Fluorescence detection method and kit for rapid detection of mycotoxin cross-linking phenol in dual-purpose raw materials

Abstract

The application discloses a fluorescence detection method and a kit for rapidly detecting mycotoxin roquefortine in a dual-purpose medicinal material, and a rapid fluorescence detection kit for mycotoxin roquefortine, which comprises the following components: A) a roquefortine standard solution; B) a roquefortine polyclonal antibody modified magnetic nano material; and C) a roquefortine polyclonal antibody modified up-conversion nano material. The detection method using the kit uses roquefortine polyclonal antibody modified magnetic nanoparticles to capture roquefortine and uses roquefortine polyclonal antibody modified up-conversion nano materials as a fluorescence probe to detect roquefortine, and has the advantages of simple method, high sensitivity and easy operation.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a fluorescence detection method and kit for rapid detection of the fungal toxin Alternaria alterniflora in medicinal and edible raw materials. Background Technology

[0002] Alternaria, a typical fungal toxin produced by Alternaria, possesses acute toxicity, genotoxicity, and potential carcinogenicity. It can cause symptoms of poisoning such as diarrhea and gastrointestinal bleeding, and is also associated with a high incidence of esophageal cancer in some regions, posing a serious threat to public health. This toxin easily contaminates food ingredients such as grains, oils, fruits, and vegetables, as well as medicinal and edible ingredients such as ginseng and wolfberry. Furthermore, the contamination is highly concealed, posing significant health risks through dietary intake.

[0003] Existing detection methods mainly rely on high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS), which suffer from drawbacks such as complex operation, long processing time, and dependence on large instruments, making them unsuitable for rapid on-site screening of medicinal and food raw materials. Fluorescence detection technology, on the other hand, offers advantages such as high sensitivity and ease of operation, making it suitable for rapid detection scenarios. Therefore, developing a rapid fluorescence detection method for lysozyme in both medicinal and food raw materials is of significant practical importance for ensuring food and drug safety and improving the toxin detection system. Summary of the Invention

[0004] The purpose of this invention is to provide a fluorescence detection method and kit for rapid detection of the mycotoxin Alternaria in food and medicinal materials, which is an immunofluorescence detection method for rapid and sensitive detection of Alternaria.

[0005] The present invention provides a fluorescence detection method for Alternaria alterniflora based on upconversion nanomaterials. It prepares polyclonal antibodies against Alternaria alterniflora and modifies them with magnetic nanoparticles and upconversion nanomaterials. The invented immunomagnetic particles and immunofluorescent materials can be applied to the rapid detection of Alternaria alterniflora.

[0006] A rapid fluorescent detection kit for the fungal toxin Alternaria alterniflora includes: A) Alternaria sol standard solution; B) Magnetic nanomaterials modified with a polyclonal antibody-linked cyclosporine; C) Upconversion nanomaterials modified with a polyclonal antibody-crosslinked spores.

[0007] The albedophenol standard solution uses albedophenol as a solute, and the concentration of albedophenol in the albedophenol standard solution is 0 mM and 0.008~0.012 mM, 0.04~0.06 mM, 0.08~0.12 mM, 0.45~0.55 mM.

[0008] The preparation of the magnetic nanomaterials modified with the polyclonal antibody-crosslinked spores specifically includes: Aminated Fe3O4 nanoparticles were dispersed in PBS buffer, sonicated, and then glutaraldehyde solution was added for the first reaction in a shaking incubator. After the reaction, the supernatant was removed by magnetic separation, the nanoparticles were collected, washed, and then dispersed in PBS buffer. After sonication, a polyclonal antibody of a concentration of cyclosporine was added, and the second reaction was carried out in a shaking incubator. After the reaction, the aptamer-functionalized magnetic nanoparticles were collected by magnetic separation.

[0009] The conditions for the first reaction were: 20~30 ℃, 60~100 rpm for 1~3 h; The conditions for the second reaction were: 35~40 ℃, 60~100 rpm for 8~12 h.

[0010] The preparation of the upconversion nanomaterial modified with the polyclonal antibody-crosslinked spores specifically includes: Upconversion nanomaterials were sonicated in PBS buffer, and then glutaraldehyde solution was added for the first shaking reaction. After the reaction, the mixture was centrifuged, the precipitate was washed, and the washed precipitate was dispersed in PBS buffer and sonicated. A solution of Alternaria polyclonal antibody was added, and after a second shaking reaction, the mixture was centrifuged and washed with PBS to obtain Alternaria polyclonal antibody-modified upconversion nanomaterials.

