A method for enriching succinate dehydrogenase inhibitors and applications thereof
The application of magnetic ethylene-ammoniated graphene oxide (TMGO) adsorbent has solved the problem of efficient enrichment of succinate dehydrogenase inhibitors in food samples, simplified the pretreatment steps, and improved detection efficiency and material lifespan.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
- Filing Date
- 2023-09-22
- Publication Date
- 2026-06-12
AI Technical Summary
Existing technologies for rapid food safety testing suffer from low and inadequate sample pretreatment efficiency, particularly in the extraction and impurity removal of succinate dehydrogenase inhibitors, leading to unsatisfactory testing results.
Magnetic ethylene-amylated graphene oxide (TMGO) was used as an adsorbent. After mixing with the sample to be tested, it was subjected to vortex treatment. Succinate dehydrogenase inhibitors were enriched through magnetic enrichment and elution steps, taking advantage of its high efficiency in separation and recovery.
This method enables rapid and efficient enrichment of succinate dehydrogenase inhibitors, simplifies the operation process, reduces the use of organic solvents, allows for the reuse of adsorbent materials, and lowers detection costs and time.
Smart Images
Figure CN117482574B_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of functional materials technology, and more specifically, to a method for enriching succinate dehydrogenase inhibitors and their applications. Background Technology
[0002] In recent years, frequent food safety incidents have drawn increasing attention to food safety and human health, making the detection of various harmful residues a major concern and highlighting the importance of researching rapid detection technologies. Currently, two bottlenecks hindering the development of rapid food safety detection technologies are low sample pretreatment efficiency and insufficient refinement of rapid detection techniques. Achieving food safety requires a variety of accurate, convenient, rapid, and sensitive rapid detection technologies capable of detecting different contaminants. For samples with complex matrices, the key challenge in rapid detection of food contaminants is sample pretreatment. This involves not only extracting the analyte as completely as possible but also removing other coexisting impurities to avoid interference with detection. Therefore, developing simple, effective, time-saving, and labor-saving pretreatment methods is crucial. Thus, improving rapid detection technologies while simultaneously establishing efficient sample pretreatment methods is urgently needed.
[0003] Currently, the most commonly used pretreatment methods include liquid-liquid extraction (LLE), solid-phase extraction (SPE), solid-phase microextraction (SPME), supercritical fluid extraction (SFE), matrix solid-phase extraction, and QuEChERS technology. However, all of these methods have some drawbacks. SPE involves many steps, is time-consuming and labor-intensive, SPME extraction of coated fibers is expensive and has a short service life, while MSPD, although simple to operate, has fewer steps, and saves labor and time, is not easy to automate. SFE requires large-scale instruments and equipment and has high testing costs. At the same time, adsorbent materials also have the following problems: the adsorption of impurities is limited, some adsorbents are expensive, and most adsorbents cannot be recycled.
[0004] Magnetic solid-phase extraction (MSPE) is a solid-phase extraction technique that uses magnetic materials as adsorbents. It achieves efficient separation of the adsorbent from the sample matrix through an external magnetic field. Compared to traditional solid-phase extraction (SPE), MSPE offers advantages such as simple and rapid pretreatment steps, low organic solvent consumption, high target analyte recovery, and minimal sample loss. Therefore, there is an urgent need for a method to elute and recover succinate dehydrogenase inhibitors from complex matrices such as environmental water or food. Summary of the Invention
[0005] The purpose of this disclosure is to provide a method for rapidly detecting or removing succinate dehydrogenase inhibitors as bactericides from complex matrices such as environmental water or food.
[0006] To achieve the above objectives, this disclosure provides a method for enriching succinate dehydrogenase inhibitors, the method comprising the following steps:
[0007] S1. The test sample, which may contain succinate dehydrogenase inhibitors, is mixed with magnetic ethylene-aminated graphene oxide and vortexed to obtain the first mixture.
[0008] S2. The first mixture is subjected to magnetic enrichment treatment to obtain magnetic ethylene-amino-coated graphene oxide adsorbed with succinate dehydrogenase inhibitor.
[0009] S3. The magnetic ethylene-amylated graphene oxide adsorbed with succinate dehydrogenase inhibitor is contacted with the eluent for elution.
[0010] According to this disclosure, the preparation process of the magnetic ethylene-amylated graphene oxide includes:
[0011] A1. Mix an aqueous solution containing ethylene-amylated graphene oxide with a soluble iron salt and heat in a water bath to obtain a second mixture.
