CN-121994985-A - Multi-dimensional high-precision detection method and system for biotoxin in agricultural products

CN121994985ACN 121994985 ACN121994985 ACN 121994985ACN-121994985-A

Abstract

The invention discloses a multi-dimensional high-precision detection method and a multi-dimensional high-precision detection system for biotoxin in agricultural products, which comprise the steps of carrying out aseptic collection and grading pretreatment on agricultural product samples to obtain uniform samples, extracting the uniform samples by adopting a mixed solvent system, adding internal standard substances, salting out and layering, collecting supernatant, sequentially carrying out QuEChERS purification, multi-mechanism adsorption column purification and molecular imprinting polymer specific enrichment on the supernatant, carrying out dual-mode detection on target toxin enrichment liquid, respectively carrying out quantitative detection by adopting a liquid chromatography-tandem mass spectrometry, carrying out screening detection by adopting an intelligent electrochemical sensing method, carrying out optimization treatment on dual-mode detection signals, and comprehensively judging sample detection results.

Inventors

QU WEIFENG
ZHANG DA
XU CHAOYING
WANG PEI
ZHU JUNJIAN
CHEN MENGXIA

Assignees

浙江商技检测有限公司

Dates

Publication Date: 20260508
Application Date: 20260204

Claims (10)

1. A multi-dimensional high-precision detection method for biotoxin in agricultural products is characterized by comprising the following steps: carrying out aseptic collection and grading pretreatment on an agricultural product sample to obtain a homogenized sample; extracting the homogenized sample by adopting a mixed solvent system, adding an internal standard substance, salting out and layering, and collecting supernatant; sequentially performing QuEChERS purification, multiple mechanism adsorption column purification and molecular imprinting polymer specific enrichment on the supernatant to obtain target toxin enrichment liquid; Performing dual-mode detection on the target toxin enrichment liquid, quantitatively detecting by adopting a liquid chromatography-tandem mass spectrometry method, and screening and detecting by adopting an intelligent electrochemical sensing method; And processing the dual-mode detection signal by adopting a machine learning algorithm, and comprehensively judging the detection result of the multiclass biotoxin in the sample by combining with a preset standard limit value.
2. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: The hierarchical preprocessing further comprises: Removing impurities from a solid agricultural product sample, and grinding and screening; degreasing a high-fat agricultural product sample; homogenizing a high-moisture agricultural product sample; filtering and impurity-removing treatment is carried out on the liquid agricultural product sample; And (5) placing the collected sample in a light-proof sterile container for low-temperature storage and transportation.
3. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: the mixed solvent system comprises a mixed system containing nitriles, water and organic acid; In the extraction process of extracting the homogenized sample by adopting a mixed solvent system, vortex oscillation and ultrasonic extraction are combined; in the salting-out layering process, a combined system of anhydrous magnesium sulfate and sodium chloride is adopted, and supernatant fluid is collected after centrifugal treatment.
4. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: the QuEChERS purification adopts an adsorbent combination system; The adsorbent combination system comprises an amine adsorbent and a carbon-based adsorbent; And after the QuEChERS is purified, collecting supernatant through centrifugal treatment, and entering the multi-mechanism adsorption column purification step.
5. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: The multi-mechanism adsorption column adopts a purification column based on the synergistic effect of hydrophobic effect, ion exchange and hydrogen bonding; before the multiple mechanism adsorption column is purified, activating the multiple mechanism adsorption column, controlling the flow rate of liquid to finish purification, and collecting effluent liquid; the molecularly imprinted polymer is formed by electropolymerization on the surface of a magnetic nanocomposite substrate by taking target biotoxin as a template molecule; And in the specific enrichment process of the molecularly imprinted polymer, the steps of oscillation adsorption, magnetic field separation, washing and elution are carried out, the eluent in the elution step is subjected to nitrogen blow-drying treatment, a methanol-water mixed system is adopted as constant volume liquid, and after dissolution, the eluent is filtered by a filter membrane to obtain the liquid to be detected.
6. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: the chromatographic conditions of the liquid chromatography-tandem mass spectrometry include: the chromatographic column is a C 18 reversed phase chromatographic column; column temperature 40 ℃; the sample injection amount is 5 mu L; Mobile phase A is 0.1% formic acid water solution, mobile phase B is methanol; gradient elution procedure: 0-2min,30% B, 2-10min,30 80% B, 10-15min, 80; from 80% to 80% B, 10%; 15min, 80; the flow rate was 0.3mL/min.
7. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: mass spectrometry conditions of the liquid chromatography-tandem mass spectrometry include: the ion source is an ESI source and is ionized in a positive ion mode; Spraying voltage is 3.5kV; the ion source temperature is 350 ℃; the air curtain pressure is 20psi; The atomization air pressure is 50psi; Drying air pressure 60psi; A multiple reaction monitoring mode was employed.
8. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: The machine learning algorithm comprises a baseline correction algorithm and a characteristic region screening algorithm; The method comprises the steps of processing a dual-mode detection signal by adopting a machine learning algorithm, and further comprises the steps of adopting the baseline correction algorithm to correct baseline drift of an electrochemical detection signal, and adopting the characteristic region screening algorithm to optimize the quantitative relation between the detection signal and the toxin concentration; The comprehensive judgment of the detection results of the multiple biotoxins in the sample further comprises the step of judging the results based on the consistency comparison of the double-mode detection results.
9. The method for multi-dimensional, high-precision detection of biotoxins in agricultural products according to claim 1, characterized in that: The multi-class biotoxin comprises aflatoxins, fusarium toxins, ochratoxins and other common agricultural product pollution biotoxins; The agricultural products comprise grains, oil crops, fruits and vegetables and silage.
10. A biotoxin multi-dimensional high-precision detection system in agricultural products based on the biotoxin multi-dimensional high-precision detection method in agricultural products as set forth in any one of claims 1-9, characterized in that the system includes: the collection module is used for carrying out aseptic collection and grading pretreatment on the agricultural product sample to obtain a uniform sample; The pretreatment module is used for extracting the homogenized sample by adopting a mixed solvent system, adding an internal standard substance, salting out and layering, and collecting supernatant; The purification and enrichment treatment module is used for sequentially carrying out QuEChERS purification, multi-mechanism adsorption column purification and molecular imprinting polymer specific enrichment on the supernatant to obtain target toxin enrichment liquid; the dual-mode detection module is used for carrying out dual-mode detection on the target toxin enrichment liquid, respectively carrying out quantitative detection by adopting a liquid chromatography-tandem mass spectrometry method, and carrying out screening detection by adopting an intelligent electrochemical sensing method; and the result re-judging module is used for processing the dual-mode detection signal by adopting a machine learning algorithm and comprehensively judging the detection result of the multi-class biotoxin in the sample by combining with a preset standard limit value.