[0011] The conditions for the first shaking reaction were: shaking reaction at 20~30℃ for 1~4 h; The conditions for the second oscillation reaction were: 36-38℃, 50-150 rpm, for 10-14 h.

[0012] A fluorescent detection method for rapid detection of the mycotoxin albinosol in food and medicinal materials, using the aforementioned kit, specifically includes the following steps: S1: Mix the magnetic nanomaterial modified with polyclonal antibody of cyclosporine with the standard solution of cyclosporine. The mixture is shaken at 35~40℃ for the first reaction for 0.5~1.5h. After the reaction, the product is separated by magnetic field and washed. Then the mixture is resuspended in PBS buffer, and the upconversion nanomaterial modified with polyclonal antibody of cyclosporine is added for the second reaction. After the reaction, magnetic separation is performed and the fluorescence intensity is measured to prepare a standard curve. S2: Mix the magnetic nanomaterial modified with polyclonal antibody of co-electrosporin with the test solution. Shake the mixture at 35-40℃ for 0.5-1.5 h. After the reaction, separate the product with a magnetic field and wash it. Then resuspend the mixture in PBS buffer, add the upconversion nanomaterial modified with polyclonal antibody of co-electrosporin for a second reaction. After the reaction, perform magnetic separation and measure the fluorescence intensity. Quantify the co-electrosporin in the sample by combining the fluorescence intensity measured in the sample with the standard curve.

[0013] In step S1, the fluorescence intensity is measured under the conditions of excitation light of 980 nm and emission light of 542 nm. In step S2, the fluorescence intensity is measured under the conditions of excitation light at 980 nm and emission light at 542 nm.

[0014] Furthermore, the technical solution of the present invention is as follows: A. Preparation of polyclonal antibodies against Alternaria alternifolia; B. Preparation of aminoferric oxide magnetic nanoparticles; C. Modification of magnetic nanoparticles with polyclonal antibodies against Alternaria alternifolia; D. Preparation of upconversion nanomaterials; E. Modification of upconversion nanomaterials by polyclonal antibodies against Alternaria alternata.

[0015] F. Detection of Alternaria alterniflora using immunomagnetic nanoparticles and immunoupconversion nanomaterials.

[0016] The steps in step A for preparing the polyclonal antibody against Alternaria are as follows: A specific concentration of Alternaria alterniflora-BSA antigen was mixed with Freund's complete adjuvant 1 or Freund's incomplete adjuvant 1:1. The mixture was thoroughly stirred using a syringe to create a white, milky liquid that forms non-dispersible spheres when dropped into water. This liquid was used as the antigen. 0.2 mL of the antigen was injected subcutaneously into adult New Zealand white rabbits, with five injections per rabbit, for a total injection of 1.0 mL.

[0017] The preparation steps for aminoferric oxide magnetic nanoparticles in step B are as follows: Sodium acetate, 1,6-hexanediamine, and ferric chloride hexahydrate (FeCl3·6H2O) in a mass ratio of 2:6.5:1 were added to ethylene glycol at a volume of 3 times the total amount of reagents. After heating and stirring until homogeneous, the mixture was transferred to a reaction vessel and reacted at 200°C for several hours. After the reaction was completed and the reaction vessel cooled naturally, the supernatant was removed by magnetic separation using an external magnetic field. The black solid was collected, and the product was washed three times each with deionized water and ethanol. The product was then dried in a 60°C oven and collected to obtain aminated Fe3O4 nanoparticles.

[0018] The preparation steps of the magnetic nanoparticles modified with polyclonal antibody-crosslinked lysporin in step C are as follows: First, 2 mg / mL of aminated Fe3O4 nanoparticles were dispersed in 0.01 mol / L PBS (pH 7.4), and after sonication for 30 min, 25% glutaraldehyde solution (125 g / mL) was added. The nanoparticles were reacted with 100 mg / L Fe3O4 at 25 °C and 80 rpm for 2 h in a shaking incubator. After the reaction, the supernatant was removed by magnetic separation, the nanoparticles were collected, washed with 0.01 mol / L PBS, and then dispersed in 0.01 mol / L PBS solution. The mixture was sonicated for 5 min, and then 2 mg / mL of Alternaria polyclonal antibody (100 mg / L Fe3O4) was added. The mixture (L / mg Fe3O4) was incubated overnight at 37°C and 80 rpm in a shaking incubator. After the reaction, the aptamer-functionalized magnetic nanoparticles were collected by magnetic separation, washed three times with 0.01 mol / L PBS, dispersed in 1 mL of 0.01 mol / L PBS buffer, and stored at 4°C for later use.