[0012] A2. Add ammonia to the second mixture, stir under nitrogen conditions, and then perform magnetic enrichment, washing, and drying to obtain the magnetic ethylene-ammoniated graphene oxide.
[0013] According to this disclosure, the soluble iron salt is a mixture of divalent and trivalent soluble iron salts; the molar ratio of the divalent and trivalent soluble iron salts is 1:(1-3), preferably 1:2; the divalent soluble iron salt is FeCl2·4H2O; and the trivalent soluble iron salt is FeCl3·6H2O.
[0014] According to this disclosure, the weight ratio of the ethylene-aminated graphene oxide to the iron salt is 1:(5-10).
[0015] According to this disclosure, in step A1, the conditions for water bath heating include: a heating temperature of 60-80℃ and a heating time of 0.5-1h.
[0016] In step A2, the weight ratio of the ammonia water to the second mixture is 1:(10-14);
[0017] The washing and drying process includes: washing with acetonitrile and ultrapure water in sequence, and drying at 50-70℃ for 4-6 hours.
[0018] According to this disclosure, the eluent comprises an acetonitrile and / or methanol solution; preferably, the weight ratio of the acetonitrile to the methanol is 1:(1-3).
[0019] According to this disclosure, the succinate dehydrogenase inhibitor includes at least one of the following: methomyl, carboxin, oxycarboxin, sulfadiazine, fluopyram, cyproconazole, fluopyram, fluopyram, fluopyram, pyrimethanil, pyrimethanil, pyrimethanil, pyraclostrobin, fluopyram, fluopyram, benzovindiflupyr, bifenthiophanate-methyl, thifluzamide, and fluopyram hydroxylamine.
[0020] On the other hand, this disclosure provides the application of the method described in the first aspect in the detection or removal of succinate dehydrogenase inhibitors in food or environmental water.
[0021] According to this disclosure, the food includes at least one of grapes, cucumbers, and honey.
[0022] Through the above technical solution, this disclosure provides a method for rapidly enriching succinate dehydrogenase inhibitors and its application in food and environmental water treatment. Using this method to treat succinate dehydrogenase inhibitors in food and environmental water has the following advantages: simple operation and fast separation speed; low use of organic reagents; the magnetic material used for enriching succinate dehydrogenase inhibitors is recyclable and reusable, has good adsorption effect and a long service life; and it can achieve highly efficient enrichment of succinate dehydrogenase inhibitors.
[0023] Other features and advantages of this disclosure will be described in detail in the following detailed description section. Attached Figure Description
[0024] The accompanying drawings are provided to further illustrate the present disclosure and form part of the specification. They are used together with the following detailed description to explain the present disclosure, but do not constitute a limitation thereof. In the drawings:
[0025] Figure 1 This is a SEM characterization image of TMGO prepared in Example 1.
[0026] Figure 2 This is a TEM characterization image of TMGO prepared in Example 1.
[0027] Figure 3 This is a graph showing the magnetic properties of TMGO prepared in Example 1.
[0028] Figure 4 These are the XRD spectra of TGO and TMGO prepared in Example 1.
[0029] Figure 5 The isotherm curves of nitrogen adsorption and desorption of TMGO prepared in Example 1 are shown.
[0030] Figure 6 This is a thermogravimetric analysis (TGA) diagram of TMGO prepared in Example 1. Detailed Implementation
[0031] The following provides a detailed description of specific embodiments of this disclosure. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit this disclosure.
[0032] The first aspect of this disclosure provides a method for enriching succinate dehydrogenase inhibitors, the method comprising the following steps:
[0033] S1. The test sample, which may contain succinate dehydrogenase inhibitors, is mixed with magnetic ethylene-aminated graphene oxide and vortexed to obtain the first mixture.
[0034] S2. The first mixture is subjected to magnetic enrichment treatment to obtain magnetic ethylene-amino-coated graphene oxide adsorbed with succinate dehydrogenase inhibitor.
[0035] S3. The magnetic ethylene-amylated graphene oxide adsorbed with succinate dehydrogenase inhibitor is contacted with the eluent for elution.
[0036] Optionally, the succinate dehydrogenase inhibitor is selected from any one of methomyl, carbendazim, oxycarbendazim, sulfadiazine, fluopyram, fluopyram, cyproconazole, fluopyram, pyrimethanil, isopyram, pyraclostrobin, pyraclostrobin, fluopyram, fluopyram, benzovindiflupyr, bifenthiophanate-methyl, thifluzamide, and fluopyram hydroxylamine.
