Method for rapidly detecting content of aflatoxin B1 in traditional Chinese medicinal materials

Abstract

The application is suitable for the technical field of aflatoxin content detection, and particularly relates to a method for rapidly detecting the content of aflatoxin B1 in traditional Chinese medicinal materials, comprising the following steps: step S1: weighing a preset weight of traditional Chinese medicinal material samples, crushing the samples, screening the samples through a particle size controlled screen to obtain uniform powder samples, and then putting the powder samples into centrifugal tubes; step S2: adding an appropriate amount of pure water into the centrifugal tubes, uniformly dispersing the water through ultrasonic dispersion to obtain a mixed solution, and adjusting the pH value of the mixed solution to a specific range by using an acid-base regulator to ensure that the subsequent adsorption effect is optimal. The application significantly improves the extraction efficiency and detection sensitivity of aflatoxin B1 in samples by introducing magnetic nanomaterials and ultrasonic auxiliary technology. By adjusting the pH value of the mixed solution to a specific range, the adsorption effect of aflatoxin B1 in the solution is optimal, and the use amount of solvent is reduced, and environmental pollution is reduced.

Description

Technical Field

[0001] This invention relates to the field of aflatoxin content detection technology, specifically a method for rapidly detecting aflatoxin B1 content in traditional Chinese medicine. Background Technology

[0002] Aflatoxin B1 is a highly toxic fungal metabolite commonly found in natural plant samples such as grains, oilseeds, and traditional Chinese medicinal herbs. It exhibits strong carcinogenic, mutagenic, and teratogenic effects. Detecting and controlling the aflatoxin B1 content in traditional Chinese medicinal herbs is a crucial step in ensuring their quality and safety. Existing methods for aflatoxin B1 detection mainly include high-performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), and solid-phase extraction-gas chromatography (SPE-GC). These methods have a certain degree of sensitivity and accuracy in practical applications and have become commonly used methods for detecting aflatoxin B1 in traditional Chinese medicinal herbs and food.

[0003] However, traditional HPLC detection methods typically require complex sample pretreatment, such as solvent extraction, liquid-liquid partitioning, and multiple centrifugations, making sample processing cumbersome and consuming large amounts of organic solvents, which can easily cause environmental pollution. Furthermore, impurities in the sample can interfere with the detection of aflatoxin B1, affecting the accuracy of the assay. While ELISA is relatively simple to operate, its results are often subject to error due to limitations in reagent stability and specificity, and it is difficult to adapt to the detection requirements of complex matrix samples. Although SPE-GC technology offers high sensitivity, it has a long detection time, high equipment costs, and potential sample loss during operation. Summary of the Invention

[0004] This invention provides the following technical solution:

[0005] A rapid method for detecting aflatoxin B1 content in traditional Chinese medicinal materials includes the following steps:

[0006] Step S1: Weigh the preset weight of Chinese medicinal material sample, crush it, and sieve it through a sieve with particle size control to obtain a uniform powder sample. Then put the powder sample into a centrifuge tube.

[0007] Step S2: Add an appropriate amount of pure water to the centrifuge tube and disperse it evenly by ultrasonication to obtain a mixed solution. Use an acid-base adjuster to adjust the pH value of the mixed solution to a specific range to ensure the best subsequent adsorption effect.

[0008] Step S3: Add the pre-weighed magnetic nanomaterial particles to the mixed solution with the adjusted pH value, and mix thoroughly to ensure that the magnetic nanomaterials are uniformly dispersed in the solution;

[0009] Step S4: The added mixed solution is subjected to ultrasonic extraction, and an external magnetic field is used to cause the magnetic nanomaterial particles to rapidly separate into layers under the action of the external magnetic field. After separation, the supernatant is collected and filtered with filter paper to ensure that impurities are minimized in subsequent processing.

[0010] Step S5: Add anhydrous methanol to the precipitate bound to the magnetic nanomaterial particles to desorb aflatoxin B1. After sonication for 5 minutes, the mixture is separated again under an external magnetic field. The supernatant is collected, and a small amount of the desorbed liquid is taken. Aflatoxin B1 is determined by ultra-high performance liquid chromatography (UHPLC) to calculate the content of aflatoxin B1 in the sample.

