Application of Fe3O4 (at) MON-NH2-OH in detection of zearalenone

Abstract

The invention belongs to the technical field of fungus detection, and particularly relates to application of Fe3O4 (at) MON-NH2-OH in detection of zearalenone. According to the method, the microporous organic network Fe3O4 (at) MON-NH2-OH is used as an adsorbent for detecting the zearalenone, the extraction efficiency is high, the enrichment effect is good, and the method is suitable for detecting the zearalenone in agricultural products, feed and traditional Chinese medicinal materials and particularly can be used for detecting trace zearalenone in the agricultural products, the feed and the traditional Chinese medicinal materials. The invention further provides a detection method of zearalenone, and the method is simple and convenient to operate, high in recovery rate and capable of being used for detecting trace zearalenone in complex traditional Chinese medicinal materials.

Description

Technical Field

[0001] The present invention belongs to the technical field of zearalenone, and specifically relates to the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Background Art

[0002] During the processes of planting, processing, harvesting, storing, and transporting traditional Chinese medicinal materials, a large number of fungi are likely to grow and toxins accumulate due to improper methods. Zearalenone (ZEN), also known as F-2 toxin, is a mycotoxin mainly produced by fungi of the genus Fusarium such as Fusarium graminearum, Fusarium tricinctum, and Fusarium culmorum. It is commonly present in moldy agricultural products, feeds, and traditional Chinese medicinal materials, and has strong reproductive toxicity, immunotoxicity, and cytotoxicity. Even remaining in the body, it endangers human health. Therefore, accurately detecting the content of zearalenone in agricultural products, feeds, and traditional Chinese medicinal materials is of great significance, and efficient sample pretreatment methods and sensitive detection techniques are equally important. Especially for complex matrices, sufficient enrichment of trace zearalenone is a prerequisite for accurate detection. Traditional sample pretreatment methods such as liquid-liquid extraction, ultrasonic extraction, solid-phase extraction, and immunoaffinity extraction have disadvantages such as complex operation, low recovery rate, long time consumption, and poor stability. Therefore, it is very necessary to establish a simple, fast, and high-recovery pretreatment technology. Summary of the Invention

[0003] In view of the above problems, the present invention provides the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. The present invention uses Fe 3 O 4 @MON-NH 2 -OH as an adsorbent to enrich zearalenone in the analyte, with simple operation and high recovery rate, and can be used for the detection of zearalenone in agricultural products, feeds, and traditional Chinese medicinal materials.

[0004] To achieve the above invention purpose, the present invention adopts the following technical solutions:

[0005] In the first aspect, the present invention provides the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone.

[0006] The present invention utilizes the microporous organic network Fe 3 O 4 @MON-NH 2-OH is used as an adsorbent to enrich zearalenone in the analyte, with high extraction efficiency and good enrichment effect, providing a new method for the detection of zearalenone in agricultural products, feeds and traditional Chinese medicines, especially for the detection of trace zearalenone.

[0007] Combined with the first aspect, the Fe 3 O 4 @MON-NH 2 -OH is prepared by the following method:

[0008] Mix ferric chloride hexahydrate (FeCl 3 ·6H 2 O), sodium acetate and sodium poly(4-styrenesulfonic acid-co-maleic acid) (PSSMA), dissolve them in ethylene glycol (EG), store at 190 - 210 °C for 8 - 72 h, collect the black product with a magnet, wash and dry to obtain Fe 3 O 4 magnetic nanospheres;

[0009] Disperse the Fe 3 O 4 magnetic nanospheres in cuprous iodide (CuI), bis(triphenylphosphine)palladium dichloride (pd(pph 3 ) 2 Cl 2 ), triethylamine and toluene, add 2,5-diethynylterephthalic diamine (DTBA) and 2,4,6-tribromophenol (TBP) after mixing, stir at room temperature, collect the black particles with the assistance of an external magnetic field, perform Soxhlet extraction with methanol and N,N-dimethylformamide (DMF), and then dry the obtained extract to obtain it.

[0010] In the second aspect, the present invention provides a method for detecting zearalenone, which specifically includes the following steps:

[0011] S1. Extract the analyte with an acetonitrile-water mixture to obtain an extract;

[0012] S2. Add Fe 3 O 4 @MON-NH 2 -OH to the obtained extract, perform ultrasonic treatment and then let it stand, then adsorb with a magnet for solid-liquid separation, add a desorbing solution to the obtained solid phase, mix well, perform ultrasonic elution, and collect the eluate;

[0013] S3. Filter the obtained eluate and then detect zearalenone.

[0014] This method first retains zearalenone on Fe 3 O 4 @MON-NH2 -OH, and then the Fe adsorbed with zearalenone is separated from the solution by applying an external magnetic field. After elution, the enriched zearalenone can be obtained. This method is easy to operate and has a high recovery rate, and can be used for the efficient and accurate detection of zearalenone in agricultural products, feeds, traditional Chinese medicines and other substances to be tested. 3 O 4 @MON-NH 2 -OH. This method does not require adjusting the pH value of the solution and is not affected by the ionic strength in the solution, and has good applicability.

[0015] Combined with the second aspect, the volume ratio of acetonitrile to water in the acetonitrile-water mixture described in S1 is (60-90):(40-10). Preferably, 86:14 is adopted.

[0016] Preferably, the extraction method is ultrasonic extraction.

[0017] Combined with the second aspect, the addition amount of the Fe

[0018] O 3 O 4 @MON-NH 2 -OH described in S2 is not less than 3‰ of the mass of the substance to be tested. Preferably, 5‰ is adopted.

[0019] Combined with the second aspect, the time of the ultrasonic treatment described in S2 is 3-15 min.

[0020] Combined with the second aspect, the eluent described in S2 is methanol, ethanol, or methanol or ethanol containing 0.8%-1.2% v / v formic acid. Preferably, methanol is adopted to obtain a better desorption effect.

[0021] Preferably, the ultrasonic elution is at least 3 times. Each time during elution, 0.1-0.5 mL of methanol is used per milligram of Fe 3 O 4 @MON-NH 2 -OH.

[0022] Preferably, the ultrasonic elution is at least 3 times, 1-10 min each time.

[0023] Combined with the second aspect, the filtration described in S3 is filtration with an organic filter membrane with a pore size of 0.22 μm.