[0037] Optionally, the preparation process of the magnetic ethylene-aminated graphene oxide includes:
[0038] A1. Mix an aqueous solution containing ethylene-amylated graphene oxide with a soluble iron salt and heat in a water bath to obtain a second mixture.
[0039] A2. Add ammonia to the second mixture, stir under nitrogen conditions, and then perform magnetic enrichment, washing, and drying to obtain the magnetic ethylene-ammoniated graphene oxide.
[0040] Optionally, the soluble iron salt is a mixture of divalent and trivalent soluble iron salts; the molar ratio of the divalent and trivalent soluble iron salts is 1:(1-3); the divalent soluble iron salt is FeCl2·4H2O; and the trivalent soluble iron salt is FeCl3·6H2O.
[0041] Optionally, the weight ratio of the ethylene-aminated graphene oxide to the iron salt is 1:(5-10).
[0042] Optionally, in step A1, the conditions for water bath heating include: a heating temperature of 60-80℃ and a heating time of 0.5-1h;
[0043] Optionally, in step A2, the weight ratio of the ammonia water to the second mixture is 1:(10-14);
[0044] The washing and drying process includes: washing with acetonitrile and ultrapure water in sequence, and drying at 50-70℃ for 4-6 hours.
[0045] Optionally, the eluent comprises an acetonitrile and / or methanol solution; preferably, the weight ratio of acetonitrile to methanol is 1:(1-3).
[0046] The second aspect of this disclosure provides the application of the above method in the detection or removal of succinate dehydrogenase inhibitors in food or environmental water.
[0047] Optionally, the food includes at least one of grapes, cucumbers, and honey.
[0048] The present disclosure is further described in detail below through examples. All raw materials used in the examples are commercially available.
[0049] Example 1
[0050] Preparation of tetraethylenepentaaminomagnetic graphene oxide (TMGO) material.
[0051] TMGO was synthesized via a coprecipitation method: First, 0.2 g of TGO (purchased from Xianfeng Nanotechnology Co., Ltd.) was dispersed in 200 mL of ultrapure water and sonicated for 30 min. FeCl2·4H2O (0.4 g, 0.002 mol) and FeCl3·6H2O (1.08 g, 0.0004 mol) were dissolved in 40 mL of ultrapure water at a molar ratio of 1:2. The ultrapure water containing TGO was mixed with the ultrapure water containing FeCl2·4H2O (0.4 g, 0.002 mol) and FeCl3·6H2O to obtain a mixed solution. This mixed solution was transferred to a three-necked round-bottom flask and heated in a 70°C water bath for 0.75 h to obtain a second mixed solution. Add 20 mL of ammonia (25 wt%) to the second mixture, stir vigorously for 45 min under nitrogen atmosphere, collect TMGO using an external magnetic field, wash with acetonitrile and ultrapure water in sequence, and finally dry the magnetic nanomaterial at 60 °C for 5 h.
[0052] Example 2
[0053] In this embodiment, the material prepared in Example 1 was characterized by SEM and TEM to determine the morphology, surface distribution and particle size of TGMO.
[0054] Figure 1The study showed that magnetic nanoparticles (MNPs) were randomly modified in a spherical shape on the TGO surface. This indicates that Fe3O4 nanoparticles were successfully attached to TGO.
[0055] Figure 2 The black dots show Fe3O4 and the transparent area are several layers of TGO film. Fe3O4 consists of 10-15 nm nanoscale spherical particles. TEM images show that MNPs are aggregated due to magnetic dipole interactions.
[0056] Figure 3 The magnetic properties of TMGO nanoparticles in a magnetic field range of -10000 to 10000 Oe were investigated using a vibrating sample magnetometer (VSM). TMGO exhibited near-zero hysteresis coercivity and a maximum saturation magnetization (Ms) of 63.73 emu / g, indicating that TMGO can be rapidly separated from solution. Furthermore, dispersed TMGO nanoparticles could be rapidly collected (within 30 seconds) in solution under an applied magnetic field, demonstrating that TMGO possesses sufficiently high magnetic saturation for MSPE.
[0057] Figure 4 The XRD patterns of the prepared TGO and TMGO are shown. Furthermore, the XRD pattern of TMGO showed six new peaks, (220), (311), (400), (422), (440), and (511), which can be attributed to Fe3O4. These results indicate that TMGO was successfully formed by depositing Fe3O4 on the TGO surface and possesses a good crystal structure.
