Animal-derived food stimulant detection and analysis method

Abstract

The invention relates to the field of detection, in particular to an animal-derived food stimulant detection and analysis method, which comprises the following steps: detecting stimulants and metabolites thereof on samples of various parts of an animal body so as to determine the distribution of the stimulants and the metabolites thereof in the animal body; therefore, which parts in the animal body are only distributed with safety products can be determined, the corresponding positions on the generated animal body model are marked, and after the marked animal body model is output, a user can clearly and visually determine which parts of the animal body can be safely eaten. Therefore, the condition that people eat parts with relatively strong adverse effects on human bodies by mistake is avoided, and the body health of the people is ensured.

Description

Technical Field

[0001] The invention relates to the field of detection, and in particular to a method for detecting and analyzing stimulants in animal-derived food. Background Art

[0002] After being ingested by the human body, stimulants will be metabolized by the body and broken down into metabolites. The effects of metabolites on the human body are often weakened compared to the stimulants themselves, and as the degree of metabolism deepens, the effectiveness of the metabolites will disappear or become extremely weak so that they will not cause any effect on the human body.

[0003] Animals often secrete some endogenous stimulants, such as serotonin and glutamate. These stimulants and their metabolites are distributed differently in various parts of the animal body due to the action of the animal's circulatory system. In the animal body, if the metabolism degree of the stimulant in one part is low, then eating this part may have a greater impact on the human body. On the contrary, if the stimulant in one part is highly metabolized, then eating this part will often have less impact on the human body. However, the general public often cannot distinguish the distribution of stimulants and their metabolites in various parts of the animal body, and they are prone to physical discomfort due to eating these parts. Summary of the invention

[0004] Based on this, it is necessary to provide a method for detecting and analyzing stimulants in animal-derived foods to address the above-mentioned problems.

[0005] The embodiment of the present invention is implemented as follows: a method for detecting and analyzing stimulants in animal-derived food, the method comprising: S1: Select a stimulant from the stimulant database and generate a metabolic tree diagram of the stimulant, wherein the metabolic tree diagram includes several levels, the first level is the selected stimulant, and the following levels are the metabolites after the metabolic reaction of the stimulant, and the substances in each level are obtained by the metabolic reaction of the substances in the previous level; the metabolic conditions of the substance are marked at each substance except the substance in the lowest level in the metabolic tree diagram; and the corresponding action intensity is marked at each substance in the metabolic tree diagram; S2: taking samples from various parts of the animal body and pre-treating them respectively to obtain the test samples corresponding to each part, and then performing stimulant and metabolite detection on each test sample by liquid chromatography-tandem mass spectrometry to obtain the stimulant content and corresponding metabolite content of each part; S3: Generate an animal model, and mark the stimulant content and the corresponding metabolite content in each part of the animal model according to the test results; S4: Determine the food's circulation path in the human body to determine the metabolic environment at each point along the circulation path; S5: Obtain the action sites of stimulants and their corresponding metabolites in the human body; S6: For each metabolite with an action intensity above a preset intensity, determine whether the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity before flowing to the action site in the human body according to the metabolic environment at each location of the flow path and the metabolic conditions of the metabolite. If so, determine the metabolite as a safe product; otherwise, determine the metabolite as an unsafe product; S7: Each metabolite in the metabolic tree diagram with an action strength lower than the preset strength is also determined as a safe product; S8: Mark the areas where only safe products are distributed in the animal model, and output the animal model.

[0006] Preferably, the method of taking samples from various parts of the body and pre-treating them respectively to obtain the test samples corresponding to each part comprises: For each part of the animal body, weigh 5 g of the sample of that part into a centrifuge tube, add 100 μL of isotope internal standard working solution, add 10 mL of ammonium acetate buffer, vortex mix, adjust the pH to 10 with sodium hydroxide solution, add 15 mL of acetonitrile and vortex for 1 min, add salt bag for extraction, vortex for 1 min, let stand for 10 min, and centrifuge at 8000 r / min for 10 min; After centrifugation, add the solution to the EMR-lipids purification tube, add 3 mL of water, add 6 mL of acetonitrile extract, vortex for 1 min, centrifuge at 5000 r / min for 10 min, transfer all the supernatant to a 15 mL centrifuge tube, add the stripping salt bag, vortex for 1 min, centrifuge at 5000 r / min for 10 min, take 3 mL of acetonitrile layer and blow dry with nitrogen, add constant volume solution to constant volume, vortex through the membrane to obtain the test solution.