[0024] Combined with the second aspect, the detection method described in S3 is high performance liquid chromatography, and the chromatographic conditions include:

[0025] The chromatographic column is a C18 chromatographic column;

[0026] The column temperature is 27-33 °C;

[0027] The detection wavelength is 231 - 241 nm;

[0028] The flow rate is 0.2 - 0.4 mL / min;

[0029] Mobile phase A is an aqueous solution of 0.09 - 0.11% v / v formic acid, and mobile phase B is acetonitrile. Isocratic elution is performed, and the volume percentage of mobile phase B is 31% - 36%.

[0030] Further preferably, the chromatographic column is an ACQUITY UPLC BEH C18 chromatographic column (2.1 mm × 50 mm, 1.7 μm);

[0031] The column temperature is 30 °C;

[0032] The detection wavelength is 236 nm;

[0033] The flow rate is 0.3 mL / min;

[0034] Mobile phase A is an aqueous solution of 0.1% v / v formic acid, and mobile phase B is acetonitrile. Isocratic elution is performed, and the volume percentage of mobile phase B is 36%.

[0035] In a third aspect, the present invention provides the application of the above detection method in detecting zearalenone in traditional Chinese medicinal materials.

[0036] Combined with the third aspect, the traditional Chinese medicinal materials include peach kernels, bitter apricot kernels, coix seeds, semen alpiniae oxyphyllae, earthworms, prunella vulgaris, Chinese dates, nutmeg, dried tangerine peel, and polygala tenuifolia.

[0037] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0038] Compared with the traditional pretreatment method, Fe 3 O 4 @MON-NH 2 -OH obtained by the above preparation method of the present invention has a strong adsorption capacity for zearalenone, and the maximum adsorption capacity is 215.0 mg / g, which is superior to the reported MNGO, Fe 3 O 4 @nSiO 2 @mSiO 2 、Fe 3 O 4 @PDA / MIL-101(Cr), MHNTs@C=C@MIPs, Fe 3 O 4 -MWCNTs-NH 2 、Fe 3 O 4 -HAP@MIPs, PEG-MWCNTs-MNP, Fe 3 O 4 @TAPB-Tp, Fe3 O 4 @MON-NH 2 -OH and other adsorbents. The method for detecting zearalenone provided by the present invention is easy to operate, has a high recovery rate, a low detection limit and good precision, has good applicability, and can be used for the efficient and accurate detection of zearalenone in agricultural products, feeds, traditional Chinese medicines and other substances to be detected. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 It is the process of detecting zearalenone in Example 1 of the present invention;

[0040] Figure 2 It is Fe in Example 1 of the present invention 3 O 4 @MON-NH 2 -OH preparation principle;

[0041] Figure 3 It is Fe in Test Example 1 of the present invention 3 O 4 @MON-NH 2 -OH structure characterization diagram, where Figure a is the thermogravimetric analysis curve, Figure b is the water contact angle test diagram, Figure c is the hysteresis loop diagram, Figure d is the N 2 adsorption-desorption curve, Figure e is the scanning electron microscope diagram, and Figure f is the transmission electron microscope diagram;

[0042] Figure 4 It is zearalenone, 2,5-diethynyl-p-benzenediamine, 2,4,6-tribromophenol in Test Example 1 of the present invention and the Fe prepared in Example 1 3 O 4 magnetic nanospheres and Fe 3 O 4 @MON-NH 2 -OH infrared spectrum;

[0043] Figure 5 It is the influence of different adsorbents on the adsorption effect of zearalenone in Test Example 2 of the present invention;

[0044] Figure 6 It is the influence of the desorbent on the desorption effect of zearalenone in Test Example 3 of the present invention;

[0045] Figure 7 It is the influence of the desorption time on the desorption effect of zearalenone in Test Example 4 of the present invention;

[0046] Figure 8 It is the influence of the desorbent volume on the desorption effect of zearalenone in Test Example 5 of the present invention;

[0047] Figure 9 For the amount of Fe 3 O 4 @MON-NH 2 -OH in Test Example 6 of the present invention on the extraction effect of zearalenone;

[0048] Figure 10 For the addition of Fe in Test Example 7 of the present invention 3 O 4 @MON-NH 2 -OH on the extraction effect of zearalenone under different ultrasonic treatment times;

[0049] Figure 11 For the effect of different pH values on the extraction effect of zearalenone in Test Example 8 of the present invention;

[0050] Figure 12 For the effect of different NaCl concentrations on the extraction effect of zearalenone in Test Example 9 of the present invention;

[0051] Figure 13 For the chromatograms of ten Chinese medicinal materials in Test Example 10 of the present invention;

[0052] Figure 14 For Fe in Test Example 10 of the present invention 3 O 4 @MON-NH 2 -OH Langmuir fitting diagram for adsorbing zearalenone;

[0053] Figure 15 For Fe in Test Example 11 of the present invention 3 O 4 @MON-NH 2 -OH XPS diagrams before and after extracting zearalenone;

[0054] Figure 16 For Fe in Test Example 12 of the present invention 3 O 4 @MON-NH 2 -OH specific extraction effect on zearalenone. Detailed implementation manners

[0055] In order to make the objectives, technical solutions and advantages of the present invention clearer, the following further details the present invention in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0056] Zearalenone is a mycotoxin with strong reproductive toxicity, immunotoxicity, cytotoxicity and even the ability to remain in the body. It is commonly found in moldy agricultural products, feeds and traditional Chinese medicines, posing a threat to human health. Therefore, accurately detecting the content of zearalenone in agricultural products, feeds and traditional Chinese medicines is of great significance, and the sufficient enrichment of zearalenone is a prerequisite for accurate detection. Traditional sample pretreatment methods have disadvantages such as complex operation, low recovery rate, long time consumption and poor stability.

[0057] In view of the above problems, the embodiment of the present invention provides the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Through experimental research, the present invention finds that Fe 3 O 4 @MON-NH 2 -OH, as an adsorbent, enriches zearalenone in the analyte with high extraction efficiency and good enrichment effect, and can overcome the deficiencies of the prior art.

