that in the unsaturated environment (1.55 μM). However, 1 O2 generation showed the opposite trend: the FFA probe reagent decreased by 6.34% in a saturated environment but by 16.89% in an unsaturated environment, indicating that more O2 was generated in the unsaturated system. 1 O2. The above results indicate that changes in the three free radicals in the medium-interaction environment can be accurately detected and quantified.

Claims

1. A high-efficiency synchronous recognition system for ROS combining multiple molecular probes, characterized in that, include: The system comprises a tabletop vertical rotating device, a first reaction device, and a detection device. The first reaction device is light-proof and detachably fixed to the tabletop vertical rotating device, and includes a reaction medium and a contaminant-probe mixed solution. During vertical rotation, the reaction medium of the first reaction device is periodically kept in an alternating wet-expose state to simulate a variable saturation scenario caused by water level fluctuations. The first reaction device includes three parallel and independent contaminant-probe mixed solutions, respectively used for ·OH, O2· - , 1 The detection device is used to quantitatively determine the concentration of stable products or residual probe reagents after each probe reacts with free radicals; the vertical rotation is a rotation on the XZ plane, with its rotation axis parallel to the Y-axis.

2. The high-efficiency synchronous recognition system for ROS using multiple molecular probes as described in claim 1, characterized in that, The system further includes a second reaction device; the first reaction device is light-proof, detachably fixed to a tabletop vertical rotating device, and contains a reaction medium and a contaminant-probe mixed solution; the reaction medium of the second reaction device remains fully submerged during vertical rotation to simulate a saturation scenario; the first and second reaction devices each independently contain three parallel and independent contaminant-probe mixed solutions, respectively for ·OH, O2· - , 1 O2 identification.

3. The high-efficiency synchronous recognition system for ROS using multiple molecular probes according to claim 1 or 2, characterized in that, The probe used for ·OH recognition was benzoic acid (BA), and the product p-hydroxybenzoic acid (p-HBA) was quantitatively determined by high-performance liquid chromatography-ultraviolet detection at 254 nm; for O2· - The identified probe was nitroblue tetrazolium (NBT), and residual NBT was quantitatively determined by UV-Vis spectrophotometry at 260 nm; used for identification. 1 The O2 probe was furfuryl alcohol (FFA) probe, and residual FFA was quantitatively determined by high performance liquid chromatography-ultraviolet detector at 230 nm.

4. The high-efficiency synchronous recognition system for ROS using multiple molecular probes according to claim 3, characterized in that, The concentration of the benzoic acid (BA) probe in the corresponding contaminant-probe mixed solution was 200 mg·L⁻¹. - ¹; The concentration of the nitroblue tetrazolium (NBT) probe in the corresponding contaminant-probe mixture was 0.1 mmol·L⁻¹. - ¹; The concentration of furfuryl alcohol (FFA) probe in the corresponding contaminant-probe mixed solution was 8.5 mmol·L⁻¹. - ¹.

5. The high-efficiency synchronous recognition system for ROS using multiple molecular probes according to claim 1 or 2, characterized in that, The reaction device is a reaction tube containing a reaction medium and a contaminant-probe mixed solution, and the wall of the reaction tube is covered with tin foil to block light.

6. The high-efficiency synchronous recognition system for ROS using multiple molecular probes according to claim 1 or 2, characterized in that, The system also includes a data processing module and an internal displacement calculation formula to automatically calculate the cumulative yield or generation rate of the target free radical based on the residual concentration of the stable product or probe reagent.

7. The high-efficiency synchronous recognition system for ROS using multiple molecular probes according to claim 3, characterized in that, The reaction medium is an iron-sulfur mineral-quartz sand composite; the pollutants in the pollutant-probe mixed solution are PFOA and GenX, and microorganisms may be added or not added selectively.

8. A method for quantitatively detecting ROS using the system according to any one of claims 1–7, comprising the steps of: (1) At room temperature, the reaction apparatus is fixed on a tabletop vertical rotating device and rotated vertically. During the rotation, different probes identify ·OH and O2· respectively. - , 1 O2; (2) After the reaction is complete, collect the liquid sample and use the detection device to quantitatively determine the stable product or residual concentration of the probe reagent after the reaction of each probe with the free radical; (3) Calculate the cumulative yield or generation rate of the target free radical based on the residual concentration of the stable product or probe reagent.

9. The method according to claim 8, characterized in that, In step (1), the table-type vertical rotation device rotates at 20 r·min. -1 Rotation speed: 24 h-48 h.

10. The application of the system according to any one of claims 1-7 in natural / enhanced attenuation ROS detection, advanced oxidation ROS detection and / or ROS optimization in water remediation, and ROS detection in soil remediation.

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