[0058] In this embodiment, the specific surface area, pore volume, and pore size of TMGO were determined using nitrogen adsorption-desorption, and calculated using the Barrette-Joyner-Halenda (BJH) method and the Brunauer-Emmett-Teller (BET) method. Figure 5 As can be seen, a hysteresis loop exists under high pressure, which is a typical type IV isotherm, reflecting a special phenomenon of a mesoporous or microporous structure. The results show that the BET specific surface area of TMGO is 113.93 m². 2 / g, pore volume is 0.25cm³ 3 With a pore size of 8.6 nm, the TMGO prepared in this embodiment has a large specific surface area and pore volume, which is beneficial for actual pre-enrichment and improves the efficiency of succinate dehydrogenase inhibitor absorption from complex matrices.
[0059] This embodiment also performed thermogravimetric analysis (TGA) on the material prepared in Example 1. Figure 6The study shows the mass loss of TMGO nanocomposites in three key stages from 20 to 600 °C. In the first stage (<130 °C), there is a slight reduction in mass, which may be attributed to dehydration and residual fluctuations. Overall, these materials exhibit good thermal stability at room temperature. In the second stage, mass gradually decreases as the temperature increases in the range of 130-450 °C, possibly due to the decomposition of oxygen-containing functional groups such as carboxyl and hydroxyl groups. In the third stage (>450 °C), the prepared material experiences weight loss, possibly due to the vaporization and decomposition of the carbon skeleton.
[0060] Example 3
[0061] The succinate dehydrogenase inhibitor used in this disclosure was purchased from Tianjin Alta. Other reagents whose purchase source is not specified can all be obtained through purchase.
[0062] Succinate dehydrogenase inhibitors include: methomyl, carboxin, oxycarboxin, sulfadiazine, fluopyram, cymoxanil, fluopyram, cymoxanil, fluopyram, pyrimethanil, pyrimethanil, pyrimethanil, pyrimethanil, fluopyram, fluopyram, benzovindiflupyr, bifenthiophanate-methyl, thifluzamide, and fluopyram hydroxylamine (standard products, all purchased from Tianjin Alta).
[0063] Mixed standard working solution: A 10.0 mg / L mixed standard solution was prepared by dissolving individual standard solutions (1000.0 mg / L) of 19 pesticides. This mixed standard solution was then serially diluted with acetonitrile to 800, 500, 200, 50, 10, and 5 μg / L. Matrix mixed standard working solution: Matrix mixed standard working solutions of the above concentrations were prepared using blank sample extracts for the preparation of standard working curves. All standard solutions were stored at -20℃ and prepared fresh for each use.
[0064] Pretreatment of food samples: Water, grapes, cucumbers, grape juice, honey, and mixed fruit and vegetable juice were selected as target samples for testing; grape juice and mixed fruit and vegetable juice were obtained from supermarkets; grape and cucumber samples were homogenized using a high-speed blender, and 5 grams of sample were accurately weighed and placed in a 50 mL centrifuge tube. 5 mL of acetonitrile was added, and the mixture was vortexed for 3 minutes. After centrifugation at 4000 rpm for 5 minutes, the supernatant (5 mL) was collected, dried under nitrogen, and then dissolved in 5 mL of water; 10 grams of honey were dissolved in 100 mL of water, and the water sample was obtained from supermarket purified water; before MSPE extraction, the above test samples were stored in the dark at 4℃; before MSPE extraction, the mixed standard working solution was added to the water, grape, cucumber, grape juice, honey, and mixed vegetable juice samples at three concentrations (5.0, 20.0, and 100.0 μg / L) for laboratory spiking tests.
[0065] Magnetic solid-phase extraction enrichment and concentration process: 15 mg of TMGO adsorbent was added to 3 mL of the sample to be tested in a 10 mL glass bottle, and vortexed for 7 min to obtain the first mixed solution; the adsorbent containing succinate dehydrogenase inhibitor in the first mixed solution was magnetically enriched using a magnet, the supernatant was discarded during the extraction stage, and 3 mL of a mixed solution of acetonitrile and methanol (with a weight ratio of 4:6) was added to the glass bottle for elution to desorb, and vortexed for 7 min to achieve efficient enrichment of succinate dehydrogenase inhibitor bactericides. The adsorbent was collected, filtered using a 0.22 μm filter, and then 2 μL of the extract was injected into an UPLC-MS / MS system (instrument model: Waters Xevo TQ-S ultra-high performance liquid chromatography triple quadrupole tandem mass spectrometer) for detection. The linear range obtained was 5-800 μg / L, and the limit of detection was 0.0050-0.0927 μg / L. The recoveries of 19 succinate dehydrogenase inhibitors were 71.2-119.4%, as shown in Table 1.