[0007] Preferably, the liquid chromatography-tandem mass spectrometer is connected to a computer device; the stimulant and metabolite detection is performed on each test sample by the liquid chromatography-tandem mass spectrometer, that is, the concentration of the stimulant in the test solution and the concentration of each metabolite of the stimulant are detected by the liquid chromatography-tandem mass spectrometer, and the detection results are transmitted to the computer device; The computer equipment calculates the content of any substance using the following formula: Where X is the content of the substance, is the concentration of the substance in the test solution, m is the sampling mass, and V is the fixed volume of the sample test solution.

[0008] Preferably, the chromatographic conditions include: Temperature: 45℃; Injection volume: 4 μL; Mobile phase A: methanol; mobile phase B: 0.1% formic acid; Flow rate: 0.3 mL / min; Mass spectrometry conditions included: Ion source: electrospray ion source, temperature 150°C; Scanning mode: multiple reaction monitoring; Collision gas: argon; Desolventization temperature: 500℃; Desolventizing gas flow rate: 1000L / h; Capillary voltage: 1.0 kV.

[0009] Preferably, step S1, and steps S3 to S8 are performed by a computer device; Select a stimulant from the stimulant database and generate a metabolic tree diagram of the stimulant including: S11: Get all metabolic reactions with the stimulant as a reactant; S12: Taking the stimulant as the first level, placing each metabolite of the obtained metabolic reaction at the next level of the stimulant, and connecting each metabolite to the stimulant through a branch line, wherein the metabolite connected to the stimulant is the metabolite of the nth level, and the initial value of n is 2; S13: for each metabolite in the nth layer, the metabolite is taken as a metabolite; S14: Obtain all metabolic reactions with metabolites as reactants; S15: placing the metabolites of each acquired metabolic reaction at the next level of the metabolized substance to obtain the metabolites of the n=n+1th level, and connecting the metabolites of this level with the metabolized substance through a branch line; S16: Repeat steps S13 to S15 until all metabolites at the lowest level can no longer be metabolized, thereby obtaining a metabolic dendrogram; S17: For each substance on the metabolic tree diagram, a metabolic condition is marked at the beginning of the branch line connecting the substance to the substance at the next level, wherein the metabolic condition includes the reaction temperature and the required enzyme; S18: Mark the intensity of the effect of each substance on the metabolic tree diagram on the human body, wherein the intensity of the effect is determined based on historical experimental data.

[0010] Preferably, determining the flow path of food in the human body to determine the metabolic environment at each point of the flow path includes: Generate a human perspective model; In the human perspective model, a digestive path is generated along the digestive tract from the mouth to the intestine; A blood path is generated along the blood circulation system starting from the intestine, wherein the direction of the blood path is consistent with the flow direction of the blood; A path formed by connecting the digestive path and the blood path is determined as a circulation path; Divide the flow path into several sections; For each section, the temperature and enzyme types of the section are determined based on the human body environment database to obtain the metabolic environment of the section, wherein the human body environment database includes the temperature and enzyme types of various parts of the human body.

[0011] Preferably, judging whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity before flowing to the action site in the human body based on the metabolic environment at each location of the flow path and the metabolic condition of the metabolite includes: Determine the site of action of the metabolite in a human perspective model; Determining a subpath from the oral cavity to the action location in the flow path; Determine in the metabolic tree diagram a metabolic pathway for the metabolite to be metabolized into a metabolite with an action intensity lower than a preset intensity, wherein the metabolic pathway starts from the metabolite and extends along a branch line toward a lower-level metabolite until it extends to a metabolite with an action intensity lower than the preset intensity; Whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity is determined based on the metabolic conditions of the substances passed by the metabolic pathway and the metabolic environment at each position of the subpathway.