[0011] Step S6 involves performing the operations of steps S1-S5 on multiple samples of Chinese medicinal materials. The samples are processed in batches using multi-channel centrifugation and a magnetic rack. At the same time, the instrument is calibrated using aflatoxin B1 standard solution, and a standard curve is plotted to ensure the accuracy and stability of the determination.

[0012] Furthermore, in step S2, the ultrasound power is 200W and the duration is 10 minutes.

[0013] Furthermore, in step S3, the magnetic nanomaterial used is Fe3O4 nanoparticles with a particle size of 50-100 nm, and the amount added is 0.1 g per gram of sample.

[0014] Furthermore, adjust the pH of the mixed solution to 6.0-7.0.

[0015] Furthermore, in step S4, the magnetic field strength of the external magnetic field is 5000 Gauss, and the magnetic nanomaterials and solution are rapidly separated through the magnetic force frame, reducing the sample separation time.

[0016] Furthermore, in step S4, the supernatant after stratification is further filtered through filter paper to remove residual suspended particles and improve the purity of the test sample.

[0017] Furthermore, in step S5, anhydrous methanol is used as the desorption solvent, and the desorption process is carried out for 5 minutes at an ultrasonic power of 200W.

[0018] Furthermore, in step S6, the ultra-high performance liquid chromatograph is calibrated using aflatoxin B1 standard solution before each batch of samples is tested, and a standard curve is plotted to ensure the accuracy of the test results.

[0019] Furthermore, in step S6, automated data analysis software is used to process the sample test data, automatically generate a test report, and conduct a compliance assessment of the aflatoxin B1 content of the sample according to the standard limit.

[0020] The present invention provides a rapid method for detecting aflatoxin B1 content in traditional Chinese medicinal materials, which has the following beneficial effects:

[0021] By introducing magnetic nanomaterials and ultrasound-assisted technology, the extraction efficiency and detection sensitivity of aflatoxin B1 in samples were significantly improved. Adjusting the pH of the mixed solution to a specific range optimized the adsorption of aflatoxin B1 in the solution, while reducing solvent usage and environmental pollution. Magnetic separation technology under an external magnetic field shortened sample separation time, avoiding the cumbersome steps of multiple centrifugations, and improving detection speed and sample recovery rate. Furthermore, the use of anhydrous methanol desorption combined with ultra-high performance liquid chromatography (UHPLC) effectively quantifies the content of aflatoxin B1, ensuring the accuracy of the results. This method also exhibits good repeatability, is suitable for batch processing of multiple samples, and can significantly improve detection efficiency. Compared with existing technologies, this invention has advantages such as simple operation, rapid detection, high sensitivity, and environmental friendliness, and is particularly suitable for rapid screening and quality control of aflatoxin B1 in traditional Chinese medicine materials, showing broad application prospects. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0023] The specific implementation of the present invention will be described in detail below with reference to specific embodiments.

[0024] This invention provides a method for rapidly detecting aflatoxin B1 content in traditional Chinese medicinal materials, comprising the following steps:

[0025] Step S1: Weigh the preset weight of Chinese medicinal material sample, crush it, and sieve it through a sieve with particle size control to obtain a uniform powder sample. Then put the powder sample into a centrifuge tube.

[0026] Step S2: Add an appropriate amount of pure water to the centrifuge tube and disperse it evenly by ultrasonication to obtain a mixed solution. Use an acid-base adjuster to adjust the pH value of the mixed solution to a specific range to ensure the best subsequent adsorption effect.

[0027] Step S3: Add the pre-weighed magnetic nanomaterial particles to the mixed solution with the adjusted pH value, and mix thoroughly to ensure that the magnetic nanomaterials are uniformly dispersed in the solution;

[0028] Step S4: The added mixed solution is subjected to ultrasonic extraction, and an external magnetic field is used to cause the magnetic nanomaterial particles to rapidly separate into layers under the action of the external magnetic field. After separation, the supernatant is collected and filtered with filter paper to ensure that impurities are minimized in subsequent processing.

[0029] Step S5: Add anhydrous methanol to the precipitate bound to the magnetic nanomaterial particles to desorb aflatoxin B1. After sonication for 5 minutes, the mixture is separated again under an external magnetic field. The supernatant is collected, and a small amount of the desorbed liquid is taken. Aflatoxin B1 is determined by ultra-high performance liquid chromatography (UHPLC) to calculate the content of aflatoxin B1 in the sample.