Description

Multi-dimensional high-precision detection method and system for biotoxin in agricultural products Technical Field The invention relates to the technical field of detection, in particular to a method and a system for detecting biotoxin in agricultural products in a multi-dimensional and high-precision manner. Background In the whole chain of planting, harvesting, storing and processing agricultural products, the agricultural products are easy to be infected by fungi, bacteria and other microorganisms to generate biotoxin, wherein the mycotoxin (such as aflatoxin, zearalenone, ochratoxin A and the like) seriously threatens the food safety and public health due to the characteristics of wide distribution, strong toxicity, easy residue and the like. However, the traditional detection method is mostly aimed at single toxin, has the defects of complicated pretreatment, long detection period, strong matrix interference, insufficient synchronous detection capability of multiple toxins and the like, and is difficult to meet the detection requirements of modern agriculture on high flux, high precision and quick response. Disclosure of Invention The invention aims to solve the problems in the prior art, and provides a method and a system for multi-dimensional and high-precision detection of biotoxin in agricultural products, which further solve all or one of the problems in the prior art. In order to solve the technical problems, the specific technical scheme of the invention is as follows: In one aspect, the invention provides a method for multi-dimensional high-precision detection of biotoxin in agricultural products, comprising the following steps: carrying out aseptic collection and grading pretreatment on an agricultural product sample to obtain a homogenized sample; extracting the homogenized sample by adopting a mixed solvent system, adding an internal standard substance, salting out and layering, and collecting supernatant; sequentially performing QuEChERS purification, multiple mechanism adsorption column purification and molecular imprinting polymer specific enrichment on the supernatant to obtain target toxin enrichment liquid; Performing dual-mode detection on the target toxin enrichment liquid, quantitatively detecting by adopting a liquid chromatography-tandem mass spectrometry method, and screening and detecting by adopting an intelligent electrochemical sensing method; And processing the dual-mode detection signal by adopting a machine learning algorithm, and comprehensively judging the detection result of the multiclass biotoxin in the sample by combining with a preset standard limit value. Further, the hierarchical preprocessing further includes: Removing impurities from a solid agricultural product sample, and grinding and screening; degreasing a high-fat agricultural product sample; homogenizing a high-moisture agricultural product sample; filtering and impurity-removing treatment is carried out on the liquid agricultural product sample; And (5) placing the collected sample in a light-proof sterile container for low-temperature storage and transportation. Further, the mixed solvent system comprises a mixed system containing nitriles, water and organic acid; In the extraction process of extracting the homogenized sample by adopting a mixed solvent system, vortex oscillation and ultrasonic extraction are combined; in the salting-out layering process, a combined system of anhydrous magnesium sulfate and sodium chloride is adopted, and supernatant fluid is collected after centrifugal treatment. Further, the QuEChERS purification adopts an adsorbent combination system; The adsorbent combination system comprises an amine adsorbent and a carbon-based adsorbent; And after the QuEChERS is purified, collecting supernatant through centrifugal treatment, and entering the multi-mechanism adsorption column purification step. Further, the multi-mechanism adsorption column adopts a purification column based on the synergistic effect of hydrophobic effect, ion exchange and hydrogen bonding; before the multiple mechanism adsorption column is purified, activating the multiple mechanism adsorption column, controlling the flow rate of liquid to finish purification, and collecting effluent liquid; the molecularly imprinted polymer is formed by electropolymerization on the surface of a magnetic nanocomposite substrate by taking target biotoxin as a template molecule; And in the specific enrichment process of the molecularly imprinted polymer, the steps of oscillation adsorption, magnetic field separation, washing and elution are carried out, the eluent in the elution step is subjected to nitrogen blow-drying treatment, a methanol-water mixed system is adopted as constant volume liquid, and after dissolution, the eluent is filtered by a filter membrane to obtain the liquid to be detected. Further, the chromatographic conditions of the liquid chromatography-tandem mass spectrometry include: the chromatographic column is a