[0012] Preferably, determining whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity based on the metabolic conditions of the substance passed by the metabolic pathway and the metabolic environment of each position of the subpathway includes: The corresponding metabolic conditions are sorted according to the order of substances passed through the metabolic pathway to obtain the first sequence; The metabolic environment of each position is sorted according to the order of the position in the subpathway to obtain a second sequence; Determining whether the second sequence includes a metabolic environment matching each metabolic condition in the first sequence, and if not, the metabolite cannot be metabolized into a metabolite with an action intensity lower than a preset intensity; If so, determine whether the order of the metabolic environment that matches the metabolic conditions in the first sequence is consistent with the order of the corresponding metabolic conditions in the first sequence. If not, the metabolite cannot be metabolized into a metabolite with an action intensity lower than the preset intensity. If so, the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity.

[0013] Preferably, the parts marked in the animal model where only safe products are distributed include: For each part in the animal model, determine whether the part contains only safe products; If so, render the part green; If not, the part is rendered in red, wherein the greater the maximum action intensity of the substance contained in the part is, the darker the rendered red is.

[0014] The present invention provides a method for detecting and analyzing stimulants in animal-derived food, the method comprising selecting a stimulant from a stimulant database and generating a metabolic dendrogram of the stimulant; taking samples from various parts of an animal body and pre-processing them respectively to obtain a detection sample corresponding to each part, and then performing stimulant and metabolite detection on each detection sample by a liquid chromatography-tandem mass spectrometer to obtain the stimulant content and the corresponding metabolite content of each part; generating an animal body model, and marking the stimulant content and the corresponding metabolite content of each part on the animal body model according to the detection results; determining the flow path of food in the human body to determine the metabolic environment at each location of the flow path; obtaining the action position of the stimulant and each corresponding metabolite in the human body; for each metabolite with an action intensity above a preset intensity, judging the location of the metabolite in the human body according to the metabolic environment at each location of the flow path and the metabolic condition of the metabolite. Whether it can be metabolized into a metabolite with an action intensity lower than a preset intensity before flowing to the action site in the human body, if so, the metabolite is determined as a safe product, otherwise, the metabolite is determined as an unsafe product; each metabolite with an action intensity lower than a preset intensity in the metabolic tree diagram is also determined as a safe product; the parts where only safe products are distributed are marked in the animal model, and the animal model is output; in the present application, stimulants and their metabolites are detected on samples of various parts of the animal body to determine the distribution of stimulants and their metabolites in the animal body, so that it is possible to determine which parts of the animal body have only safe products distributed and mark the corresponding positions on the generated animal model. After the marked animal model is output, the user can clearly and intuitively determine which parts of the animal body can be safely eaten, thereby avoiding accidental ingestion of parts that have strong adverse effects on the human body to ensure their health. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 A flowchart of a method for detecting and analyzing doping in animal-derived food provided in one embodiment; Figure 2 A diagram of the application environment of a method for detecting and analyzing doping in animal-derived food provided in one embodiment; Figure 3 A metabolic dendrogram of a method for detecting and analyzing doping in animal-derived food provided in one embodiment; Figure 4A schematic diagram of an animal model for a method for detecting and analyzing doping in animal-derived food provided in one embodiment; Figure 5 A schematic diagram of a human perspective model of a method for detecting and analyzing doping in animal-derived food provided in one embodiment; DETAILED DESCRIPTION In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0016] It is understood that the terms "first", "second", etc. used in the present invention may be used herein to describe various elements, but unless otherwise specified, these elements are not limited by these terms. These terms are only used to distinguish a first element from another element. For example, a first xx script may be referred to as a second xx script, and similarly, a second xx script may be referred to as a first xx script without departing from the scope of the present invention.