[0058] The embodiment of the present invention also provides a method for detecting zearalenone, which specifically includes the following steps:

[0059] S1. Extract the analyte with an acetonitrile-water mixture to obtain an extract;

[0060] S2. Add Fe 3 O 4 @MON-NH 2 -OH to the obtained extract, perform ultrasonic treatment and then let it stand, and then use a magnet to adsorb for solid-liquid separation. Add a desorbing solution to the obtained solid phase, mix well, perform ultrasonic elution, and collect the eluate;

[0061] S3. Filter the obtained eluate and then detect zearalenone.

[0062] The embodiment of the present invention also provides the application of the above detection method in detecting zearalenone in traditional Chinese medicines.

[0063] The following specifically illustrates the solution of the present invention through specific examples.

[0064] The reagents and drugs used in the following examples:

[0065] Ferric chloride hexahydrate (FeCl 3 ·6H 2O), sodium poly(4-styrenesulfonic acid-co-maleic acid) (PSSMA, Mw = 20000), 1,3,5-tribromobenzene, 2,4,6-tribromophenol (TBP), 2,5-dichlorobenzenesulfonic acid, p-diethynylbenzene were purchased from Aladdin Chemical Co., Ltd. (Shanghai, China). 2,5-Diethynylterephthalamide (DTBA), tetrakis(4-ethynylphenyl)methane, 2,5-dichlorosulfanilic acid, 2,5-dibromoaniline were provided by Zhongkeyanshen Technology Co., Ltd. (Jilin, China). Copper(I) iodide (CuI) was purchased from Sigma-Aldrich (Shanghai, China). Bis(triphenylphosphine)palladium(II) dichloride (Pd(PPh 3 ) 2 Cl 2 ) was provided by Sigma-Aldrich Chemical Technology Co., Ltd. (Shanghai, China). Zearalenone (ZEN) was purchased from Merck Chemical Technology Co., Ltd. (Shanghai, China). Formic acid (FA) was provided by Macklin Biochemical Co., Ltd. (Shanghai, China). N,N-Dimethylformamide (DMF), ethanol (EtOH), ethylene glycol (EG), methanol (MeOH), acetonitrile (ACN), dichloromethane, triethylamine were purchased from Concord Co., Ltd. (Tianjin, China). Sodium hydroxide (NaOH) was provided by JinKe (Tianjin, China), hydrochloric acid (HCl) was provided by Damao Chemical Reagent Factory (Tianjin, China), and toluene was provided by Far East Fine Chemical Co., Ltd. (Shandong, China). 10 kinds of Chinese medicinal materials, peach kernels, bitter apricot kernels, coix seeds, semen myristicae, earthworms, prunella vulgaris, Chinese dates, nutmeg, dried tangerine peel, polygala tenuifolia, were purchased from Bozhou (Anhui, China).

[0066] The main instruments used in the following examples:

[0067] Agilent 1290UHPLC (Agilent Technologies, USA), vibrating sample magnetometer VSM (LDJ Electronics Inc., USA), optical contact angle measuring instrument (Beijing, China), thermogravimetric analyzer TGA (Rigaku, Japan), Fourier transform infrared spectrometer FT-IR (Thermo Scientific, USA), scanning electron microscope SEM (TESCAN, Czech), transmission electron microscope TEM (JEOL, Japan), X-ray photoelectron spectroscopy XPS (Kratos Analytical Ltd, Britain), ultrasonic cleaner (Ningbo Xinzhi Biotechnology Co., Ltd., Zhejiang, China), Vortex-2 vortex mixer (Shanghai HuXi Industrial Co., Ltd., Shanghai, China), low-temperature high-speed centrifuge (Eppendorf, Germany).

[0068] Example 1

[0069] This example provides Fe 3O 4 @MON-NH 2 Application of -OH in detecting zearalenone:

[0070] Step 1: Prepare a standard stock solution of zearalenone. Weigh 1.0 mg of zearalenone standard, add methanol aqueous solution for dissolution, vortex and shake well to prepare a 1.0 mg / mL stock solution, and obtain working solutions with different concentrations by diluting the stock solution with water.

[0071] Step 2: Crush the sample to be tested, weigh 5.00 g and put it into a 50 mL centrifuge tube, add 40 mL of acetonitrile aqueous solution (86:14, v:v) mixture, ultrasonically extract for 20 min, filter the supernatant with filter paper to obtain the extract.

[0072] Step 3: Add 5 mg of Fe 3 O 4 @MON-NH 2 -OH to the extract obtained in Step 2, vortex and mix well, ultrasonically treat for 7 min and then let it stand, use a magnet to adsorb until the adsorbent is separated from the supernatant, discard the supernatant, then add desorbing solution methanol and mix well, ultrasonically elute the loaded zearalenone (repeat elution 3 times, with 0.5 mL of methanol used each time and 5 min of elution time each time, and the total amount of methanol used is 1.5 mL), collect the eluate, filter with a 0.22 μm organic microporous membrane and then concentrate by nitrogen blowing, which can be used for subsequent HPLC detection.

[0073] Step 4: Perform HPLC detection on the working solutions to establish a standard curve; perform HPLC detection on the extract, calculate the concentration of zearalenone in the extract according to the standard curve, and then calculate the content of zearalenone in the sample to be tested.

[0074] The flowcharts of the above Steps 1 to 3 are as Figure 1 shown.

[0075] Fe 3 O 4 @MON-NH 2 -OH is prepared by the following method:

[0076] 1. Preparation of Fe 3 O 4 Magnetic nanospheres

[0077] Mix 0.81 g of FeCl 3 ·6H 2 O, 1.8 g of sodium acetate and 0.7 g of PSSMA, dissolve them in 10 mL of EG, transfer the mixture to a stainless steel autoclave and store it at 190 - 210 °C for 10 h, collect the black product with a magnet, wash it and dry it overnight in vacuum to obtain Fe 3 O4 Magnetic nanospheres

[0078] 2. Fe 3 O 4 @MON-NH 2 -OH Preparation

[0079] Disperse 1.00 g of Fe 3 O 4 magnetic nanospheres in 11.0 mg of CuI, 42.0 mg of pd(pph 3 ) 2 Cl 2 , 30 mL of triethylamine and 30 mL of toluene. After mixing, add 0.23 g of DTBA and 0.33 g of TBP, stir at room temperature, collect the black particulate matter with the assistance of an external magnetic field, then perform Soxhlet extraction with MeOH and DMF, and subsequently dry overnight in vacuo to obtain it. The preparation principle is as Figure 2 shown

[0080] The HPLC detection chromatographic conditions in Step 4 are as follows:

[0081] The chromatographic column is an ACQUITY UPLC BEH C18 chromatographic column (2.1 mm × 50 mm, 1.7 μm);

[0082] The column temperature is 30 °C;

[0083] The detection wavelength is 236 nm;

[0084] The flow rate is 0.3 mL / min;

[0085] Mobile phase A is 0.1% v / v formic acid aqueous solution, mobile phase B is acetonitrile, isocratic elution, and the volume percentage of mobile phase B is 36%;

[0086] The injection volume is 8 μL.