[0030] Step S6 involves performing the operations of steps S1-S5 on multiple samples of Chinese medicinal materials. The samples are processed in batches using multi-channel centrifugation and a magnetic rack. At the same time, the instrument is calibrated using aflatoxin B1 standard solution, and a standard curve is plotted to ensure the accuracy and stability of the determination.

[0031] In one embodiment of the present invention, by sieving the pulverized Chinese medicinal material sample using a particle size-controlled sieve, a powder sample with uniform particle size can be obtained. This processing method ensures the homogeneity of the sample and avoids differences in extraction efficiency caused by uneven particle size, thereby improving the stability and repeatability of the detection results. Accurately weighing the pretreated sample and placing it into a centrifuge tube helps ensure consistent sample quality for each test and reduces detection errors.

[0032] Adding an appropriate amount of pure water to the sample and then dispersing it thoroughly using ultrasound allows for better contact between the active ingredients and the solvent, thereby increasing the dissolution rate of aflatoxin B1. Adjusting the pH of the mixed solution to a specific range enhances the effect of aflatoxin B1 in subsequent adsorption steps, making it more easily adsorbed by magnetic nanomaterials under specific pH conditions. Compared with traditional solvent extraction methods, this step significantly improves the extraction efficiency of aflatoxin B1 and reduces the use of organic solvents.

[0033] After adding magnetic nanomaterial particles, a vortex mixer is used for thorough mixing to ensure uniform dispersion of the magnetic nanomaterials in the solution, thereby improving the adsorption efficiency of aflatoxin B1. The magnetic nanomaterials can rapidly bind to aflatoxin B1 in the solution, and their surface activity makes the adsorption process more efficient and rapid, exhibiting a larger specific surface area and faster adsorption rate compared to traditional adsorption materials.

[0034] During ultrasonic extraction, an external magnetic field is used to rapidly separate the magnetic nanomaterials into layers. Using a magnetic rack, the separation process can be completed in a short time, and the supernatant is collected. The supernatant is then further filtered with filter paper to remove suspended impurities, ensuring the purity of the sample before desorption. This step is more efficient than traditional centrifugation methods, reducing separation time and minimizing sample loss that may occur due to multiple centrifugations.

[0035] Anhydrous methanol was added to magnetic nanomaterials containing aflatoxin B1 for desorption. The aflatoxin B1 was then fully desorbed by ultrasonic treatment, followed by a second separation using an external magnetic field. The supernatant was then subjected to ultra-high performance liquid chromatography (UHPLC) to determine the aflatoxin B1 content. Compared to traditional detection methods, this approach not only improves the desorption rate of the target analyte but also shortens the desorption time, making the detection process much faster.

[0036] With the support of multi-channel centrifugation and a magnetic rack, multiple samples can be processed simultaneously, enabling batch processing and improving detection efficiency. Aflatoxin B1 standard solution is used for instrument calibration and standard curve plotting to ensure the accuracy and stability of each measurement. Data is processed by automated analysis software, which not only reduces human error but also quickly generates test reports, improving overall detection efficiency. Compared with existing technologies, this method has significant advantages in sample processing speed, detection accuracy, and environmental friendliness, making it particularly suitable for large-scale rapid detection of traditional Chinese medicinal materials.

[0037] In this embodiment, in step S2, the ultrasonic power is 200W and the duration is 10 minutes. This ensures that the Chinese medicinal material sample is fully dispersed in a short time, increases the contact area between the active ingredients and the solvent, thereby enhancing the dissolution efficiency of aflatoxin B1, ensuring the homogeneity of the sample, and thus improving the accuracy of detection.

[0038] In this embodiment, the magnetic nanomaterial used in step S3 is Fe3O4 nanoparticles with a particle size of 50-100 nm, and the amount added is 0.1 g per gram of sample. Fe3O4 nanoparticles have a large specific surface area and excellent magnetic responsiveness, which can rapidly and efficiently adsorb aflatoxin B1. The particle size of 50-100 nm can provide high adsorption capacity, while ensuring particle dispersibility and solution stability, effectively improving adsorption efficiency and detection sensitivity.

