Method for quickly identifying and removing hidden mycotoxin D3G

Abstract

The invention discloses a method for rapidly identifying concealed mycotoxin D3G, which comprises the following steps: 1) extracting a sample to be detected, mixing an extracting solution with D3G standard substances with different concentrations, then mixing with an MGO-coated PEI-ALP dispersion solution, and carrying out room temperature reaction; performing magnetic separation, and taking supernate; (2) mixing the obtained supernate with L-ascorbic acid-2-trisodium phosphate, and incubating to generate ascorbic acid; 3) taking a mixed solution obtained after incubation, adding a metal oxide or metal hydroxide-based nanosheet and AuNPs into the mixed solution, and incubating; and 4) extracting color information of a solution system obtained after incubation, and constructing a relationship between the D3G concentration and the color information to realize rapid identification of the D3G. According to the method, the D3G concentration signal can be converted into the color signal, so that D3G in substrates such as food can be identified, and the method has the advantages of low cost, high speed, convenience in operation and high accuracy, and is suitable for popularization and application.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical analysis and detection, and specifically relates to a method for rapid identification and removal of concealed fungal toxin D3G. Background Art

[0002] Deoxynivalenol-3-glucoside (D3G) is a common hidden fungal toxin. When food containing D3G is consumed, it will be hydrolyzed and reduced to the highly toxic prototype deoxynivalenol (DON) by reductase in the human body. DON is a natural trichothecenes toxin that mainly appears in cereals and cereal products. It can cause food poisoning and vomiting, endangering animal and human health. Therefore, in the field of food processing and testing, the timely detection and removal of D3G has become crucial.

[0003] At present, commonly used D3G quantitative detection methods include liquid chromatography-mass spectrometry (LC-MS / MS), gas chromatography-mass spectrometry (GC-MS / MS), etc. Although these methods have high sensitivity and selectivity, the processing process is complicated, the instruments are expensive, and professional personnel are required to operate, which makes it difficult to meet the market demand for rapid detection. Therefore, it is necessary to develop a convenient, economical, and high-speed detection method to meet the needs of rapid on-site analysis and flexible detection anytime and anywhere. Summary of the invention

[0004] The main purpose of the present invention is to provide a method for detecting hidden fungal toxin D3G using a colorimetric sensor based on nanomaterials. The sensor is mainly composed of MGO@PEI-ALP conjugate and gold nanoparticles (AuNPs), which can convert the signal of D3G concentration into a color signal, and then identify D3G in a matrix such as food; it has the advantages of low cost, fast speed, convenient operation and high accuracy, and is suitable for popularization and application.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] A rapid identification method for concealed mycotoxin D3G comprises the following steps:

[0007] 1) Extract the sample to be tested, mix the obtained extract with D3G standard products of different concentrations, and then mix it with magnetic MGO@PEI-ALP dispersion, react at room temperature to free ALP again and restore its activity; perform magnetic separation and take the supernatant;

[0008] 2) mixing the obtained supernatant with trisodium L-ascorbic acid-2-phosphate (AAP), incubating, and generating ascorbic acid (ALP catalyzes AAP to generate ascorbic acid AA);

[0009] 3) taking the mixed solution obtained after incubation, adding metal oxide or metal hydroxide-based nanosheets and AuNPs thereto, and incubating, the metal ions and AuNPs undergo chelation, causing color change;

[0010] 4) Extracting the color information of the solution system obtained after incubation, constructing the relationship between D3G concentration and color information, and realizing rapid identification of D3G.

[0011] In the above scheme, the sample to be tested can be selected from one or more of wolfberry, rice, dry oatmeal, dry corn kernels, highland barley, etc.

[0012] In the above scheme, the concentration of D3G in the D3G standard in step 1) is 0.6-8.0 μg / L.

[0013] In the above scheme, the room temperature reaction time of step 1) is 25 to 35 minutes.

[0014] In the above scheme, the preparation method of the MGO@PEI-ALP dispersion comprises the following steps: mixing MGO (magnetic graphene oxide) and PEI (polyethyleneimine) in water, heating and stirring, centrifuging, washing, dispersing the resulting precipitate (PEI coated with MGO; MGO@PEI) in water, and then mixing with ALP (alkaline phosphatase), and incubating with shaking at room temperature; centrifuging to obtain the supernatant, adding the ALP color developer, and then shaking and mixing evenly to obtain a dispersion containing MGO@PEI-ALP, and measuring their ultraviolet absorption spectra after 15 minutes.

