Deuterated zearalenone derivative compounds, preparation method thereof, detection reagent and method for determining content of zearalenone

Abstract

The present application relates to the technical fields of compound synthesis, in particular to a deuterated zearalenone derivative compound, a preparation method thereof, a detection reagent and a method for determining the content of zearalenone, wherein the deuterated zearalenone derivative compound is selected from the compounds shown in the following structural formula: wherein R is selected from any one of C1-C5 alkyl, C1-C5 alkyl-substituted phenyl and a functional group formed by C1-C5 alkyl-substituted biphenyl; and n is any value between 1 and 7. The deuterated zearalenone derivative compound can be used to detect the content of zearalenone, and overcomes the defects of the prior art, such as serious matrix interference caused by isotopic zearalenone, influence on the accuracy of measurement results and low detection efficiency.

Description

Technical Field

[0001] This invention relates to the field of compound synthesis technology, and more specifically, to deuterated zearalenone derivatives, their preparation methods, detection reagents, and methods for determining the content of zearalenone. Background Technology

[0002] Mycotoxins are toxic secondary metabolites produced by fungi on crops or other food sources, contaminating food and feed products and posing a significant threat to human and animal health. Zearalenone (ZEN) is a naturally occurring non-steroidal estrogenic mycotoxin commonly found in crops such as corn, wheat, barley, and sorghum, and their products. It easily accumulates in grains, meat, milk, and eggs. ZEN also accumulates in water and soil, polluting the environment and causing health problems for animals and humans. Furthermore, some studies have shown that ZEN possesses neurotoxicity, immunotoxicity, carcinogenicity, hepatotoxicity, nephrotoxicity, cytotoxicity, and reproductive toxicity. Therefore, monitoring of zearalenone contamination is necessary, including qualitative or quantitative analytical testing.

[0003] Currently, the detection method for zearalenone generally uses the derivative method, but this method is inaccurate. Therefore, isotope detection is used. However, the stable isotope internal standards for zearalenone on the market are mainly fully carbon-labeled zearalenone- 13 C 18 Zearalenone is difficult to synthesize and is generally obtained through biosynthesis, resulting in high synthesis costs. Furthermore, zearalenone with all 18 carbons labeled with 13C has a mass number 18 greater than unlabeled zearalenone, which may lead to mass discrimination in mass spectrometry. Therefore, a new detection reagent is needed that can effectively reduce matrix interference and provide accurate detection results.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] The purpose of this invention is to provide deuterated zearalenone derivative compounds, their preparation methods, detection reagents, and methods for determining the content of zearalenone. This invention provides a novel deuterated zearalenone derivative compound that overcomes the shortcomings of existing technologies where isotopic zearalenone causes severe matrix interference, affecting the accuracy of measurement results and resulting in low detection efficiency.

[0006] This invention is implemented as follows:

[0007] In a first aspect, the present invention provides a deuterated zearalenone derivative compound for detecting the content of zearalenone, wherein the compound is selected from compounds shown in the following structural formula: Wherein, R is selected from any one of the functional groups formed by C1-C5 alkyl, C1-C5 alkyl-substituted phenyl and C1-C5 alkyl-substituted biphenyl; n is any value between 1 and 7.

[0008] Secondly, the present invention provides a method for preparing the deuterated zearalenone derivative compound described in the foregoing embodiments, comprising: mixing deuterated zearalenone with an R-group-containing raw material for reaction, wherein the structural formula of the R-group-containing raw material is as follows: The structural formula of deuterated zearalenone is as follows:

[0009]

[0010] Thirdly, the present invention provides a detection reagent comprising the deuterated zearalenone derivative compound described in the foregoing embodiments.

[0011] Fourthly, the present invention provides a method for determining the content of zearalenone, comprising: pre-treating a sample containing zearalenone and mixing it with a derivatizing reagent to form a derivatized sample containing zearalenone derivative.

[0012] The derivatized sample to be tested is mixed with the deuterated zearalenone derivative compound described in the aforementioned embodiments or the detection reagent for detecting zearalenone content described in the aforementioned embodiments;

[0013] Then, its mass spectrometry was detected, and the content of the zearalenone derivative was determined based on the peak area of ​​the mass spectrometry results;

[0014] The amount of the deuterated zearalenone derivative compound added is not less than 100 μg / L;

[0015] Preferably, the amount of the deuterated zearalenone derivative compound added is 200 μg / L-1000 μg / L.