[0087] Examples 2 - 4

[0088] Examples 2 - 4 provide the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Steps 1 - 4 and the preparation method of Fe 3 O 4 @MON-NH 2 -OH are the same as in Example 1, except that the desorbing solution in Step 3 is different, as shown in Table 1 specifically.

[0089] Table 1 Types of Desorbing Solutions for Examples 2 - 4

[0090] Example 2 Example 3 Example 4 Desorbing solution Ethanol Methanol containing 1% v / v formic acid Ethanol containing 1% v / v formic acid

[0091] Examples 5 - 8

[0092] Examples 5 - 8 provide the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Steps 1 - 4 and the preparation method of Fe 3 O 4 @MON-NH 2 -OH are the same as those in Example 1, except that the ultrasonic elution time in Step 3 is different, as shown in Table 2 specifically.

[0093] Table 2 Desorption solution volumes of Examples 5 - 8

[0094] Example 5 Example 6 Example 7 Example 8 Ultrasonic elution time (min) 1 3 7 10

[0095] Examples 9 - 12

[0096] Examples 9 - 12 provide the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Steps 1 - 4 and the preparation method of Fe 3 O 4 @MON-NH 2 -OH are the same as those in Example 1, except that the volume of the desorption solution in Step 3 is different, as shown in Table 3 specifically.

[0097] Table 3 Desorption solution volumes of Examples 9 - 12

[0098] Example 9 Example 10 Example 11 Example 12 Number of elution times 1 2 4 5 Volume of desorbing solution (mL) 0.5 1.0 2.0 2.5

[0099] Examples 13 - 15

[0100] Examples 13 - 15 provide the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Steps 1 - 4 and the preparation method of Fe 3 O 4 @MON-NH 2 -OH are the same as those in Example 1, except that the addition amount of Fe 3 O 4 @MON-NH 2 -OH is different, as shown in Table 4 specifically.

[0101] Table 4 Addition amounts of Fe 3 O 4 @MON-NH 2 -OH addition amounts

[0102] Example 13 Example 14 Example 15 <![CDATA[Fe 3 O 4 @MON-NH 2 -OH addition amount (mg)]]> 3 7 9

[0103] Examples 16 to 19

[0104] Examples 16 to 19 provide Fe 3 O 4 @MON-NH 2 -OH for use in detecting zearalenone. Steps 1 to 4 and the preparation method of Fe 3 O 4 @MON-NH 2 -OH are the same as in Example 1, except that the ultrasonic treatment time after adding Fe 3 O 4 @MON-NH 2 -OH is different, as shown in Table 5 specifically.

[0105] Table 5 Ultrasonic treatment time of Examples 16 to 19

[0106] Example 16 Example 17 Example 18 Example 19 Ultrasonic treatment time (min) 3 5 10 15

[0107] Example 20

[0108] This example provides the application of Fe 3 O 4 @MON-NH 2 -OH in detecting zearalenone:

[0109] Step 1, prepare a zearalenone standard stock solution. Weigh 1.0 mg of zearalenone standard, add methanol aqueous solution for dissolution, vortex and shake well to prepare a 1.0 mg / mL stock solution, and obtain working solutions with different concentrations by diluting the stock solution with water.

[0110] Step 2, pulverize the analyte and weigh 5.00 g into a 50 mL centrifuge tube, add 40 mL of acetonitrile aqueous solution (90:10, v:v) mixture, ultrasonically extract for 20 min, filter the supernatant with filter paper to obtain the extract.

[0111] Step 3, add 5 mg of Fe 3 O 4 @MON-NH 2 -OH to the extract obtained in Step 2, vortex and mix well, ultrasonically treat for 7 min and then let it stand, adsorb with a magnet until the adsorbent is separated from the supernatant, discard the supernatant, then add desorbing liquid methanol and mix well, ultrasonically elute the loaded zearalenone (repeat elution 3 times, each time the methanol dosage is 0.5 mL, each elution time is 5 min, and the total methanol dosage is 1.5 mL), collect the eluate, filter with a 0.22 μm organic microporous filter membrane and then concentrate by nitrogen blowing, which can be used for subsequent HPLC detection. Fe3 O 4 @MON-NH 2 The preparation method of -OH is the same as that in Example 1.

[0112] Step 4: Perform HPLC detection on the working solution to establish a standard curve; perform HPLC detection on the extract, calculate the concentration of zearalenone in the extract according to the standard curve, and then calculate the content of zearalenone in the test substance. The chromatographic conditions for HPLC detection are the same as those in Example 1.

[0113] Example 21

[0114] This example provides Fe 3 O 4 @MON-NH 2 The application of -OH in the detection of zearalenone:

[0115] Step 1: Prepare a standard stock solution of zearalenone. Weigh 1.0 mg of zearalenone standard, add methanol aqueous solution for dissolution, vortex and shake well to prepare a 1.0 mg / mL stock solution, and obtain working solutions with different concentrations by diluting the stock solution with water.

[0116] Step 2: Crush the test substance and weigh 5.00 g into a 50 mL centrifuge tube, add 40 mL of acetonitrile aqueous solution (60:40, v:v) mixture, ultrasonically extract for 20 min, filter the supernatant with filter paper to obtain the extract.

[0117] Step 3: Add 5 mg of Fe 3 O 4 @MON-NH 2 -OH to the extract obtained in Step 2, vortex and mix well, ultrasonically treat for 7 min and then let it stand, adsorb with a magnet until the adsorbent is separated from the supernatant, discard the supernatant, then add desorption solution methanol and mix well, ultrasonically elute the loaded zearalenone (repeat elution 3 times, the amount of methanol used each time is 0.5 mL, the elution time each time is 5 min, and the total amount of methanol used is 1.5 mL), collect the eluate, filter with a 0.22 μm organic microporous filter membrane and concentrate by nitrogen blowing, which can be used for subsequent HPLC detection. The preparation method of Fe 3 O 4 @MON-NH 2 -OH is the same as that in Example 1.