[0015] In the above scheme, the mass ratio of MGO to PEI is 1:350-450.

[0016] In the above solution, the sheet diameter of the magnetic graphene oxide is less than 5 μm.

[0017] Furthermore, the magnetic graphene oxide comprises magnetic Fe 3 O 4 The particle size is 9 to 11 nm.

[0018] In the above scheme, the molecular weight M of the PEI is w It is 24750-25250.

[0019] In the above scheme, the temperature used for heating and stirring is 55-65°C and the time is 10-14h.

[0020] In the above scheme, the mass ratio of ALP to MGO@PEI is 12 to 600:1.

[0021] In the above scheme, the room temperature oscillation frequency is 180-220 rpm, and the incubation time is 0.5-1.5 h.

[0022] In the above scheme, the ALP color developer can be selected from p-nitrophenyl phosphate (pNPP), 5-bromo-4-chloro-3-indolyl phosphate (BCIP) or nitro blue tetrazolium (NBT), etc.; the mass ratio of pNPP to ALP is 1.8 to 3:1.

[0023] In the above scheme, in the solution system of step 2), the mass ratio of D3G, MGO@PEI-ALP and AAP introduced is (0.13-1.8)*10 -3 :1:(1.3~1.5)*10 4 .

[0024] In the above scheme, the incubation temperature in step 2) is 35-40° C. and the time is 25-35 min.

[0025] In the above scheme, the metal oxide-based nanosheet is MnO 2 Nanosheets, metal hydroxide-based nanosheets are Co(OH) 2 Nanosheets, etc.

[0026] Preferably, the nanosheets are made of Co(OH) 2 The nanosheets have a diameter of 90 to 106 nm and a thickness of 7.5 to 8 nm.

[0027] In the above scheme, the average particle size of the AuNPs is 10 to 15 nm, preferably 12 to 14 nm.

[0028] Preferably, the AuNPs have a regular spherical shape.

[0029] In the above scheme, in the reaction system described in step 3), Co(OH) 2 Nanosheets or MnO 2 The concentration of the nanosheets is 0.1 to 0.17 mM, preferably 0.11 to 0.15 mM; the concentration of the nanogold is 2.1 to 2.34 nM.

[0030] In the above scheme, the mass ratio of the introduced metal oxide or metal hydroxide-based nanosheets to MGO@PEI-ALP is 10 to 20:1.

[0031] In the above scheme, the incubation time in step 3) is 4 to 6 minutes, and the temperature is 23 to 27°C.

[0032] In the above scheme, the color change information is a change value Y based on absorbance, RGB value or color change rule; wherein the change value Y based on absorbance=(I 0 -I) / I0 , I 0 I is the absorbance at 521 nm when the reaction system lacks the target substance D3G, and I is the absorbance at 521 nm after the reaction system reacts with the target substance D3G.

[0033] Further, the method of constructing the relationship between D3G concentration and color change information includes:

[0034] 1) By collecting the UV spectrum of the sample, a calibration curve for determining D3G is established. When the D3G concentration is 0.5-3.5 μg, the color change degree (Y) of the entire reaction system and the D3G concentration (X) show a good linear relationship, thereby achieving quantitative detection of the D3G concentration;

[0035] 2) In order to achieve instrument-free detection, a colorimetric method can also be used, that is, taking pictures of the actual samples, extracting the digital information (R / G / B values) of the collected pictures respectively, analyzing the digital matrix in combination with the chemometric method, and constructing a partial least squares regression (PLSR) model to achieve quantitative detection of D3G concentration.

[0036] 3) Place the D3G mixtures of different concentrations in 96-well plates and add 100 μL of H 2 O and mix well. As the concentration of D3G increases, the color of the system gradually changes from pink to blue-purple, and the qualitative analysis of the concentration of D3G can be achieved through color.

[0037] Furthermore, D3G concentrations of 1.0 ug / L, 3.0 ug / L and 7.0 ug / L were selected as experimental concentrations for adding standard recovered materials.