[0016] The present invention offers the following advantages: This invention provides a novel deuterated zearalenone derivative compound that can be used to detect the content of zearalenone in complex samples. Furthermore, it effectively eliminates matrix interference during the detection process, improving the accuracy and efficiency of the measurement results. Simultaneously, the synthesis method of this deuterated zearalenone derivative compound does not require expensive transition metal catalysts, and offers advantages such as mild reaction conditions, simple synthesis process, good selectivity, environmental friendliness, low cost, high conversion rate, and high product purity, making it suitable for large-scale production. Attached Figure Description

[0017] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 Nuclear magnetic resonance spectra provided for characterization of embodiments of the present invention;

[0019] Figure 2 High-resolution time-of-flight mass spectrometry (Q-TOF) images provided for characterization of embodiments of the present invention;

[0020] Figure 3 High-performance liquid chromatography (HPLC) spectra provided for characterization in embodiments of the present invention. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0022] In a first aspect, embodiments of the present invention provide a deuterated zearalenone derivative compound for detecting the content of zearalenone, which is selected from compounds shown in the following structural formula:

[0023] Wherein, R is selected from any one of the functional groups formed by C1-C5 alkyl, C1-C5 alkyl-substituted phenyl and C1-C5 alkyl-substituted biphenyl; n is any value between 1 and 7.

[0024] For example, R can be a C1-C5 alkyl-substituted phenyl, preferably a C1-C5 alkyl polysubstituted phenyl; more preferably a C1-C5 alkyl disubstituted phenyl or a C1-C5 alkyl trisubstituted phenyl.

[0025] For example, it is selected from compounds with the following structural formula:

[0026] R1 and R2 are each independently selected from C1-C5 alkyl groups, and n is any value between 1 and 7.

[0027] The alkyl groups mentioned above refer to C1-C5 alkyl groups such as methyl, ethyl, propyl, and butyl, and n is any value between 1 and 7, such as 1, 2, 3, 4, 5, 6, and 7.

[0028] Specifically, the deuterated zearalenone derivative is selected from any one of the compounds shown in the following structural formulas:

[0029]

[0030] Where n is any value between 1 and 7, such as 1, 2, 3, 4, 5, 6 and 7.

[0031] Secondly, the present invention provides a method for preparing the deuterated zearalenone derivative compound described in the foregoing embodiments, comprising:

[0032] S1, Synthesis of deuterated zearalenone;

[0033] A mixture of zearalenone, heavy water, and a strong base-weak acid salt is reacted to form deuterated zearalenone, the structural formula of which is shown below:

[0034]

[0035] Specifically, the strong base-weak acid salt in the reaction process is selected from any one or more of potassium carbonate, sodium carbonate, and dipotassium hydrogen phosphate; preferably potassium carbonate; the mass ratio of zearalenone to the strong base-weak acid salt is 1:(5-10), for example, any value between 1:(5-10) such as 1:5, 1:6, 1:7, 1:8, 1:9, and 1:10. The mass ratio of zearalenone to the heavy water is 1-(20:100); for example, any value between 1-(20:100) such as 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, and 1:100. The temperature is room temperature, for example, 10-35°C, more preferably 20-30°C. The organic solvent used in the reaction process includes, but is not limited to, any one or more of tetrahydrofuran, N,N-dimethylformamide, and dimethyl sulfoxide, preferably tetrahydrofuran.

[0036] The temperature and alkali provided in the embodiments of the present invention can not only improve the purity of the product deuterated zearalenone, but also improve its yield.

[0037] After the reaction is complete, post-processing is performed. For example, after the reaction is complete, the reaction system is poured into a beaker, a certain volume of sulfuric acid solution (e.g., 0.2M) is added and stirred thoroughly. Then, it is transferred to a separatory funnel, an extractant is added for extraction, the organic phase is separated and collected, dried, filtered and concentrated to obtain the deuterated gibberellic acid product.

[0038] The volume ratio of the reaction solution to the sulfuric acid solution is 1:30-1:60. The reaction system is neutralized with sulfuric acid to terminate the reaction, increase the yield of the reaction product, and maintain the stability of deuterated zearalenone under acidic conditions.

[0039] The extractant is one or more of dichloromethane, n-hexane, and ethyl acetate, with dichloromethane being preferred.