[0019] The synthesis steps of the upconversion nanomaterials in step D are as follows: 1-Octadecene, oleic acid, yttrium chloride, ytterbium chloride, and erbium chloride in a molar ratio of 410:19:36:1:1 were maintained at 160°C for 30 minutes under argon protection. After cooling, methanol, sodium hydroxide, and ammonium fluoride in a molar ratio of 120:1.25:2 were added. The reaction was stirred for 30 minutes, then heated to 100°C under argon protection and maintained for one hour. The temperature was then increased to 300°C and maintained for 1 hour. After cooling to room temperature, the mixture was precipitated with anhydrous ethanol, washed with cyclohexane:anhydrous ethanol (2:1 ratio), and then centrifuged at 10,000 rpm for 10 minutes. Finally, it was dried in a vacuum drying oven at 60°C for 12 hours to obtain upconversion fluorescent nanomaterials. Then, 20 mg of the prepared UCNPs were added to 60 mL of isopropanol and sonicated for 30 min. 20 mL of distilled water and 2.5 mL of 25% ammonia were added, and the mixture was rapidly stirred magnetically for 15 min. Then, 20 mL of isopropanol and 60 L of TEOS mixture were added, and the reaction was carried out for 3 h. Subsequently, 30 mL of isopropanol and 200 L of APTES mixture were added, and the reaction was carried out for 1 h. After standing at room temperature for 2 h, the product was obtained by centrifugation. The product was washed several times with deionized water and then dried in a 60°C drying oven for 12 h to obtain SiO2-modified UCNPs with amino-modified surface.

[0020] The synthesis steps of the upconversion nanomaterial modified with cyclophosphamide polyclonal antibody in step E are as follows: A certain amount of upconversion nanoparticles were sonicated in PBS solution for 30 min, and then 25% glutaraldehyde solution (250 g / L) was added. L / 6 mg UCNPs), reacted with shaking at room temperature for 2 h, centrifuged at 12000 rpm for 10 min after 2 h, washed the precipitate three times with PBS buffer, then dispersed the precipitate in PBS buffer and sonicated for 10 min, then added 25 M chelazoline polyclonal antibody solution (40 The product (6 mg UCNPs) was reacted at 37°C with shaking at 100 rpm for 12 h, then centrifuged, washed three times with PBS, and dispersed in an appropriate amount of PBS to obtain a dispersion with a concentration of 6 mg / mL. The product was stored at 4°C. Step F, which utilizes immunomagnetic nanoparticles and immunoupconversion nanomaterials to detect Alternaria alterniflora, involves the following steps: 100 µL of magnetic nanoparticles modified with polyclonal antibody-coated intercalophenol were mixed with the sample to be tested, and the mixture was reacted with shaking at 37 °C for 1 h. After the reaction, the product was separated by a magnet and washed three times with PBS buffer, and then the mixture was resuspended in 400 µL of PBS buffer. Fluorescence intensity was measured under excitation light of 980 nm and emission light of 542 nm. A standard curve was prepared using a graded dilution of intercalophenol. The fluorescence intensity measured in the sample was used in conjunction with the standard curve to quantify intercalophenol in the sample.

[0021] Compared with the prior art, the present invention has the following advantages: This method combines the signal amplification effect of fluorescence upconversion nanomaterials to effectively improve detection sensitivity; at the same time, it utilizes magnetic nanoparticles modified with polyclonal antibodies against cyclosporine to effectively capture cyclosporine, simplifying sample pretreatment and extraction steps, and enabling fast detection speed. It can be used for on-site detection and screening with a handheld fluorescence instrument. Attached Figure Description

[0022] Figure 1 This is a transmission electron microscope image of an upconversion fluorescent nanomaterial.

[0023] Figure 2 The results show the detection results of different concentrations of albedophenol.

[0024] Figure 3 A standard curve for the detection of albedophenol using a rapid fluorescence detection method. Detailed Implementation

[0025] The technical solution of this application will be further described in detail below with reference to specific embodiments.