[0066] The entire experimental process disclosed herein uses only 3 mL of organic solvent, significantly reducing the amount of organic reagents used compared to the traditional SPE method, thus meeting the requirements of green chemistry. Furthermore, the method provided in this disclosure optimizes experimental conditions, eliminating the need for additional equipment such as centrifuges and vortex mixers during pretreatment, making the experimental process simple and easy to operate. The magnetic material used for enriching succinate dehydrogenase inhibitors is recyclable and reusable, exhibiting good adsorption performance and a long service life; it achieves highly efficient enrichment of succinate dehydrogenase inhibitors with a weak matrix effect.
[0067]
[0068]
[0069]
[0070]
[0071]
[0072] The preferred embodiments of this disclosure have been described in detail above. However, this disclosure is not limited to the specific details of the above embodiments. Within the scope of the technical concept of this disclosure, various simple modifications can be made to the technical solutions of this disclosure, and these simple modifications all fall within the protection scope of this disclosure.
[0073] It should also be noted that the various specific technical features described in the above embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, this disclosure will not describe the various possible combinations separately.
[0074] Furthermore, various different embodiments of this disclosure can be combined in any way, as long as they do not violate the spirit of this disclosure, they should also be regarded as the content disclosed in this disclosure.
Claims
1. A method for enriching succinate dehydrogenase inhibitors, characterized in that, The method includes the following steps: S1. The test sample, which may contain succinate dehydrogenase inhibitors, is mixed with magnetic ethylene-aminated graphene oxide and vortexed to obtain the first mixture. S2. The first mixture is subjected to magnetic enrichment treatment to obtain magnetic ethylene-amino-coated graphene oxide adsorbed with succinate dehydrogenase inhibitor. S3. The magnetic ethylene-amino-modified graphene oxide adsorbed with succinate dehydrogenase inhibitor is contacted with the eluent for elution. The preparation process of the magnetic ethylene-aminated graphene oxide includes: A1. Mix an aqueous solution containing ethylene-amylated graphene oxide with a soluble iron salt and heat in a water bath to obtain a second mixture. A2. Add ammonia to the second mixture, stir under nitrogen conditions, and then perform magnetic enrichment, washing, and drying to obtain the magnetic ethylene-ammoniated graphene oxide.
2. The method according to claim 1, wherein, The soluble iron salt is a mixture of divalent and trivalent soluble iron salts; the molar ratio of the divalent and trivalent soluble iron salts is 1:(1-3); the divalent soluble iron salt is FeCl2·4H2O; and the trivalent soluble iron salt is FeCl3·6H2O.
3. The method according to claim 1, wherein, The weight ratio of the ethylene-aminated graphene oxide to the iron salt is 1:(5-10).
4. The method according to claim 1, wherein, In step A1, the conditions for water bath heating include: heating temperature of 60-80℃ and heating time of 0.5-1h.
5. The method according to claim 1, wherein, In step A2, the weight ratio of the ammonia water to the second mixture is 1:(10-14). The washing and drying process includes: washing with acetonitrile and ultrapure water in sequence, and drying at 50-70℃ for 4-6 hours.
6. The method according to claim 1, wherein, The eluent includes an acetonitrile and / or methanol solution; The weight ratio of acetonitrile to methanol is 1:(1-3).
7. The method according to claim 1, wherein, The succinate dehydrogenase inhibitors include at least one of the following: mefenoxam, carboxin, oxycarboxin, sulfadiazine, fluopyram, cyproconazole, fluopyram, fluopyram, fluopyram, pyrimethanil, pyrimethanil, pyrimethanil, pyraclostrobin, fluopyram, fluopyram, benzovindiflupyr, bifenpyraclostrobin, thifluzamide, and fluopyram hydroxylamine.
8. The use of the method according to any one of claims 1-7 in detecting or removing succinate dehydrogenase inhibitors in food or environmental water.
9. The application according to claim 8, wherein, The food includes at least one of grapes, cucumbers, and honey.