[0040] S2, Synthesis of deuterated zearalenone derivatives;

[0041] The deuterated zearalenone was reacted with a starting material containing an R group, the structural formula of which is as follows: Specifically, the deuterated zearalenone, the R-group-containing raw material, and the reducing agent are mixed and reacted.

[0042] The molar ratio of deuterated zearalenone to the R-group-containing raw material is 1:(1-5); for example, any value between 1:1, 1:2, 1:3, 1:4, and 1:5. The molar ratio of the reducing agent to the deuterated zearalenone is 1:(5-1); for example, any value between 1:1:2, 1:3, 1:4, and 1:5. The reaction temperature is 10-50℃, and the reaction time is 10-20h.

[0043] If the reaction temperature is too high or too low, it may lead to excessive energy consumption or a decrease in yield or purity.

[0044] The reducing agent is selected from sodium cyanoborohydride, sodium triacetoxyborohydride, and lithium aluminum hydride, preferably sodium cyanoborohydride.

[0045] Further, after the reaction was completed, the organic phase was evaporated under reduced pressure to obtain the crude derivatizing agent product. The crude product was then separated by silica gel column chromatography using a gradient elution with an eluent. The target product fraction was collected in glass test tubes, and the solvent was removed by vacuum filtration to obtain the target product. The eluent was a mixed solution of ethyl acetate and petroleum ether in a volume ratio of 1:3-5.

[0046] In summary, the embodiments of this invention utilize the fungal toxin jugazotoxin ketone as a raw material, and achieve efficient preparation of deuterated jugazotoxin derivatives through a simple reaction process of hydrogen-deuterium exchange and dehydration condensation under alkaline conditions in a room temperature water bath. After column chromatography separation, the product purity can reach 99%. The synthesis method provided by this invention does not require expensive transition metal catalysts and has advantages such as mild reaction conditions, simple synthesis process, good selectivity, environmental friendliness, low cost, high conversion rate, and high product purity. It is suitable for large-scale production and can achieve accurate quantification of jugazotoxin ketones in different matrices, showing broad application prospects.

[0047] Thirdly, the present invention provides a detection reagent comprising the deuterated zearalenone derivative compound described in the foregoing embodiments. This detection reagent may also include other excipients or solvents.

[0048] Fourthly, the present invention provides a method for determining the content of zearalenone, comprising: pretreating a sample containing zearalenone and then mixing it with a derivatizing reagent for derivatization, wherein the derivatizing reagent can be... For example, 2-[(3,4-dimethylphenyl)amino]acetylhydrazine can be chosen, and the derivatization conditions can be referred to existing reactions, which will not be detailed in the embodiments of the present invention. The choices of n and R are also the same as described above.

[0049] It should be noted that the sample to be tested containing zearalenone can be a pretreated sample. The pretreatment steps include: adding an organic solvent such as methanol for shaking extraction, centrifuging to collect the supernatant, adding hexane or similar solvent to further remove impurities such as proteins, centrifuging again for purification; transferring the above solution to a solution containing C 18 The solution was purified by shaking and centrifugation in centrifuge tubes containing PSA and anhydrous magnesium sulfate, and the supernatant was collected.

[0050] Next, a derivatizing agent and sodium cyanoborocyanide are added for derivatization.

[0051] The derivatized sample to be tested is then mixed with the deuterated zearalenone derivative compound or detection reagent described in the aforementioned embodiments, and its mass spectrometry is then measured. The content of the zearalenone derivative is determined based on the peak area of ​​the mass spectrometry results. Since the amount of deuterated zearalenone derivative compound added is known and defined, the content of the zearalenone derivative compound can be determined by the ratio of the peak area of ​​the deuterated zearalenone derivative compound to the peak area of ​​the zearalenone-derived compound, and thus the content of zearalenone can be deduced.

[0052] The amount of the deuterated zearalenone derivative compound added is not less than 100 μg / L; the amount of the deuterated zearalenone derivative compound added is 200 μg / L-1000 μg / L.

[0053] It should be noted that the derivatizing agent for zearalenone is determined based on the R-group-containing raw material of deuterated zearalenone.