[0038] Using the above test method, MGO@PEI-ALP attracts D3G through the electrostatic effect and can simultaneously achieve effective removal of D3G in the solution system.

[0039] The principle of the present invention is:

[0040] Magnetic graphene oxide (MGO) usually presents negative charge in aqueous solution. The present invention firstly coats and modifies it with polyethyleneimine (PEI), and the obtained MGO@PEI surface has positive charge. Then, alkaline phosphatase (ALP) is coupled to MGO@PEI to inhibit ALP activity and provide competitive substrate for subsequent recognition of target D3G. Meanwhile, the special structure of D3G causes it to release hydrogen ions (H + ), resulting in a large amount of negative charge in the solution. D3G competes for the magnetic graphene oxide sites (MGO@PEI) in MGO@PEI-ALP through electrostatic adsorption, causing ALP to be freed and restored to activity, thereby causing the ALP color developer to develop color (pink). The restored ALP can further catalyze β-Co(OH)2 The nanosheets are reduced to Co 2+ The stability between AuNPs due to electrostatic repulsion is broken by chelating AuNPs, resulting in aggregation color change; the degree of color change of nanogold aggregation caused by different D3G concentrations is different, causing a slight increase in the absorbance of the sensor at 624nm, and a series of color changes (which can be observed by the naked eye and colorimetric detection). According to the acquired color information and combined with chemometric methods, the rapid identification of hidden fungal toxin D3G can be achieved; at the same time, the magnetism of magnetic graphene oxide can be used to effectively remove D3G from the detection system. In addition, the detection system of the present invention has high selectivity for D3G, which can effectively avoid interference from hidden fungal toxins such as ZEN, T-2, HT-2, and DON.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] 1) This invention proposes for the first time to construct a colorimetric sensor based on the charge adsorption principle using MGO@PEI-ALP conjugates and gold nanoparticles, combined with chemometrics, to achieve rapid visual identification and detection of D3G;

[0043] 2) The sensor constructed by the present invention has the advantages of no instrument detection, simple operation, high sensitivity, fast response, etc., and can realize the rapid, efficient and visual identification of hidden fungal toxin D3G;

[0044] 3) Due to the special magnetic properties of MGO, compared with the traditional colorimetric sensing method, this method can also simultaneously remove D3G, providing a new way for the detection and removal of D3G in complex food and drug samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 TEM characterization diagrams of the AuNPs nanomaterials of the present invention in different states; A: the state of AuNPs before adding D3G, B: the state of AuNPs after adding D3G, C: the state of β-Co(OH) in the AuNPs solution before adding D3G 2 Nanosheet state, D: β-Co(OH) in AuNPs solution after adding D3G 2 Nanosheet state;

[0046] Figure 2 The color change results of AuNPs before and after adding D3G;

[0047] Figure 3 The UV spectra, color response results and calibration curves after adding different concentrations of D3G; A: UV spectrum, B: series displacement results of AuNPs; C: color response results; D: calibration curve;

[0048] Figure 4 The reaction results of the fluorescence sensor and different hidden fungal toxins; A: UV spectrum, B: absorbance of AuNPs at 521nm and 624nm;

[0049] Figure 5 The visualized color reaction results of different actual samples based on the aggregation effect of nano-gold as the concentration of D3G changes;

[0050] Figure 6 The correlation diagram of the actual and predicted concentrations of D3G in different matrices based on R / G / B values; A: wolfberry, B: rice, C: highland barley, D: dry oatmeal, and E: dry corn kernels. DETAILED DESCRIPTION

[0051] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described below by way of embodiments, but the present invention is not limited to the scope of the embodiments.

[0052] In the following examples, AuNPs were introduced in the form of AuNPs dispersion. The specific preparation steps were as follows: 100 mL of 1.0 mM HAuCl 4 The solution was heated to boiling and stirred magnetically; then, 1 mL of 400 mM sodium citrate solution was quickly added; the resulting mixed solution was kept boiling and stirred for 10 min, and after cooling to room temperature, an AuNPs dispersion stabilized by sodium citrate was obtained (the average particle size of AuNPs was 13 nm); combined with the absorbance intensity of AuNPs at 520 nm, the concentration of the AuNPs dispersion was calculated to be 6.7 nM.