[0118] Step 4: Perform HPLC detection on the working solution to establish a standard curve; perform HPLC detection on the extract, calculate the concentration of zearalenone in the extract according to the standard curve, and then calculate the content of zearalenone in the test substance. The chromatographic conditions for HPLC detection are the same as those in Example 1.

[0119] Comparative Example 1

[0120] This comparative example provides the application of Fe 3 O 4 @MON-NH 2 in the detection of zearalenone. Steps 1 to 3 are the same as in Example 1, where in Step 3, Fe 3 O 4 @MON-NH 2 is used to replace Fe 3 O 4 @MON-NH 2 -OH in Example 1. In this comparative example, Fe 3 O 4 @MON-NH 2 is prepared by the following method:

[0121] 1. Preparation of Fe 3 O 4 magnetic nanospheres: The same as in Example 1.

[0122] 2. Preparation of Fe 3 O 4 @MON-NH 2

[0123] Disperse 1.00 g of Fe 3 O 4 magnetic nanospheres in 11.0 mg of CuI, 42.0 mg of pd(pph 3 ) 2 Cl 2 , 30 mL of triethylamine and 30 mL of toluene. After mixing, add 0.63 g of tetrakis(4-ethynylphenyl)methane and 0.25 g of 2,5-dibromoaniline, stir at room temperature, and collect the black particles with the assistance of an external magnetic field. Then perform Soxhlet extraction with MeOH and DMF, and dry overnight in vacuo to obtain it.

[0124] Comparative Example 2

[0125] This comparative example provides the application of Fe 3 O 4 @MON-NH 2 -SO 3 H in the detection of zearalenone. Steps 1 to 3 are the same as in Example 1, where in Step 3, Fe 3 O 4 @MON-NH 2 -SO 3 H is used to replace Fe 3 O 4 @MON-NH 2 -OH in Example 1. In this comparative example, Fe 3 O 4 ​@MON-NH 2 -SO 3 H is prepared by the following method:

[0126] 1. Fe 3 O 4 Preparation of magnetic nanospheres: The same as in Example 1.

[0127] 2. Fe 3 O 4 @MON-NH 2 -SO 3 Preparation of H

[0128] Disperse 1.00 g of Fe 3 O 4 magnetic nanospheres in 11.0 mg of CuI, 42.0 mg of pd(pph 3 ) 2 Cl 2 , 30 mL of triethylamine and 30 mL of toluene. After mixing, add 0.63 g of tetrakis(4-ethynylphenyl)methane and 0.24 g of 4-amino-2,5-dichlorobenzenesulfonic acid, stir at room temperature, and collect the black particles with the assistance of an external magnetic field. Then perform Soxhlet extraction with MeOH and DMF, and dry overnight in vacuo to obtain the product.

[0129] Comparative Example 3

[0130] This comparative example provides the application of Fe 3 O 4 @MON-OH in the detection of zearalenone. Steps 1 to 3 are the same as in Example 1, where in Step 3, Fe 3 O 4 @MON-OH is used to replace Fe 3 O 4 @MON-NH 2 -OH in Example 1. Fe 3 O 4 @MON-OH in this comparative example is prepared by the following method:

[0131] 1. Fe 3 O 4 Preparation of magnetic nanospheres: The same as in Example 1.

[0132] 2. Preparation of Fe 3 O 4 @MON-OH

[0133] Disperse 1.00 g of Fe 3 O 4 magnetic nanospheres in 11.0 mg of CuI, 42.0 mg of pd(pph 3 )2 Cl 2 In 30 mL of triethylamine and 30 mL of toluene, after mixing, 0.19 g of 1,4 - diethynylbenzene and 0.33 g of TBP were added. Stir at room temperature, and collect the black particles with the assistance of an external magnetic field. Then perform Soxhlet extraction with MeOH and DMF, and dry overnight in vacuo to obtain the product.

[0134] Comparative Example 4

[0135] This comparative example provides the application of Fe 3 O 4 @MON - SO 3 H in the detection of zearalenone. Steps 1 - 3 are the same as in Example 1, where in Step 3, Fe 3 O 4 @MON - SO 3 H is used to replace Fe 3 O 4 @MON - NH 2 -OH in Example 1. Fe 3 O 4 @MON - SO 3 H in this comparative example was prepared by the following method:

[0136] 1. Preparation of Fe 3 O 4 Magnetic nanospheres: The same as in Example 1.

[0137] 2. Preparation of Fe 3 O 4 @MON - SO 3 H

[0138] Disperse 1.00 g of Fe 3 O 4 magnetic nanospheres in 11.0 mg of CuI, 42.0 mg of pd(pph 3 ) 2 Cl 2 、30 mL of triethylamine and 30 mL of toluene. After mixing, 0.63 g of tetrakis(4 - ethynylphenyl)methane and 0.22 g of 2,5 - dichlorobenzenesulfonic acid were added. Stir at room temperature, and collect the black particles with the assistance of an external magnetic field. Then perform Soxhlet extraction with MeOH and DMF, and dry overnight in vacuo to obtain the product.

[0139] Comparative Example 5

[0140] This comparative example provides the application of Fe 3 O 4 @MON - NH 2 -OH in the detection of zearalenone. Steps 1 - 3 and Fe3 O 4 @MON-NH 2 The preparation method of -OH is the same as that of Example 1, except that the desorbing solution in Step 3 is acetonitrile.

[0141] Comparative Example 6

[0142] This comparative example provides the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Steps 1 to 3 and Fe 3 O 4 @MON-NH 2 The preparation method of -OH is the same as that of Example 1, except that the desorbing solution in Step 3 is acetonitrile containing 1% v / v formic acid.

[0143] Comparative Example 7

[0144] This comparative example provides the application of Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone. Steps 1 to 3 and Fe 3 O 4 @MON-NH 2 The preparation method of -OH is the same as that of Example 1, except that the addition amount of Fe 3 O 4 @MON-NH 2 -OH in Step 3 is 1 mg.