[0054] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0055] Example 1

[0056] This invention provides a structural formula for a deuterated zearalenone-derived compound:

[0057]

[0058] This invention provides a method for preparing deuterated zearalenone-derived compounds, comprising:

[0059] S1, Synthesis of deuterated zearalenone;

[0060] Perform the synthesis according to the following synthesis path:

[0061] Specifically, 500 mg of zearalenone, 30 mL of heavy water, 6.2 mL of THF, and 3.9 g of potassium carbonate were added sequentially to a 50 mL round-bottom flask. The mixture was stirred at room temperature (25 °C) and reacted overnight. After the reaction was complete, the reaction mixture was poured into a beaker, and 1800 mL of 0.2 M sulfuric acid was added and stirred. The solution was poured into a separatory funnel and extracted three times with 600 mL of dichloromethane. The organic phase was collected, dried over anhydrous magnesium sulfate, filtered, and the dichloromethane was removed under reduced pressure to obtain 456 mg of white crystalline deuterated zearalenone, with a yield of 90% and a purity of 99%.

[0062] S2, Synthesis of deuterated zearalenone derivatives;

[0063] Perform the synthesis according to the following synthesis path:

[0064]

[0065] Specifically, 500 mg of deuterated zearalenone and 305 mg of 2-[(3,4-dimethylphenyl)amino]acetylhydrazine were dissolved in 10 mL of methanol (pH 6.5, pH adjusted with 0.1 M hydrochloric acid standard solution), and 97 mg of sodium cyanoborohydride was added. The reaction was carried out overnight at room temperature (25 °C). After the reaction was completed, the methanol was evaporated to dryness. The crude product was separated by chromatography using a 100-mesh silica gel column, wet-packed, wet-loaded, and purified by gradient elution with ethyl acetate / petroleum ether (1 / 5-1 / 3). The target product fraction was collected, and after removing the solvent under reduced pressure, 462 mg of white deuterated zearalenone derivative compound was obtained, with a yield of 60% and a purity of 99%.

[0066] Characterization

[0067] The zearalenone, deuterated zearalenone, and deuterated zearalenone derivatives of Example 1 were characterized by 1H NMR spectroscopy, high-resolution time-of-flight mass spectrometry, and high-performance liquid chromatography-mass spectrometry. The results are shown in [reference needed]. Figures 1-3 .

[0068] The reaction products were analyzed by ion scanning using liquid chromatography-mass spectrometry (LC-MS). The determination procedure was as follows: 1 mg of zearalenone, the deuterated zearalenone from Example 1, and the deuterated zearalenone derivative were weighed and dissolved in 1 ml of chromatographic-grade methanol to prepare 1000 mg / L stock solutions of zearalenone and deuterated zearalenone. Working solutions were then serially diluted with methanol to 1 mg / L for LC-MS analysis.

[0069] in, Figure 1 The images show the 1H NMR spectra of zearalenone and deuterated zearalenone. Figure 2 High-resolution time-of-flight mass spectrometry (Q-TOF) of deuterated zearalenone; Figure 3 This shows the high-performance liquid chromatography (HPLC) spectrum of deuterated zearalenone derivatives.

[0070] according to Figures 1-3 It can be seen that after the deuteration reaction, the hydrogen ion content of deuterated zearalenone at chemical shift value 6.41 is significantly reduced, as it is replaced by a deuterium atom, resulting in hydrogen-deuterium exchange. The proton NMR spectrum is shown below. Figure 1 As shown, the main chemical shift peaks were assigned as follows: the doublet at 1.38°C corresponds to hydrogen production at 11′; the multiplet at 1.49°C corresponds to hydrogen production at 4′; the multiplet at 1.65°C corresponds to hydrogen production at 9′; the multiplet at 1.80°C corresponds to hydrogen production at 8′; the multiplets from 2.05°C to 2.28°C correspond to hydrogen production at 3′, 4′, 5′, and 7′; the multiplet at 2.40°C corresponds to hydrogen production at 3′; the broad doublet at 2.61°C corresponds to hydrogen production at 7′; the broad doublet at 2.86°C corresponds to hydrogen production at 5′; the multiplet at 5.0°C corresponds to hydrogen production at 10′; the multiplet at 5.68°C corresponds to hydrogen production at 2′; the singlet at 6.35°C corresponds to hydrogen production at 3′; and the two doublets at 7.02°C correspond to hydrogen production at 1′. These results indicate that the chemical shift values ​​correspond to zearalenone and its deuterated product, proving the successful synthesis of deuterated zearalenone.