[0026] Example I. Preparation of Alternaria polyclonal antibodies (1) Preparation of antigen Dissolve 10 mg of Alternaria alternatasol (CAS No.: 641-38-3) in 200 μL of DMSO, and dissolve 6 mg of bovine serum albumin (BSA) in 6 mL of 0.1 M pH 4.5 acetate buffer. With magnetic stirring, add the Alternaria alternatasol solution dropwise to the BSA solution, then mix with 300 μL of 37% formaldehyde solution and react overnight at 30ºC. Centrifuge the reaction mixture, collect the supernatant, and dialyze against 1 L of pH 7.0 PBS at 4ºC for 2 days, changing the dialysate daily. Collect the dialysis antigen in a 1 mL centrifuge tube and store at -20ºC for later use.

[0027] (2) Antibody preparation Mix 1 mg / mL of Alternaria alterniflora-BSA antigen with Freund's complete adjuvant at a 1:1 ratio. Use a syringe to thoroughly mix the solution, creating a white, milky liquid that forms non-dispersible droplets in water. Inject 0.2 mL of this liquid subcutaneously into adult New Zealand white rabbits, 5 times per rabbit, for a total of 1.0 mL. For a second immunization, mix 1 mg / mL of Alternaria alterniflora-BSA antigen with Freund's incomplete adjuvant at a 1:1 ratio. Use a syringe to thoroughly mix the solution, creating a white, milky liquid that forms non-dispersible droplets in water. Inject 0.2 mL of this liquid subcutaneously into adult New Zealand white rabbits, 5 times per rabbit, for a total of 1.0 mL. Four weeks after the second immunization, administer a third immunization using the same antigen as the second immunization, 5 times per rabbit, for a total of 1.0 mL. Continue feeding the three-immunized New Zealand white rabbits for 4 weeks, then collect whole blood from their hearts for serum separation. The isolated serum was purified using the caprylic acid-ammonium sulfate method to remove impurities. The collected polyclonal immunoglobulins (polyclonal antibodies IgG) were dialyzed overnight with 0.01 mol / L PBS at pH 7.4 under light-protected conditions. The dialyzed polyclonal antibodies were collected and stored at -20 °C for later use.

[0028] II. Preparation of Magnetic Nanoparticles Modified with Polyclonal Antibody for Cross-linked Spore Powder for Rapid Separation of Cross-linked Spore Powder (1) Synthesis of amino magnetic nanoparticles 2.0 g of sodium acetate, 6.5 g of 1,6-hexanediamine, and 1.0 g of ferric chloride hexahydrate were added to 30 mL of ethylene glycol. After heating and stirring until homogeneous, the mixture was transferred to a 100 mL reactor and reacted at 200°C for several hours. After the reaction was completed and the reactor cooled naturally, the supernatant was removed by magnetic separation using an external magnetic field. The black solid was collected, and the product was washed three times each with deionized water and ethanol. The product was then dried in a 60°C oven and collected to obtain aminated Fe3O4 nanoparticles.

[0029] (2) Functional modification of amino magnetic nanoparticles using polyclonal antibodies against cross-linked spores 2 mg of aminoferric oxide was added to 1 mL of 0.01 mol / L pH 7.4 phosphate-buffered saline (PBS) and sonicated for 30 min. Then, 250 µL of 25% glutaraldehyde solution was added, and the reaction was carried out at 25℃ and 100 rpm for the first reaction. After 2 h of reaction, the supernatant was removed by magnetic separation. The reactants were washed with 1 mL of PBS buffer and then dispersed in 1 mL of PBS. After sonication for 5 min, 100 µL of 1 mg / mL Alternaria polyclonal antibody solution was added, and the reaction was carried out in a shaking incubator at 37℃ and 100 rpm for the second reaction for 10 h. After the reaction, the functionalized magnetic nanoparticles were collected by magnetic separation, washed three times with 1 mL of PBS buffer, and then dispersed in 1 mL of 0.01 mol / L pH 7.4 PBS and stored at 4℃.

[0030] III. Preparation of upconversion nanomaterials modified with polyclonal antibodies of cross-linked phenol for use as fluorescent probes (1) Synthesis and surface amino modification of upconversion nanoparticles Take 1 mmol of metal chloride (Y:Yb:Er=78:20:2), add oleic acid (OA) and 1-octadecene (ODE) to a 100 mL three-necked flask, stir and heat to 160 °C Hold at 30°C for 30 minutes, then cool to 50°C. C, add 10 mL of methanol solution (containing 2.5 mmol NaOH and 3.9 mmol NH4F), stir for 30 min, then heat to 100°C. The reaction was carried out at C for 1 hour, and finally heated to 300°C under argon protection. Keep at 60°C for 1 hour. After 1 hour, cool to room temperature, add ethanol to precipitate the product, wash and centrifuge the product three times, and then place it in a 60°C container. Dry in a drying oven at C for 12 hours.