[0017] like Figure 1 As shown, in one embodiment, a method for detecting and analyzing stimulants in animal-derived food is proposed, the method comprising: S1: Select a stimulant from the stimulant database and generate a metabolic tree diagram of the stimulant, wherein the metabolic tree diagram includes several levels, the first level is the selected stimulant, and the following levels are the metabolites after the metabolic reaction of the stimulant, and the substances in each level are obtained by the metabolic reaction of the substances in the previous level; the metabolic conditions of the substance are marked at each substance except the substance in the lowest level in the metabolic tree diagram; and the corresponding action intensity is marked at each substance in the metabolic tree diagram; S2: taking samples from various parts of the animal body and pre-treating them respectively to obtain the test samples corresponding to each part, and then performing stimulant and metabolite detection on each test sample by liquid chromatography-tandem mass spectrometry to obtain the stimulant content and corresponding metabolite content of each part; S3: Generate an animal model, and mark the stimulant content and the corresponding metabolite content in each part of the animal model according to the test results; S4: Determine the food's circulation path in the human body to determine the metabolic environment at each point along the circulation path; S5: Obtain the action sites of stimulants and their corresponding metabolites in the human body; S6: For each metabolite with an action intensity above a preset intensity, determine whether the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity before flowing to the action site in the human body according to the metabolic environment at each location of the flow path and the metabolic conditions of the metabolite. If so, determine the metabolite as a safe product; otherwise, determine the metabolite as an unsafe product; S7: Each metabolite in the metabolic tree diagram with an action strength lower than the preset strength is also determined as a safe product; S8: Mark the areas where only safe products are distributed in the animal model, and output the animal model.

[0018] In this embodiment, if Figure 2 As shown, step S2 is performed by a staff member, step S1 and step S3 to step S8 are performed by a computer device, and the liquid chromatography-tandem liquid chromatography-tandem mass spectrometer in step S2 is connected to the computer device (such as Figure 2 As shown), the detection results can be transmitted to the computer device so that the computer device can perform subsequent data analysis based on the detection results; in addition, the computer device can be an independent physical server or terminal, or a server cluster composed of multiple physical servers, or a cloud server that provides basic cloud computing services such as cloud servers, cloud databases, cloud storage and CDN.

[0019] In this embodiment, the stimulant database is a preset database, which includes various endogenous stimulants in animals. Since the stimulants detected are endogenous stimulants in animals, animals of the same species all contain these stimulants, and the distribution of these stimulants and their metabolites in the animal body is basically the same as the detection result (because animals of the same species have the same metabolic mechanism and body circulation mechanism). Therefore, the distribution of each substance in the animal body model output by this embodiment can characterize the distribution of substances in all animals of this type; this method is executed for a stimulant, and the distribution of various substances obtained on the animal body model is only the distribution of this stimulant and the corresponding metabolites. If the distribution of other stimulants in the stimulant database on the animal body is determined, it is necessary to re-detect and analyze through this method; this method is executed for a kind of animal body, such as pigs, cattle, chickens, ducks, etc. If the material distribution of other animals is determined, it is also necessary to re-execute this method, that is, each execution of this method is for a stimulant and a kind of animal body.

[0020] In this embodiment, the selected stimulant is a stimulant that is harmful to the human body; the intensity of action, that is, the degree of harm of the stimulant to the human body, can be determined through preliminary experiments or from historical academic papers, and the experimental data in the academic papers are obtained based on the experimental results to pre-score the intensity of action of various stimulants and their metabolites, such as 1 point, 2 points, and 3 points. The higher the score, the greater the intensity of action.

[0021] In the present application, stimulants and their metabolites are detected on samples of various parts of the animal body to determine the distribution of stimulants and their metabolites in the animal body, so as to determine which parts of the animal body have only safe products and mark the corresponding positions on the generated animal body model. After the marked animal body model is output, the user can clearly and intuitively determine which parts of the animal body are safe to eat, thereby avoiding accidental ingestion of parts that have strong adverse effects on the human body to ensure their health.