[0071] according to Figure 2 It can be seen that due to the deuteration reaction, the molecular ion peak of the compound changed in the negative ion mode. The molecular weight of the deuterated gibberellenone is 321.1623, which is basically consistent with the theoretical prediction of 321.1691. This indicates that the conversion of gibberellenone to deuterated gibberellenone was achieved through the deuteration reaction.

[0072] according to Figure 3 It is known that the deuterated zearalenone derivative is in positive ion mode. Furthermore, the theoretical molecular mass of the derivative obtained by reacting deuterated zearalenone with 2-[(3,4-dimethylphenyl)amino]acetylhydrazine is 498.1, while its molecular mass in positive ion mode is 499.1. This is consistent with the actual measured values ​​from the liquid chromatography-mass spectrometry (LC-MS) spectra. Figure 1This invention demonstrates that the desired deuterated zearalenone derivative compound was synthesized.

[0073] Application examples

[0074] The deuterated zearalenone derivative compound provided in Example 1 of this invention was used for detection, and the specific process is as follows:

[0075] Take 2 mL of blank milk sample and place it in a 50 mL centrifuge tube. Add 120 μL of ZON (gibberellenone) (10 ppm), shake thoroughly to mix, then add 10 mL of methanol (pH 6.5), shake for 10 min, centrifuge at 12000 rpm for 10 min, and collect the supernatant in a 50 mL centrifuge tube. Add 10 mL of n-hexane, shake for 10 min, centrifuge at 12000 rpm for 10 min, discard the upper n-hexane layer, and allow to purify. Transfer the above solution to a centrifuge tube containing 80 mg C18, 60 mg PSA, and 80 mg anhydrous magnesium sulfate, shake for 10 min, centrifuge at 12000 rpm for 10 min, and collect the supernatant. Add 1 mL of derivatizing agent (5 ppm), 120 μL of the deuterated zearalenone derivative compound from Example 1 (10 ppm), and 10 mg of sodium cyanoborohydride to the supernatant. Derivatize at 40 °C for 15 hours, dry under nitrogen at 40 °C, reconstitute with 1 mL of the initial mobile phase, filter through a membrane, and determine the recovery rate of zearalenone in the milk sample.

[0076] The testing was conducted using the national standard method, and the specific process is as follows:

[0077] Take 2 mL of blank milk sample and place it in a 50 mL centrifuge tube. Add 120 μL of ZON (10 ppm), shake thoroughly to mix, add 15 mL of anhydrous diethyl ether, and extract by shaking for 5 min. Centrifuge at 4000 rpm for 2 min, transfer the supernatant to a concentration flask, and concentrate to near dryness at below 40 °C. Add 1 mL of chloroform to dissolve the residue, sonicate for 2 min, and transfer to a 10 mL centrifuge tube. Rinse the concentration flask with 3 mL of 0.5 mol / L sodium hydroxide solution, transfer to the same centrifuge tube, vortex to mix, and centrifuge at 4000 rpm for 2 min. Aspirate the upper sodium hydroxide solution. Add 1 mL of phosphoric acid-water solution, mix well, and allow to purify. Elute with 5 mL of water and 5 mL of methanol-water solution, discarding the eluent; then elute with 10 mL of methanol, collecting the eluent. Dry the eluent under nitrogen gas at below 40 °C. The residue was dissolved in 1.0 mL of acetonitrile, vortexed, and filtered through a 0.2 μm microporous membrane. At the same time, standard curves of different concentrations of matrix were prepared, and quantification was performed by external standard method. The recovery rate of gibberellic acid in the milk sample was found to be 92%.

[0078] The detection was performed using the deuterated zearalenone provided in Example 1 of this invention, and the specific process is as follows:

[0079] Take 2 mL of blank milk sample and place it in a 50 mL centrifuge tube. Add 10 mL of methanol (pH 6.5), 120 μL of ZON (10 ppm), and 120 μL of deuterated zearalenone (10 ppm) from Example 1. Shake for 10 min, centrifuge at 12000 rpm for 10 min, and collect the supernatant in a 50 mL centrifuge tube. Add 10 mL of n-hexane, shake for 10 min, centrifuge at 12000 rpm for 10 min, discard the upper n-hexane layer, and allow to purify. Transfer the above solution to a centrifuge tube containing 80 mg C18, 60 mg PSA, and 80 mg anhydrous magnesium sulfate. Shake for 10 min, centrifuge at 12000 rpm for 10 min, and collect the supernatant. Dry under nitrogen at 40 °C, reconstitute with 1 mL of the initial mobile phase, filter through a membrane, and determine the recovery rate of zearalenone in the milk sample. The recovery rate was found to be 89%.