[0145] Inspection Example 1

[0146] This inspection example characterized the structures of the magnetic nanospheres of Fe 2 prepared in Example 1 and Fe 3 O 4 by means of TGA, surface water contact angle, hysteresis loop, N 3 O 4 @MON-NH 2 -OH through adsorption - desorption, SEM and TEM.

[0147] (1) The change process of the weight of Fe 3 O 4 @MON-NH 2 -OH with temperature (30 - 700 °C) is as shown in Figure 3 Figure a therein. The curve shows that Fe 3 O 4 @MON-NH 2 -OH remains stable up to 260 °C, proving that it can be applied at room temperature.

[0148] (2) Fe 3 O 4 @MON-NH 2 -OH has a water contact angle of 24°, as shown in Figure 3 Figure b therein, lower than that of Fe 2 without -NH 3 O 4 @MON (147.3°) and higher than that of Fe 3 O 4 magnetic nanospheres (0°), indicating that MON-NH 2 -OH was successfully modified on Fe 3 O 4 and the prepared Fe 3 O 4 @MON-NH 2 -OH has hydrophilicity.

[0149] (3) As shown in Figure 3 Figure c therein, the saturation magnetization (Ms) values of Fe 3 O 4 magnetic nanospheres and Fe 3 O 4 @MON-NH 2 -OH are 53.1 and 40.9 emu g -1 respectively, indicating that it is feasible to use this Fe 3 O 4 @MON-NH 2 -OH as an adsorbent for detecting zearalenone and it can be easily separated from the water matrix with the help of an external magnetic field, demonstrating that the prepared adsorbent has good magnetic separation performance.

[0150] (4) As shown in Figure 3 Figure d therein, Fe 3 O 4 @MON-NH 2 -OH has a higher BET surface area (140.2 m 3 O 4 g 2 -1) than that of Fe -1 O 2 magnetic nanospheres (18.6 m -1 g 2 -1), verifying the successful modification of porous MON-NH 3 O 4 on Fe.

[0151] (5) Figure 3 Figure e therein shows that at room temperature, Fe 3 O 4@MON-NH 2 -OH has a uniform spherical shape and good dispersibility, with a diameter of about 300 nm. Figure 3 The f figure in shows that MON-NH with a thickness of about 25 nm 2 -OH has successfully grown on the surface of Fe 3 O 4 surface.

[0152] This test example also tested zearalenone, 2,5-diethynyl-p-phenylenediamine, 2,4,6-tribromophenol, and the Fe prepared in Example 1 3 O 4 magnetic nanospheres, Fe 3 O 4 @MON-NH 2 -OH, and Fe 3 O 4 @MON-NH 2 -OH@ZEN (the precipitate obtained after "standing still after ultrasonic treatment for 7 minutes" in Step 3) was subjected to infrared detection, as Figure 4 shown. The FT-IR peaks at 2191 and 1619 cm -1 are respectively attributed to the stretching of the typical triple bond of the MON skeleton and the aromatic C═C bond. The FT-IR spectrum of Fe 3 O 4 @MON shows an Fe-O peak at 587 cm -1 , indicating the successful binding of Fe 3 O 4 to MON. The peaks at 3369 and 1490 cm -1 are respectively attributed to the stretching vibrations of -OH and C-N bonds, confirming that Fe 3 O 4 @MON has successfully modified -OH and -NH 2 groups, and the bifunctional MON has been successfully synthesized. Compared with TBP, the -C-Br peak of Fe 3 O 4 @MON disappears at 595 cm -1 , further proving the successful coupling of the halogenated monomer TBP and the alkynyl monomer DTBA. In the FT-IR spectrum of Fe 3 O 4 @MON-NH 2 -OH@ZEN, the peaks at 1690 and 1715 cm -1 are respectively attributed to the typical C═O stretching vibrations of the ketone carbonyl and ester carbonyl in the ZEN molecule, and the peaks at 1223 and 1012 cm -1 are attributed to the C-O stretching vibration of the O-C═O functional group in ZEN, proving that Fe 3 O4 @MON-NH 2 -OH successfully extracted ZEN. In addition, located on Fe 3 O 4 @MON-NH 2 The O-H characteristic peak on -OH shifted from 3369 cm -1 to 3375 cm -1 after the extraction of ZEN, verifying the existence of hydrogen bond interaction between Fe 3 O 4 @MON-NH 2 -OH and ZEN. At the same time, the characteristic peak representing C=C shifted from 1619 cm -1 to 1628 cm -1 , indicating the existence of π-π and hydrophobic interactions between Fe 3 O 4 @MON-NH 2 -OH and ZEN.

[0153] Test Example 2

[0154] This test example examined the adsorption effects of different magnetic adsorbents in Example 1 and Comparative Examples 1-4 on zearalenone under the HPLC detection conditions in Example 1. The results are as Figure 5 shown. The Fe 3 O 4 @MON-NH 2 -OH prepared in Example 1 had the best enrichment effect on zearalenone.

[0155] Test Example 3

[0156] This test example examined the detection results of elution with different eluents in Examples 1-4 and Comparative Examples 5 and 6 under the HPLC detection conditions in Example 1, as Figure 6 shown. The desorption effect of MeOH was the best. The desorption efficiency was relatively low when using acetonitrile or acetonitrile added with 1% FA as the eluent. Relatively ideal desorption efficiency could also be obtained in ethanol or MeOH added with 1% FA, but it was still inferior to methanol.

[0157] Test Example 4

[0158] This test example examined the detection results at different desorption times in Example 1 and Examples 5-8 under the HPLC detection conditions in Example 1, as Figure 7 shown.

[0159] Test Example 5

[0160] This test example examined the detection results at different eluent volumes in Example 1 and Examples 9-12 under the HPLC detection conditions in Example 1, as Figure 8As shown in the figure. As the volume of the elution solvent increased from 0.5 mL to 1.5 mL, the recovery rate of zearalenone increased. However, after further increase, the extraction efficiency did not increase significantly.

[0161] Test Example 6

[0162] In this test example, the detection results under different dosages of Fe 3 O 4 @MON-NH 2 -OH in Examples 1, 13 to 15 and Comparative Example 7 were investigated using the HPLC detection conditions in Example 1, as Figure 9 shown. As the dosage of Fe 3 O 4 @MON-NH 2 -OH increased, the recovery rate of zearalenone increased. However, after 5 mg, the change in the recovery rate of zearalenone was not significant.