[0022] As a preferred embodiment, the method of taking samples from various parts of the body and pre-processing them respectively to obtain the test samples corresponding to each part includes: For each part of the animal body, weigh 5 g of the sample of that part into a centrifuge tube, add 100 μL of isotope internal standard working solution, add 10 mL of ammonium acetate buffer, vortex mix, adjust the pH to 10 with sodium hydroxide solution, add 15 mL of acetonitrile and vortex for 1 min, add salt bag for extraction, vortex for 1 min, let stand for 10 min, and centrifuge at 8000 r / min for 10 min; After centrifugation, add the solution to the EMR-lipids purification tube, add 3 mL of water, add 6 mL of acetonitrile extract, vortex for 1 min, centrifuge at 5000 r / min for 10 min, transfer all the supernatant to a 15 mL centrifuge tube, add the stripping salt bag, vortex for 1 min, centrifuge at 5000 r / min for 10 min, take 3 mL of acetonitrile layer and blow dry with nitrogen, add constant volume solution to constant volume, vortex through the membrane to obtain the test solution.

[0023] The liquid chromatography-tandem mass spectrometer is connected to the computer device; the stimulant and metabolite detection is performed on each test sample by the liquid chromatography-tandem mass spectrometer, that is, the concentration of the stimulant in the test solution and the concentration of each metabolite of the stimulant are detected by the liquid chromatography-tandem mass spectrometer, and the detection results are transmitted to the computer device; The computer equipment calculates the content of any substance using the following formula: Where X is the content of the substance, is the concentration of the substance in the test solution, m is the sampling mass, and V is the fixed volume of the sample test solution.

[0024] Chromatographic conditions included: Temperature: 45℃; Injection volume: 4 μL; Mobile phase A: methanol; mobile phase B: 0.1% formic acid; Flow rate: 0.3 mL / min; Mass spectrometry conditions included: Ion source: electrospray ion source, temperature 150°C; Scanning mode: multiple reaction monitoring; Collision gas: argon; Desolventization temperature: 500℃; Desolventizing gas flow rate: 1000L / h; Capillary voltage: 1.0 kV.

[0025] In this embodiment, the unit of C is g / mL, the unit of V is mL, and the unit of m is g; in this embodiment, the part of the animal body is an edible part of the animal body, and a part can refer to an organ or a part of a set size on the animal body; in this embodiment, for each part, a test solution can be prepared for each substance (i.e., one of the stimulants and its metabolites), and then the test solution can be used to detect the content of the corresponding substance; different substances are detected, and the corresponding isotope internal standard working solutions are also configured differently. For example, if glutamate is to be detected, the isotope internal standard working solution of glutamate is pre-configured, and the configuration steps are as follows: accurately weigh 10 mg of glutamate standard (accurate to 0.00001 g), place it in a 10 mL brown volumetric flask, dissolve it with 0.1% formic acid methanol solution and dilute to the scale to prepare a standard stock solution with a concentration of 1000 μg / mL, and store it below -18°C; In this embodiment, a series of standard curve solutions (i.e., multiple solutions, each containing a specific concentration of a substance to be measured) are prepared before the determination, including multiple solutions of the substance to be measured with standard concentrations from small to large (such as 1 g / mL, 2 g / mL, 3 g / mL, etc.), and the solutions are measured in order from small to large concentrations, and the standard curve is drawn with the concentration of the substance to be measured as the horizontal coordinate and the peak area of ​​the isotope internal standard of the substance to be measured as the vertical coordinate; then the determination solution is measured to obtain a curve of the determination solution, and the position of the determination solution curve is compared with the standard curve, so as to quantify the concentration of the substance to be measured in the determination solution; the response value of the substance to be measured in the sample solution should be within the linear range of the standard curve. If the content of the substance to be measured is higher than the linear range of the standard curve, the standard curve should be re-prepared.

[0026] As a preferred embodiment, step S1, and steps S3 to S8 are performed by a computer device; like Figure 3 As shown, a stimulant is selected from the stimulant database, and a metabolic tree diagram of the stimulant is generated, including: S11: Get all metabolic reactions with the stimulant as a reactant; S12: Taking the stimulant as the first level, placing each metabolite of the obtained metabolic reaction at the next level of the stimulant, and connecting each metabolite to the stimulant through a branch line, wherein the metabolite connected to the stimulant is the metabolite of the nth level, and the initial value of n is 2; S13: for each metabolite in the nth layer, the metabolite is taken as a metabolite; S14: Obtain all metabolic reactions with metabolites as reactants; S15: placing the metabolites of each acquired metabolic reaction at the next level of the metabolized substance to obtain the metabolites of the n=n+1th level, and connecting the metabolites of this level with the metabolized substance through a branch line; S16: Repeat steps S13 to S15 until all metabolites at the lowest level can no longer be metabolized, thereby obtaining a metabolic dendrogram; S17: For each substance on the metabolic tree diagram, a metabolic condition is marked at the beginning of the branch line connecting the substance to the substance at the next level, wherein the metabolic condition includes the reaction temperature and the required enzyme; S18: Mark the intensity of the effect of each substance on the metabolic tree diagram on the human body, wherein the intensity of the effect is determined based on historical experimental data.