[0080] The recovery rate was calculated as the ratio of the actual detected juvenile ketone content to the actual juvenile ketone content added to the sample. This recovery rate indicates the accuracy of the corresponding detection method in actual samples. Based on the results, it can be seen that using synthesized deuterated juvenile ketone derivatives as internal standards can effectively reduce matrix interference and improve the accuracy of actual sample detection, providing a new approach for the detection of juvenile ketone in actual samples.

[0081] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A deuterated zearalenone derivative compound, characterized in that, The deuterated zearalenone is used for detecting the content of zearalenone, and is selected from the compounds shown in the following structural formula: wherein R is selected from any one of the functional groups formed by C1-C5 alkyl substituted phenyl and C1-C5 alkyl substituted biphenyl; n is any number between 1-7.

2. The deuterated zearalenone derivative compound according to claim 1, characterized in that, R is selected from C1-C5 alkyl multi-substituted phenyl.

3. The deuterated zearalenone derivative compound according to claim 1, characterized in that, R is C1-C5 alkyl di-substituted phenyl or C1-C5 alkyl tri-substituted phenyl.

4. The deuterated zearalenone derivative compound according to any one of claims 1-3, characterized in that, The deuterated zearalenone is used for detecting the content of zearalenone, and is selected from the compounds shown in the following structural formula: R1and R2are each independently selected from the group consisting of C1-C5alkyl, and n is any number between 1 and 7.

5. The deuterated zearalenone derivative compound according to claim 4, characterized in that, R1 and R2 are independently selected from C1-C3 alkyl.

6. The deuterated zearalenone derivative compound according to claim 4, characterized in that, n is any number between 1-5.

7. The deuterated zearalenone derivative compound according to claim 4, characterized in that, n is 1-3.

8. The deuterated zearalenone derivative compound according to any one of claims 1-3, characterized in that, The deuterated zearalenone is used for detecting the content of zearalenone, and is selected from any one of the compounds shown in the following structural formula: , and wherein n is any number between 1-7.

9. A method of preparing the deuterated zearalenone derivative compound according to claim 1, characterized by, The method comprises the following steps: The deuterated zearalenone is mixed with a raw material containing an R group for reaction, wherein the raw material containing the R group has the following structural formula: The deuterated zearalenone has the following structural formula: 。 10. The production method according to claim 9, characterized by, The method comprises the following steps: The deuterated zearalenone, the raw material containing R group and a reducing agent are mixed for reaction.

11. The method of claim 10, wherein, The molar ratio of the deuterated zearalenone to the raw material containing R group is 1: (1-5); The molar ratio of the reducing agent to the deuterated zearalenone is 1: (5-1); The reaction temperature is 10-50℃, and the reaction time is 10-20h; The reducing agent is selected from any one of sodium cyanoborohydride, sodium triacetoxyborohydride and lithium tetrahydroaluminate.

12. The method of claim 10, wherein, The preparation method of the deuterated zearalenone is as follows: zearalenone, heavy water and a strong base weak acid salt are mixed for reaction.

13. The preparation method of claim 12, wherein, The strong base weak acid salt is selected from any one or more of potassium carbonate, sodium carbonate and dipotassium hydrogen phosphate; The reaction temperature is 10-35℃; The mass ratio of the zearalenone to the heavy water is 1- (20:100); The mass ratio of the zearalenone to the strong base weak acid salt is 1: (5-10).

14. An assay reagent, characterized in that, The method comprises the following steps:

15. A method for measuring the content of zearalenone, characterized by, The method comprises the following steps: The sample containing zearalenone to be detected is pretreated, and then mixed with a derivatization reagent to form a sample to be detected containing zearalenone derivative by derivatization; the determination method is not for the purpose of diagnosis and treatment of diseases; The sample to be detected is mixed with the deuterated zearalenone derivative compound of claim 1 or the detection reagent of claim 9, and then the mass spectrum is detected, and the content of the zearalenone derivative is determined according to the peak area of the mass spectrum result; The addition amount of the deuterated zearalenone derivative compound is not less than 100 μg / L.

16. The method for determining the content of zearalenone according to claim 15, characterized in that, The addition amount of the deuterated zearalenone derivative compound is 200 μg / L-1000 μg / L.

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