[0031] Add 20 mg of UCNPs to 60 mL of isopropanol and sonicate for 30 min. Add 20 mL of distilled water and 2.5 mL of 25% ammonia, and stir rapidly with a magnetic stirrer for 15 min. Then add another 20 mL of isopropanol and 60 mL of distilled water. The L TEOS mixture was reacted for 3 h, followed by the addition of 30 mL isopropanol and 200 mL TEOS. The L APTES mixture was reacted for 1 h, then allowed to stand at room temperature for 2 h. After 2 h, the product was obtained by centrifugation, washed several times with deionized water, and then placed at 60°C. The SiO2-modified UCNPs with amino-modified surfaces were obtained by drying in a C drying oven for 12 h. TEM images of the prepared amino-modified iron oxide nanoparticles are shown below. Figure 1 As shown in the figure.

[0032] (2) Functional modification of upconversion nanomaterials using polyclonal antibodies against cyclosporine. Take 6 mg of amino-modified upconversion nanoparticles and sonicate them in 1 mL of PBS solution for 30 min, then add 250 mg of PBS solution. In a 25% glutaraldehyde solution, the mixture was reacted with shaking at 25°C for 2 hours. After 2 hours, it was centrifuged at 12000 rpm for 10 minutes. The precipitate was washed three times with PBS buffer, then dispersed in 1 mL of PBS buffer and sonicated for 10 minutes. 40 mL of PBS buffer was added. L 25 M-Alternaria polyclonal antibody solution was reacted at 37°C with shaking at 100 rpm for 12 h. After 12 h, the mixture was centrifuged, washed three times with PBS, and the product was dispersed in 1 mL PBS. After storage at C, upconversion nanoparticles modified with a polyclonal antibody were obtained.

[0033] IV. Detection of cross-linked polyclonal antibodies using magnetic nanoparticles and upconversion nanomaterials. First, put 100 L-Alternaria polyclonal antibody-modified magnetic nanoparticles and 100 The sample to be tested was mixed and the mixture was shaken at 37°C for 1 h. After the reaction, the product was separated by a magnet and washed with PBS buffer, and then 100 μL of the solution was added. L-Alternaria polyclonal antibody-modified upconversion nanomaterials were reacted with the mixture at 37°C for 1 h with shaking. The supernatant was then removed by magnetic separation, and the L-Alternaria polyclonal antibody-modified magnetic nanoparticle-L-Alternaria polyclonal antibody complex was washed three times with PBS buffer and then dispersed in 100 mL of PBS. In L PBS buffer, the fluorescence intensity of the dispersion at 542 nm was detected under 980 nm excitation light. Simultaneously, 100... L 0, 0.1, 0.2, 0.4, 0.8 After reacting the g / mL co-coated polyclonal antibody-modified magnetic nanoparticles with the co-coated polyclonal antibody according to the above steps, 100 g / mL co-coated polyclonal antibody-modified magnetic nanoparticles were added. The upconversion nanomaterials modified with L-alternaria polyclonal antibody were mixed and reacted with shaking at 37°C for 1 h. The products were separated by magnetism and washed three times with PBS buffer, and dispersed in 100 mL of PBS. In L PBS buffer, under 980 nm excitation light, the fluorescence intensity of the dispersion at 542 nm was measured to prepare a standard curve. The prepared standard curve is shown in Figure 1. Figure 2 As shown in Table 1, the detection limit of this method was calculated to be 0.62 mM. The concentration of albedophenol in the sample could be calculated using fluorescence intensity based on the standard curve. The recoveries of the samples were 111.01%, 102.78%, and 101.78% at artificial AOH contamination concentrations of 0.05, 0.10, and 0.20 mM, respectively. The results indicate that this method has good stability and accuracy.

[0034] Table 1. Detection of cross-linked phenols in food samples using a rapid immunofluorescence assay.

Claims

1. A rapid fluorescent detection kit for the fungal toxin Alternaria alterniflora, characterized in that, include: A) Alternaria sol standard solution; B) Magnetic nanomaterials modified with a polyclonal antibody-linked cyclosporine; C) Upconversion nanomaterials modified with a polyclonal antibody-crosslinked spores.