[0027] In this embodiment, all metabolic reactions with the stimulant as a reactant are obtained, that is, the reaction equations of all metabolic reactions with the stimulant as a reactant are obtained, including reactants, products, and reaction conditions (i.e., metabolic conditions). The reaction equations can be obtained from relevant academic materials on the Internet or determined through preliminary experiments, and are not limited here. Similarly, the reaction equations of all metabolic reactions with metabolites as reactants can also be obtained, and the corresponding metabolic conditions can be determined. In this embodiment, the generated metabolic dendrogram can clearly reflect the relationship between the stimulant and its metabolites and the reaction conditions, providing a basis for subsequent data analysis.

[0028] As a preferred embodiment, determining the flow path of food in the human body to determine the metabolic environment at each point of the flow path includes: Generate a human perspective model; In the human perspective model, a digestive path is generated along the digestive tract from the mouth to the intestine; A blood path is generated along the blood circulation system starting from the intestine, wherein the direction of the blood path is consistent with the flow direction of the blood; A path formed by connecting the digestive path and the blood path is determined as a circulation path; Divide the flow path into several sections; For each section, the temperature and enzyme types of the section are determined based on the human body environment database to obtain the metabolic environment of the section, wherein the human body environment database includes the temperature and enzyme types of various parts of the human body.

[0029] In this embodiment, the flow path is a directional line segment, that is, from the mouth to the intestine, and then from the intestine along the blood circulation system to the whole body; the computer device can use the pre-input human body structure (including internal structure and external structure) image as material, and then use these materials to generate a human body perspective model (such as Autodesk Maya) through pre-installed 3D modeling software (such as Autodesk Maya). Figure 5 Furthermore, animal models (such as Figure 4 The human body environment database is a preset database, and the data in it can be determined by users through relevant historical research data, which will not be elaborated here.

[0030] As a preferred embodiment, judging whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity before flowing to the action site in the human body based on the metabolic environment at each location of the flow path and the metabolic condition of the metabolite includes: Determine the site of action of the metabolite in a human perspective model; Determining a subpath from the oral cavity to the action location in the flow path; Determine in the metabolic tree diagram a metabolic pathway for the metabolite to be metabolized into a metabolite with an action intensity lower than a preset intensity, wherein the metabolic pathway starts from the metabolite and extends along a branch line toward a lower-level metabolite until it extends to a metabolite with an action intensity lower than the preset intensity; Whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity is determined based on the metabolic conditions of the substances passed by the metabolic pathway and the metabolic environment at each position of the subpathway.

[0031] Determining whether the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity based on the metabolic conditions of the substance passed by the metabolic pathway and the metabolic environment at each position of the subpathway includes: The corresponding metabolic conditions are sorted according to the order of substances passed through the metabolic pathway to obtain the first sequence; The metabolic environment of each position is sorted according to the order of the position in the subpathway to obtain a second sequence; Determining whether the second sequence includes a metabolic environment matching each metabolic condition in the first sequence, and if not, the metabolite cannot be metabolized into a metabolite with an action intensity lower than a preset intensity; If so, determine whether the order of the metabolic environment that matches the metabolic conditions in the first sequence is consistent with the order of the corresponding metabolic conditions in the first sequence. If not, the metabolite cannot be metabolized into a metabolite with an action intensity lower than the preset intensity. If so, the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity.