[0053] β-Co(OH) 2 The preparation steps of the nanosheets are as follows: a mixed aqueous solution of sodium hydroxide and hydrazine hydrate is used as an alkaline precipitant, and the molar ratio of the introduced cobalt chloride, sodium hydroxide, and hydrazine hydrate is 1:3:6; 0.518 mL of alkaline precipitant solution (the concentration of sodium hydroxide is 0.15 mol / L, the concentration of hydrazine hydrate is 0.3 mol / L) is slowly and evenly added to 0.518 mL of cobalt chloride (concentration is 0.05 mol / L) aqueous solution in a fume hood, and under magnetic stirring, the precipitate color gradually changes from blue-green to rose red; then, the suspension is allowed to stand for 12 hours, the supernatant is taken out, and washed with deionized water and anhydrous ethanol three times, and the washed product is dried at 60°C for 4 hours under a nitrogen environment to obtain rose-red β-Co(OH) 2 Powder; tested, synthesized β-Co(OH) 2 The diameter of the nanosheets is 90 to 100 nm.

[0054] The preparation steps of MGO@PEI-ALP nanoconjugates are as follows: 1.0 mL of MGO solution (0.5 mg / mL, solvent is water) was mixed with 20.0 mL of PEI solution (10.0 mg / mL, solvent is water, molecular weight of PEI is 25000), stirred at 60 °C for 12 h, centrifuged and washed three times, and the resulting precipitate was dispersed again in 20.0 mL of H 2 O to obtain a PEI-coated MGO dispersion (MGO@PEI) with a concentration of 0.4 mg / mL, and then diluted to MGO@PEI solutions with concentrations of 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and 5 μg / mL, respectively; then, 3 mL of ALP solution (200 μg / mL) and 10 mL of MGO@PEI solutions with different concentrations were mixed and incubated with shaking at room temperature for 1 h; then, each mixed solution was centrifuged to obtain a supernatant, and finally, 100 μL of 50 mM p-nitrophenyl phosphate solution (ALP color developer) was added to the supernatant to obtain MGO@PEI-ALP nanoconjugate solution.

[0055] Example 1

[0056] A method for rapid identification of hidden fungal toxin D3G, comprising the following steps:

[0057] 1) Take wolfberry, rice, dry oatmeal, dry corn kernels and highland barley samples respectively, pass them through a 20-mesh sieve, and then use 50 mL of ultrapure water to ultrasonically treat them at 100 Hz and 35°C for 30 min, with a solid-liquid ratio of 1 mg:1 mL; then, filter the centrifugal supernatant with a 0.22 μm filter membrane (4,000 r / min, 10 min) to obtain extracts of different samples;

[0058] 2) Add 50 μL of D3G solution of different concentrations (0.6, 1.0, 2.0, 3.0, 4.0, 5.6, 6.6, 7.0 and 8.0 μg / L) to 50 μL of real sample extract, and then add 100 μL of the obtained MGO@PEI-ALP solution (concentration of 2.3 μg / mL) and react at room temperature for 30 min;

[0059] 3) 100 μL of the obtained supernatant was mixed with 100 μL of AAP solution (concentration of 100 mM) and incubated at 37°C for 30 min;

[0060] 4) Take 100 μL of the resulting mixture and add 100 μL Co(OH) 2 Nanosheet dispersion (0.4 mM) and 100 μL AuNPs solution (6.7 nM), wherein the obtained mixed solution system, after incubation for 5 min, the AuNPs mixed solution was obtained; the introduced Co(OH) 2The mass ratio of nanosheets to MGO@PEI-ALP is 16.17:1;

[0061] 5) using a UV-spectrophotometer to obtain the absorption spectrum of the obtained AuNPs mixture and extract the color information, wherein each D3G concentration was tested three times to construct a chemometric relationship between the D3G concentration (X) and the color information (Y); wherein the concentrations of D3G were 0.3 μg / L, 0.5 μg / L, 1 μg / L, 1.5 μg / L, 2.0 μg / L, 2.8 μg / L, 3.3 μg / L, and 3.5 μg / L, and LOD = 3σ / k, where σ is the standard deviation of the blank value and k is the slope of the fitting curve;

[0062] Y=200.86X+0.0079,(R 2 =0.9975)

[0063] Y=(I 0 -I) / I 0

[0064] In the formula, I 0 I is the absorbance at 521 nm of the reaction system lacking the target substance D3G, and I is the absorbance at 521 nm of the reaction system after the reaction with the target substance D3G.