[0163] Test Example 7

[0164] In this test example, the detection results under different ultrasonic treatment times after adding Fe 3 O 4 @MON-NH 2 -OH in Examples 1, 16 to 19 were investigated using the HPLC detection conditions in Example 1, as Figure 10 shown.

[0165] Test Example 8

[0166] This test example investigated the effect of pH on the extraction efficiency of zearalenone in the range of pH 3 to 10.

[0167] 1. Sample Preparation

[0168] Measure a certain amount of water in a beaker, and gradually add 0.1 mol / L NaOH or 0.1 mol / L HCl dropwise to make the pH values 3, 4, 5, 6, 7, 8, 9, and 10 respectively. Then, use the aqueous solutions with different pH values above to prepare 0.1 ppm ZEN test solutions.

[0169] 2. Magnetic Solid Phase Extraction (Carry out 3 parallel experiments simultaneously)

[0170] ⑴ Adsorption: Weigh 5 mg of the magnetic adsorbent Fe 3 O 4 @MON-NH 2 -OH prepared in Example 1 into a 50 mL centrifuge tube, measure 50 mL of 0.1 ppm ZEN test solution into the centrifuge tube, vortex and mix well, and then ultrasonicate for 7 min.

[0171] ⑵ Separation: Place the magnet at the bottom of the centrifuge tube to allow all the adsorbent to sink. With the centrifuge tube close to the magnet, pour off the supernatant, and at this time, the adsorbent is retained at the bottom of the centrifuge tube.

[0172] ⑶ Elution: Elute with 0.5 mL of methanol three times, ultrasonicate for 5 min each time, and collect the eluate with the centrifuge tube close to the magnet.

[0173] ⑷ Enrichment: Filter the collected eluate through a 0.22 μm organic filter membrane into a centrifuge tube, blow it to dry with nitrogen to 0.5 mL, centrifuge at 14000 rpm at 4 °C for 10 minutes, and take the supernatant for standby.

[0174] 3. Detection: Perform HPLC detection on the supernatant, and the chromatographic conditions are the same as in Example 1.

[0175] The results showed that as the pH of the test solution increased from 3 to 7, the recovery rate of zearalenone increased and reached the maximum at pH 7. As the pH increased from 7 to 10, the recovery rate of zearalenone decreased, as Figure 11 shown. The reason may be that: the pKa value of zearalenone is 7.58, while the isoelectric point of Fe 3 O 4 @MON-NH 2 -OH is between 6 and 7. When pH < 6, Fe 3 O 4 @MON-NH 2 -OH carries a positive charge, and the zearalenone under study is protonated. Therefore, the electrostatic repulsion between the positive charges leads to a decrease in the extraction efficiency of Fe 3 O 4 @MON-NH 2 -OH for zearalenone. When the solution pH increases to 7, the electrostatic attraction helps to improve the extraction efficiency of Fe 3 O 4 @MON-NH 2 -OH for zearalenone. When pH > 7, zearalenone exists in a dissociated form, and the hydrogen bond interaction between zearalenone and Fe 3 O 4 @MON-NH 2 -OH is disrupted, and electrostatic repulsion appears, resulting in a decrease in the extraction efficiency. As is well known, the pH value of pure water is usually less than 7. Therefore, the pH is not adjusted in the present invention.

[0176] Test Example 9

[0177] This test example investigated the effect of ionic strength on the extraction efficiency of zearalenone in the NaCl concentration range of 0 - 0.1 mol / L.

[0178] 1. Sample preparation

[0179] Take 900 μL of the 0.1 mg / mL ZEN stock solution prepared in Step 1 of Example 1, add 900 mL of water, and prepare a 0.1 ppm test solution to be measured. Use a measuring cylinder to measure 150 mL (6 portions) of the test solution to be measured, add a certain amount of solid NaCl respectively, shake well and dissolve to make the NaCl solution concentrations be 0, 0.02, 0.04, 0.06, 0.08, and 0.10 mol / L respectively.

[0180] 2. Magnetic solid-phase extraction (conduct 3 parallel experiments simultaneously): same as Test Example 8.

[0181] 3. Detection: Perform HPLC detection on the supernatant, and the chromatographic conditions are the same as those in Example 1.

[0182] The results are as Figure 12 shown. On the one hand, due to the existence of the salting-out effect, the addition of salt will reduce the solubility of zearalenone in the aqueous solution. On the other hand, the addition of NaCl will also increase the viscosity and matrix interference of the sample solution, resulting in a decrease in the recovery rate of zearalenone. Therefore, the addition of NaCl is not conducive to the extraction of zearalenone, and the solution ionic strength is not adjusted in the present invention.

[0183] Test Example 10

[0184] This test example evaluated the method of Example 1.

[0185] (1) Evaluation results of linear curve, detection limit, quantification limit, recovery rate, and precision

[0186] In the concentration range of 1 - 2500 μg / L, a calibration curve was plotted with the peak areas obtained by HPLC-DAD analysis. The linear range was 5 - 2500 μg / L, and the correlation coefficient R 2 was 0.9995.

[0187] By reducing the concentration of zearalenone to a signal-to-noise ratio (S / N) of 3 and 10, the detection limit (LOD) and quantification limit (LOQ) were determined to be 3 and 5 μg / L respectively.

[0188] The RSD values of intra-day (n = 6) and inter-day (n = 6) precision at 3 concentration levels were in the ranges of 0.2 - 0.7% and 1.7 - 2.4% respectively, and the between-batch precision range was between 0.7 - 6.0%.

[0189] Ten traditional Chinese medicinal materials such as peach kernels, bitter apricot kernels, coix seeds, semen alpiniae oxyphyllae, earthworms, prunella vulgaris, Chinese dates, nutmeg, tangerine peel, and polygala tenuifolia were selected to detect zearalenone by the method of Example 1 (Table 6). When the spiked concentrations of zearalenone were 10, 25, and 100 μg / L respectively, the corresponding HPLC chromatograms were as Figure 13As shown (in the same order of medicinal materials above). The recoveries of zearalenone in 10 Chinese medicinal materials were between 80.7 - 106.0%. The results showed that the proposed method has great potential and good applicability.