[0032] In this embodiment, the preset intensity may be an intensity of action with an intensity level of 3 points, which is not limited here; the metabolic conditions match the metabolic environment, that is, the enzymes and temperature in the metabolic environment are consistent with the enzymes and temperature in the metabolic conditions; different substances often act at different locations, and the circulation paths of each substance in the human body are not completely the same (mainly different blood circulation paths), so it is necessary to determine the subpath of the metabolite, that is, its specific circulation path in the human body, so as to determine the metabolic environment that the metabolite will pass through in the human body; in the second sequence, a metabolic environment matching each metabolic condition in the first sequence is determined, that is, indicating that there is a metabolic environment in the human body that can metabolize the metabolite into a metabolite with an action intensity lower than the preset intensity, but the metabolic tree diagram restrains metabolism in a sequence, and it is also necessary to determine whether the sequence of the metabolic environment in the human body is consistent with the sequence of the metabolic conditions. Only when the sequence of the metabolic environment is consistent with the sequence of the metabolic conditions can the human body metabolize the metabolite into a safe product.

[0033] As a preferred embodiment, the parts marked in the animal model where only safe products are distributed include: For each part in the animal model, determine whether the part contains only safe products; If so, render the part green; If not, the part is rendered in red, wherein the greater the maximum action intensity of the substance contained in the part is, the darker the rendered red is.

[0034] In this embodiment, the rendered animal body model can be output to a network platform, or directly output to the user terminal of each user; by rendering the animal body model in different colors, the audience who sees the animal body model can intuitively and clearly identify which parts are high-risk for consumption, and thus avoid these parts to ensure good health.

[0035] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0036] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for detecting and analyzing stimulants in animal-derived food, characterized in that: The method comprises: S1: Select a stimulant from the stimulant database and generate a metabolic tree diagram of the stimulant, wherein the metabolic tree diagram includes several levels, the first level is the selected stimulant, and the following levels are the metabolites after the metabolic reaction of the stimulant, and the substances in each level are obtained by the metabolic reaction of the substances in the previous level; the metabolic conditions of the substance are marked at each substance except the substance in the lowest level in the metabolic tree diagram; and the corresponding action intensity is marked at each substance in the metabolic tree diagram; S2: taking samples from various parts of the animal body and pre-treating them respectively to obtain the test samples corresponding to each part, and then performing stimulant and metabolite detection on each test sample by liquid chromatography-tandem mass spectrometry to obtain the stimulant content and corresponding metabolite content of each part; S3: Generate an animal model, and mark the stimulant content and the corresponding metabolite content in each part of the animal model according to the test results; S4: Determine the food's circulation path in the human body to determine the metabolic environment at each point along the circulation path; S5: Obtain the action sites of stimulants and their corresponding metabolites in the human body; S6: For each metabolite with an action intensity above a preset intensity, determine whether the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity before flowing to the action site in the human body according to the metabolic environment at each location of the flow path and the metabolic conditions of the metabolite. If so, determine the metabolite as a safe product; otherwise, determine the metabolite as an unsafe product; S7: Each metabolite in the metabolic tree diagram with an action strength lower than the preset strength is also determined as a safe product; S8: Mark the areas where only safe products are distributed in the animal model, and output the animal model.

2. The method according to claim 1, characterized in that The method of taking samples from various parts of the body and pre-treating them respectively to obtain the test samples corresponding to each part includes: For each part of the animal body, weigh 5 g of the sample of that part into a centrifuge tube, add 100 μL of isotope internal standard working solution, add 10 mL of ammonium acetate buffer, vortex mix, adjust the pH to 10 with sodium hydroxide solution, add 15 mL of acetonitrile and vortex for 1 min, add salt bag for extraction, vortex for 1 min, let stand for 10 min, and centrifuge at 8000 r / min for 10 min; After centrifugation, add the solution to the EMR-lipids purification tube, add 3 mL of water, add 6 mL of acetonitrile extract, vortex for 1 min, centrifuge at 5000 r / min for 10 min, transfer all the supernatant to a 15 mL centrifuge tube, add the stripping salt bag, vortex for 1 min, centrifuge at 5000 r / min for 10 min, take 3 mL of acetonitrile layer and blow dry with nitrogen, add constant volume solution to constant volume, vortex through the membrane to obtain the test solution.