[0065] Figure 1 The TEM characterization images of the nanomaterials obtained under different conditions show that before adding D3G, AuNPs are uniformly distributed in spherical shapes and dispersed evenly. However, after adding D3G, AuNPs agglomerate and the single regular sphere disappears; β-Co(OH) 2 The nanosheets are regular hexagons with a side length of about 75.57 nm. After adding D3G, Co(OH) 2 The morphology is incomplete and the amount is reduced sharply, indicating that Co(OH) 2 Reduced to Co by ascorbic acid (AA) 2+ .

[0066] Figure 2 It can be seen that compared with the single nanogold solution, the addition of D3G, MGO@PEI-AL, AAP, Co(OH) 2 After the nanosheets are formed, the solution system changes from pink to blue-purple.

[0067] Figure 3 The UV spectra and calibration curves obtained in step 5) after adding different concentrations of D3G show that when the D3G concentration range is 0.5-3.5 μg / L, there is a good linear relationship between the color change degree (Y) of the entire reaction system and the D3G concentration (X); the detection limit (LOD) is 0.30 μg / L, which is far below the maximum limit standard (1.0 mg / kg) prescribed by the EU.

[0068] According to the above method and fitting results, wolfberry, rice, dry oatmeal, dry corn kernels and highland barley were used as actual test samples to measure the recovery rate of D3G. Three tests were conducted for each sample. The spike conditions and results are shown in Table 1.

[0069] Table 1 shows that the recovery rate of D3G in actual samples is between 96.29% and 112.3%, indicating that the fluorescence sensing method constructed in the present invention has good practical value in actual samples.

[0070] Table 1 Recovery rate of D3G detected in actual samples

[0071]

[0072] Specificity studies

[0073] High concentrations of other mycotoxins and hidden mycotoxins (ZEN, T-2, HT-2, DON, all at 30.0 μg / L) were added to MGO@PEI-ALP, and AAP, Co(OH) 2 Nanosheets and AuNPs. The selectivity of the fluorescence sensor for detecting D3G was verified by testing the absorbance of MGO@PEI-ALP after reacting with different hidden fungal toxins.

[0074] like Figure 4 As shown in A, when the concentration of D3G is 3.0 μg / L, except for a slight response to its prototype DON, the sensor has almost no response to other substances. Figure 4 B shows the absorbance at two different wavelengths after adding D3G and other competing substances. The blue is the absorbance at 624nm, which represents the degree of color change. It can be seen that there is a significant difference between adding D3G and adding other substances. After adding D3G, the absorbance ratio is the highest, indicating that the morphology of AuNPs has changed and the particle size has increased, which also shows that the sensor has a strong response to D3G. The results show that the sensor has good selectivity for D3G and can effectively identify the hidden fungal toxin D3G.

[0075] Example 2

[0076] Application of real samples

[0077] Nine different concentrations of D3G solutions (D3G concentrations were 0.3, 0.5, 1, 1.5, 2.0, 2.8, 3.3, 3.5, and 4.0 μg / L) were added to the extracts of different real samples, and the MGO@PEI-ALP conjugate, AAP, β-Co(OH) 2, AuNPs were mixed and then dropped into a 96-well plate in equal amounts. A camera was used to take pictures of the actual samples on the well plate, and the pixel information of 10 10×10 pixel pictures of each concentration was extracted using Photoshop software. The pixel image information was converted into a 300×90 digital matrix using MATLAB software, and the above 90 groups of samples were divided into 61 training sets and 29 test sets. The PLSR method was used to extract the color change information of each reaction well. In PLSR, a linear regression model is fitted by projecting the observed variables (X, R / G / B values ​​of the color) and the predictor variables (Y, the concentration of D3G) into the dimensionality reduction space. The selection of the latent variable LV was based on the root mean square error (RMSE) of cross-validation. The accuracy of the PLSR model was evaluated by the corrected root mean square error (RMSEC) and the predicted root mean square error (RMSEP). R 2 c and R 2 p was used to evaluate the coefficient of determination between the actual and predicted concentrations in the training and prediction sets.