[0039] In this embodiment, the pH of the mixed solution was adjusted to 6.0-7.0. Within this pH range, aflatoxin B1 is chemically stable in solution, which is more conducive to its binding with magnetic nanomaterials. At the same time, it avoids solubility changes caused by excessively high or low pH, ensuring the best adsorption process and thus improving the accuracy of detection.

[0040] In this embodiment, in step S4, the applied magnetic field strength is 5000 Gauss. The magnetic field, acting as a magnetic frame, enables rapid stratification of the magnetic nanomaterials and the solution, reducing sample separation time. The high-intensity magnetic field can rapidly aggregate magnetic nanoparticles, achieving rapid separation of the magnetic material from the liquid phase in the sample. This not only shortens the separation time but also reduces sample processing steps, avoiding sample degradation or errors caused by prolonged operation and improving overall detection efficiency.

[0041] In this embodiment, in step S4, the supernatant after separation is further filtered through filter paper to remove residual suspended particles and improve the purity of the sample. This process effectively removes any remaining impurities or suspended matter, ensuring sample cleanliness in subsequent ultra-high performance liquid chromatography analysis, preventing column blockage or interference peaks, thereby ensuring the stability of the determination and the accuracy of the results.

[0042] In this embodiment, in step S5, anhydrous methanol is used as the desorption solvent, and the desorption process is carried out for 5 minutes at an ultrasonic power of 200W. Anhydrous methanol has strong dissolving power and can efficiently desorb aflatoxin B1 adsorbed by magnetic nanomaterials. Combined with ultrasonic treatment, the desorption process can be accelerated, ensuring complete release of aflatoxin B1, thereby improving desorption efficiency, reducing desorption time, and optimizing the detection process.

[0043] In this embodiment, in step S6, the ultra-high performance liquid chromatograph is calibrated using aflatoxin B1 standard solution before each batch of samples is tested, and a standard curve is plotted to ensure the accuracy of the measurement results. Calibration with standard solution and plotting a standard curve allow for adjustments to the instrument's sensitivity and accuracy, ensuring the reliability of each measurement, effectively reducing errors caused by instrument drift, and ensuring the consistency and accuracy of test results for different batches of samples.

[0044] In this embodiment, in step S6, automated data analysis software is used to process the sample test data, automatically generate a test report, and assess the compliance of the aflatoxin B1 content of the sample according to the standard limit. Using automated data analysis software can quickly process large amounts of test data, avoiding errors and workload from manual calculations, ensuring the accuracy and speed of data processing. Simultaneously, it can automatically determine whether the sample is qualified according to the standard limit, simplifying the result analysis process and improving testing efficiency and report generation speed.

[0045] The following is an experimental report based on the above technical solution, covering the experimental background, objectives, materials and methods, results and analysis, and conclusions. The tables summarize the different steps and experimental results.

[0046] Experimental Report: A Rapid Method for Detecting Aflatoxin B1 Content in Traditional Chinese Medicines

[0047] Experimental Background: This experiment aims to verify a rapid method for detecting aflatoxin B1 content in traditional Chinese medicine based on magnetic nanomaterial adsorption technology. By optimizing the sample pretreatment, adsorption, desorption, and detection processes, the accuracy, efficiency, and environmental friendliness of the detection can be improved.

[0048] Experimental objective:

[0049] Optimize the extraction and detection process of aflatoxin B1 in traditional Chinese medicinal materials.

[0050] To determine the effectiveness and optimal parameters of magnetic nanomaterials in adsorbing aflatoxin B1.

[0051] Improve the batch processing capacity of testing and the accuracy of results.

[0052] Materials and Methods:

[0053]

[0054] Experimental Results and Analysis:

[0055] Note:

[0056] R of the standard curve 2 The value is 0.998, indicating that the detection method has a good linear relationship and is suitable for quantitative analysis of aflatoxin B1.

[0057] The relative standard deviation (RSD) was less than 3%, indicating that the experimental method had good repeatability.

[0058] According to national standards, aflatoxin B1 content exceeding 5.0 μg / kg is considered unqualified. Samples with excessive levels of B1 and B2 were found to be compliant, while the remaining samples were compliant.

[0059] in conclusion:

[0060] This experiment achieved efficient adsorption and separation of aflatoxin B1 from traditional Chinese medicine using Fe3O4 magnetic nanomaterials. Combined with an external magnetic field, sample separation was rapid, and ultrasound-assisted desorption further improved detection efficiency.