2. The rapid fluorescent detection kit for the fungal toxin Alternaria according to claim 1, characterized in that, The albedophenol standard solution uses albedophenol as a solute, and the concentration of albedophenol in the albedophenol standard solution is 0 mM and 0.008~0.012 mM, 0.04~0.06 mM, 0.08~0.12 mM, 0.45~0.55 mM.

3. The rapid fluorescent detection kit for the fungal toxin Alternaria according to claim 1, characterized in that, The preparation of the magnetic nanomaterials modified with the polyclonal antibody-crosslinked spores specifically includes: Aminated Fe3O4 nanoparticles were dispersed in PBS buffer, sonicated, and then glutaraldehyde solution was added for the first reaction in a shaking incubator. After the reaction, the supernatant was removed by magnetic separation, the nanoparticles were collected, washed, and then dispersed in PBS buffer. After sonication, a polyclonal antibody of a concentration of cyclosporine was added, and the second reaction was carried out in a shaking incubator. After the reaction, the aptamer-functionalized magnetic nanoparticles were collected by magnetic separation.

4. The rapid fluorescent detection kit for the fungal toxin Alternaria according to claim 3, characterized in that, The conditions for the first reaction were: 20~30 ℃, 60~100 rpm for 1~3 h; The conditions for the second reaction were: 35~40 ℃, 60~100 rpm for 8~12 h.

5. The rapid fluorescent detection kit for the fungal toxin Alternaria according to claim 1, characterized in that, The preparation of the upconversion nanomaterial modified with the polyclonal antibody-crosslinked spores specifically includes: Upconversion nanomaterials were sonicated in PBS buffer, and then glutaraldehyde solution was added for the first shaking reaction. After the reaction, the mixture was centrifuged, the precipitate was washed, and the washed precipitate was dispersed in PBS buffer and sonicated. A solution of Alternaria polyclonal antibody was added, and after a second shaking reaction, the mixture was centrifuged and washed with PBS to obtain Alternaria polyclonal antibody-modified upconversion nanomaterials.

6. The rapid fluorescent detection kit for the fungal toxin Alternaria according to claim 5, characterized in that, The conditions for the first shaking reaction were: shaking reaction at 20~30℃ for 1~4 h; The conditions for the second oscillation reaction were: 36-38℃, 50-150 rpm, for 10-14 h.

7. A fluorescent detection method for rapid detection of the mycotoxin albedoin in medicinal and edible raw materials, characterized in that, The kit described in any one of claims 1 to 6 specifically includes the following steps: S1: Mix the magnetic nanomaterial modified with polyclonal antibody of cyclosporine with the standard solution of cyclosporine. The mixture is shaken at 35~40℃ for the first reaction for 0.5~1.5h. After the reaction, the product is separated by magnetic field and washed. Then the mixture is resuspended in PBS buffer, and the upconversion nanomaterial modified with polyclonal antibody of cyclosporine is added for the second reaction. After the reaction, magnetic separation is performed and the fluorescence intensity is measured to prepare a standard curve. S2: Mix the magnetic nanomaterial modified with polyclonal antibody of co-electrosporin with the test solution. Shake the mixture at 35-40℃ for 0.5-1.5 h. After the reaction, separate the product with a magnetic field and wash it. Then resuspend the mixture in PBS buffer, add the upconversion nanomaterial modified with polyclonal antibody of co-electrosporin for a second reaction. After the reaction, perform magnetic separation and measure the fluorescence intensity. Quantify the co-electrosporin in the sample by combining the fluorescence intensity measured in the sample with the standard curve.

8. The fluorescence detection method for rapid detection of the mycotoxin albedoin in medicinal and edible raw materials according to claim 7, characterized in that, In step S1, the fluorescence intensity is measured under the conditions of excitation light of 980 nm and emission light of 542 nm. In step S2, the fluorescence intensity is measured under the conditions of excitation light at 980 nm and emission light at 542 nm.

9. The fluorescence detection method for rapid detection of the mycotoxin albedoin in medicinal and edible raw materials according to claim 7, characterized in that, In step S1, different concentrations of Alternaria alterniflora are detected and a standard curve is prepared.

10. The fluorescence detection method for rapid detection of the mycotoxin albedoin in medicinal and edible raw materials according to claim 7, characterized in that, The fluorescence intensity measured in step S2 is used to calculate the content of lysozyme in the test yangping using the standard curve.

Images