[0190] Table 6 Spike recoveries of ten Chinese medicinal materials (n = 3)

[0191]

[0192] (2) Fe 3 O 4 @MON-NH 2 -OH adsorption capacity test results

[0193] Figure 14 For Fe 3 O 4 @MON-NH 2 -OH Langmuir fitting graph for zearalenone adsorption, parameters are shown in Table 7. The maximum adsorption capacity of Fe 3 O 4 @MON-NH 2 -OH for zearalenone is 215.0 mg / g.

[0194] Langmuir model:

[0195]

[0196] Where C e is the concentration at adsorption equilibrium (mg / L); Q e is the adsorption capacity at equilibrium concentration C e (mg / g); Q m is the maximum adsorption amount (mg / g); k is the Langmuir constant (L / mg)

[0197] Table 7 Parameters after Langmuir fitting

[0198]

[0199] (3) Matrix effect influence

[0200] For the same analysis object, different samples may exhibit different matrix effects. Generally, the matrix effect can be confirmed by comparing the ratio of the measured value of the analyte in the blank matrix solution to the measured value at the same concentration in pure solvent (matrix factor). If the matrix factor is not within the range of 85% - 115%, it is considered that there is a significant matrix effect in the determination. In this experiment, 10 Chinese herbal medicines were used for matrix spiking (zearalenone at 25, 100, 500 μg / L), and the matrix effect was within the acceptable range of 86.5 - 109.1%, as shown in Table 8.

[0201] Table 8 Experimental results of matrix effect

[0202]

[0203] Test Example 11

[0204] This test example provides the adsorption mechanism of Fe 3 O 4 @MON-NH 2 -OH

[0205] In this test example, the spectra of Fe 3 O 4 @MON-NH 2 -OH before and after extracting zearalenone were obtained by XPS analysis, and the results are shown in Figure 15 . As can be seen from the figure, Fe 3 O 4 @MON-NH 2 -OH extracts zearalenone through hydrogen bonds, π-π bonds, and hydrophobic interactions.

[0206] Test Example 12

[0207] To further study the selective enrichment of zearalenone by Fe 3 O 4 @MON-NH 2 -OH, in this test example, aflatoxin G -1 with a concentration of 1000 μg / L, and having similar molecular weight and molecular size, aflatoxin B 2 (AFG 2 ), aflatoxin B 2 (AFB 2 ), aflatoxin B 1 (AFB 1 ) and zearalenone were selected as target analytes, and magnetic solid-phase extraction experiments were carried out under the optimal conditions. The experimental conditions were the same as Steps 3 and 4 in Example 1.

[0208] The results are as shown in Figure 16 , aflatoxin G 2 , aflatoxin B2 Aflatoxin B 1 and zearalenone in Fe 3 O 4 @MON-NH 2 -OH extraction, the recovery rates were approximately 4.4%, 2.8%, 7.8% and 95.1% respectively, indicating that Fe 3 O 4 @MON-NH 2 -OH has excellent selectivity for the extraction of zearalenone.

[0209] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements or improvements made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Fe 3 O 4 @MON-NH 2 -OH in the detection of zearalenone.

2. The application according to claim 1, wherein, The Fe 3 O 4 @MON-NH 2 -OH is prepared by the following method: Mix ferric chloride hexahydrate, sodium acetate and sodium poly(4-styrenesulfonic acid-co-maleic acid), dissolve them in ethylene glycol, store at 190 - 210 °C for 8 - 72 h, collect the black product with a magnet, wash and dry to obtain Fe 3 O 4 magnetic nanospheres; Disperse the Fe 3 O 4 magnetic nanospheres in cuprous iodide, bis(triphenylphosphine)palladium(II) dichloride, triethylamine, and toluene. After mixing, add 2,5-diethynylterephthalic diamine and 2,4,6-tribromophenol, stir at room temperature, and collect the black particulate matter with the assistance of an external magnetic field. Then, perform Soxhlet extraction with methanol and N,N-dimethylformamide, and subsequently dry the obtained extract to obtain the product.

3. A method for detecting zearalenone, wherein, specifically comprising the following steps: S1. Extract the analyte with an acetonitrile - water mixture to obtain an extract; S2. Add Fe 3 O 4 @MON-NH 2 -OH to the obtained extraction solution, let it stand after ultrasonic treatment, then adsorb with a magnet for solid-liquid separation, add a desorbing solution to the obtained solid phase, mix well, elute by ultrasonic wave, and collect the eluate; S3. Filter the obtained eluate and then detect zearalenone.

4. The detection method according to claim 3, wherein, in the acetonitrile - water mixture in S1, the volume ratio of acetonitrile to water is (60 - 90)﹕(40 - 10).

5. The detection method according to claim 3, wherein, The amount of Fe 3 O 4 @MON-NH 2 -OH added is not less than 3‰ of the mass of the analyte; and / or the time of ultrasonic treatment in S2 is 3 - 15 min; and / or the desorbing solution in S2 is methanol, ethanol, or methanol or ethanol containing 0.8% - 1.2% v / v formic acid.

6. The detection method according to claim 5, wherein, the desorbing solution in S2 is methanol.

7. The detection method according to claim 6, wherein, The ultrasonic elution is performed at least 3 times. During each elution, 0.1 to 0.5 mL of methanol is used for every milligram of Fe 3 O 4 @MON-NH 2 -OH; and / or the ultrasonic elution is at least 3 times, 1 - 10 min each time.

8. The detection method according to claim 3, wherein, the filtration in S3 is filtration with an organic filter membrane having a pore size of 0.22 μm; and / or the detection method in S3 is high - performance liquid chromatography, and the chromatographic conditions include: the chromatographic column is a C18 chromatographic column; the column temperature is 27 - 33 °C; the detection wavelength is 231 - 241 nm; the flow rate is 0.2 - 0.4 mL / min; mobile phase A is an aqueous solution of 0.09 - 0.11% v / v formic acid, mobile phase B is acetonitrile, isocratic elution, and the volume percentage of mobile phase B is 31% - 36%.

9. The application of the detection method according to any one of claims 3 - 8 in detecting zearalenone in traditional Chinese medicinal materials.

10. The application according to claim 9, wherein, the traditional Chinese medicinal materials include peach kernels, bitter apricot kernels, coix seeds, semen alpiniae oxyphyllae, earthworms, prunella vulgaris, Chinese dates, nutmeg, dried tangerine peel, and polygala tenuifolia.

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