[0061] Adjusting the pH of the mixed solution to the range of 6.0-7.0 helps to improve the adsorption effect of aflatoxin B1, ensuring the accuracy and sensitivity of the detection.

[0062] The use of ultra-high performance liquid chromatography (UHPLC) for detection, combined with standard solution calibration and automated data analysis, makes the detection results more reliable and suitable for rapid screening of batch samples.

[0063] Compared with traditional methods, this method has significant advantages in sample processing speed, detection accuracy, environmental friendliness, and ease of operation. It is particularly suitable for rapid screening and quality control of large batches of Chinese medicinal materials and has broad application prospects.

[0064] Discussion and suggestions:

[0065] It is recommended that the amount of magnetic nanomaterials added be further optimized in practical applications to meet the detection needs of different types of Chinese medicinal material samples.

[0066] For samples with high content, it is recommended to dilute them and retest to ensure the accuracy of the test results.

[0067] Developing higher-throughput sample processing equipment could be considered to further improve detection efficiency.

[0068] This experimental report provides strong technical support for the rapid detection of aflatoxin B1 in traditional Chinese medicine through detailed descriptions of experimental procedures, analysis of data results, and a summary of the method's advantages. The tables clearly display the parameters and results of each step, facilitating experimental reproduction and subsequent optimization.

[0069] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials, characterized in that, Includes the following steps: Step S1: Weigh the preset weight of Chinese medicinal material sample, crush it, and sieve it through a sieve with particle size control to obtain a uniform powder sample. Then put the powder sample into a centrifuge tube. Step S2: Add an appropriate amount of pure water to the centrifuge tube and disperse it evenly by ultrasonication to obtain a mixed solution. Use an acid-base adjuster to adjust the pH value of the mixed solution to a specific range to ensure the best subsequent adsorption effect. Step S3: Add the pre-weighed magnetic nanomaterial particles to the mixed solution with the adjusted pH value, and mix thoroughly to ensure that the magnetic nanomaterials are uniformly dispersed in the solution; Step S4: The added mixed solution is subjected to ultrasonic extraction, and an external magnetic field is used to cause the magnetic nanomaterial particles to rapidly separate into layers under the action of the external magnetic field. After separation, the supernatant is collected and filtered with filter paper to ensure that impurities are minimized in subsequent processing. Step S5: Add anhydrous methanol to the precipitate bound to the magnetic nanomaterial particles to desorb aflatoxin B1. After sonication for 5 minutes, the mixture is separated again under an external magnetic field. The supernatant is collected, and a small amount of the desorbed liquid is taken. Aflatoxin B1 is determined by ultra-high performance liquid chromatography (UHPLC) to calculate the content of aflatoxin B1 in the sample. Step S6 involves performing the operations of steps S1-S5 on multiple samples of Chinese medicinal materials. The samples are processed in batches using multi-channel centrifugation and a magnetic rack. At the same time, the instrument is calibrated using aflatoxin B1 standard solution, and a standard curve is plotted to ensure the accuracy and stability of the determination.

2. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S2, the ultrasonic power is 200W and the duration is 10 minutes.

3. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S3, the magnetic nanomaterial used is Fe3O4 nanoparticles with a particle size of 50-100 nm, and the amount added is 0.1 g per gram of sample.

4. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S3, the pH of the mixed solution is adjusted to 6.0-7.

0.

5. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S4, the magnetic field strength of the applied magnetic field is 5000 Gauss. The magnetic nanomaterials and the solution are rapidly separated through the action of the magnetic frame, reducing the sample separation time.

6. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S4, the supernatant after separation is further filtered through filter paper to remove residual suspended particles and improve the purity of the test sample.

7. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S5, anhydrous methanol is used as the desorption solvent, and the desorption process is carried out for 5 minutes at an ultrasonic power of 200W.

8. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S6, the ultra-high performance liquid chromatograph is calibrated using aflatoxin B1 standard solution before each batch of samples is tested, and a standard curve is plotted to ensure the accuracy of the test results.

9. The method for rapid detection of aflatoxin B1 content in traditional Chinese medicinal materials according to claim 1, characterized in that, In step S6, automated data analysis software is used to process the sample test data, automatically generate a test report, and conduct a compliance assessment of the aflatoxin B1 content of the sample according to the standard limit.

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