[0078] Figure 5 This is the visualization result of the actual sample changing with the D3G concentration. It can be seen that when other conditions are the same, the color change of the same sample is related to the change of D3G concentration.

[0079] The above PLSR quantitative model results are shown in Table 2, Table 3 and Figure 6 , where Table 2 shows the training set and prediction set division results of D3G sample solutions with different concentrations in different actual samples, and Table 3 shows the D3G concentration results of different PLSR matrices based on RGB values. The model parameters show that the PLSR model works well in predicting D3G concentrations in multiple food samples; Figure 6 It shows that there is a good linear relationship between the actual concentration and the predicted concentration of D3G in various real samples.

[0080] Table 2 The training set and prediction set division results of D3G sample solutions with different concentrations in different actual samples

[0081]

[0082]

[0083] Table 3. Results of D3G concentration in different real samples based on R / G / B images and PLSR

[0084]

[0085] The above description is only a preferred embodiment of the present invention and is not intended to limit the scope of implementation of the present invention. That is, any equivalent changes and modifications made according to the content of the patent application scope of the present invention should be the technology of the present invention.

[0086] In addition, the various embodiments of the present invention may be combined in any way, and as long as they do not violate the concept of the present invention, they shall also be regarded as the contents disclosed by the present invention. The present invention is not limited to the above-mentioned embodiments. For ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications are also regarded as within the scope of protection of the present invention. The contents not described in detail in this specification belong to the prior art known to professional and technical personnel in this field.

Claims

1. A method for rapid identification of hidden fungal toxin D3G, characterized in that: The steps include: 1) Extracting the sample to be tested, mixing the obtained extract with D3G standard products of different concentrations, and then mixing with MGO@PEI-ALP dispersion, reacting at room temperature; performing magnetic separation, and taking the supernatant; 2) mixing the obtained supernatant with L-ascorbic acid-2-phosphate trisodium salt, incubating, and generating ascorbic acid; 3) taking the mixed solution obtained after incubation, adding metal oxide or metal hydroxide-based nanosheets and AuNPs thereto, and incubating; 4) Extracting the color information of the solution system obtained after incubation, constructing the relationship between D3G concentration and color information, and realizing rapid identification of D3G.

2. The rapid identification method according to claim 1, characterized in that: The samples to be tested are one or more of wolfberry, rice, dry oatmeal, dry corn kernels, and highland barley.

3. The rapid identification method according to claim 1, characterized in that: The concentration of D3G in the D3G standard in step 1) is 0.6-8.0 μg / L.

4. The rapid identification method according to claim 1, characterized in that: The room temperature reaction time of step 1) is 25 to 35 minutes.

5. The rapid identification method according to claim 1, characterized in that: The preparation method of the MGO@PEI-ALP dispersion comprises the following steps: mixing MGO and PEI in water, heating and stirring, dispersing the obtained precipitate MGO@PEI in water, then mixing with ALP, incubating with shaking at room temperature, centrifuging to obtain the supernatant, adding the ALP color developer, and shaking and mixing evenly to obtain the MGO@PEI-ALP dispersion.

6. The rapid identification method according to claim 5, characterized in that: The mass ratio of MGO to PEI is 1:350-450.

7. The rapid identification method according to claim 1, characterized in that: The metal oxide-based nanosheets are MnO2 nanosheets, and the metal hydroxide-based nanosheets are Co(OH)2 nanosheets.

8. The rapid identification method according to claim 1, characterized in that: The mass ratio of the introduced metal oxide or metal hydroxide-based nanosheets to MGO@PEI-ALP is 10 to 20:

1.

9. The rapid identification method according to claim 1, characterized in that: The color change information is a change value Y based on absorbance, an RGB value or a color change rule; wherein the change value Y based on absorbance is (I0-I) / I0, I0 is the absorbance at 521 nm when the reaction system lacks the target D3G, and I is the absorbance at 521 nm after the reaction system reacts with the target D3G.

10. A method for removing cryptic mycotoxin D3G by using the rapid identification method of cryptic mycotoxin D3G according to any one of claims 1 to 9, characterized in that: In step 1), the magnetic separation step removes D3G in the detection system.

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