3. The method according to claim 2, characterized in that The liquid chromatography-tandem mass spectrometer is connected to the computer device; the stimulant and metabolite detection is performed on each test sample by the liquid chromatography-tandem mass spectrometer, that is, the concentration of the stimulant in the test solution and the concentration of each metabolite of the stimulant are detected by the liquid chromatography-tandem mass spectrometer, and the detection results are transmitted to the computer device; The computer equipment calculates the content of any substance using the following formula: Where X is the content of the substance, is the concentration of the substance in the test solution, m is the sampling mass, and V is the fixed volume of the sample test solution.

4. The method according to claim 3, characterized in that Chromatographic conditions included: Temperature: 45℃; Injection volume: 4 μL; Mobile phase A: methanol; mobile phase B: 0.1% formic acid; Flow rate: 0.3 mL / min; Mass spectrometry conditions included: Ion source: electrospray ion source, temperature 150°C; Scanning mode: multiple reaction monitoring; Collision gas: argon; Desolventization temperature: 500℃; Desolventizing gas flow rate: 1000L / h; Capillary voltage: 1.0 kV.

5. The method according to claim 1, characterized in that Step S1, and steps S3 to S8 are performed by a computer device; Select a stimulant from the stimulant database and generate a metabolic tree diagram of the stimulant including: S11: Get all metabolic reactions with the stimulant as a reactant; S12: Taking the stimulant as the first level, placing each metabolite of the obtained metabolic reaction at the next level of the stimulant, and connecting each metabolite to the stimulant through a branch line, wherein the metabolite connected to the stimulant is the metabolite of the nth level, and the initial value of n is 2; S13: for each metabolite in the nth layer, the metabolite is taken as a metabolite; S14: Obtain all metabolic reactions with metabolites as reactants; S15: placing the metabolites of each acquired metabolic reaction at the next level of the metabolized substance to obtain the metabolites of the n=n+1th level, and connecting the metabolites of this level with the metabolized substance through a branch line; S16: Repeat steps S13 to S15 until all metabolites at the lowest level can no longer be metabolized, thereby obtaining a metabolic dendrogram; S17: For each substance on the metabolic tree diagram, a metabolic condition is marked at the beginning of the branch line connecting the substance to the substance at the next level, wherein the metabolic condition includes the reaction temperature and the required enzyme; S18: Mark the intensity of the effect of each substance on the metabolic tree diagram on the human body, wherein the intensity of the effect is determined based on historical experimental data.

6. The method according to claim 5, characterized in that Determine the food's flow path in the human body to determine the metabolic environment at each point in the flow path, including: Generate a human perspective model; In the human perspective model, a digestive path is generated along the digestive tract from the mouth to the intestine; A blood path is generated along the blood circulation system starting from the intestine, wherein the direction of the blood path is consistent with the flow direction of the blood; The path formed by connecting the digestive path and the blood path is determined as the circulation path; Divide the flow path into several sections; For each section, the temperature and enzyme types of the section are determined based on the human body environment database to obtain the metabolic environment of the section, wherein the human body environment database includes the temperature and enzyme types of various parts of the human body.

7. The method according to claim 6, characterized in that Judging whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity before flowing to the action site in the human body based on the metabolic environment at each location of the flow path and the metabolic conditions of the metabolite includes: Determine the site of action of the metabolite in a human perspective model; Determining a subpath from the oral cavity to the action location in the flow path; Determine in the metabolic tree diagram a metabolic pathway for the metabolite to be metabolized into a metabolite with an action intensity lower than a preset intensity, wherein the metabolic pathway starts from the metabolite and extends along a branch line toward a lower-level metabolite until it extends to a metabolite with an action intensity lower than the preset intensity; Whether the metabolite can be metabolized into a metabolite with an action intensity lower than a preset intensity is determined based on the metabolic conditions of the substances passed by the metabolic pathway and the metabolic environment at each position of the subpathway.

8. The method according to claim 7, characterized in that Determining whether the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity based on the metabolic conditions of the substance passed by the metabolic pathway and the metabolic environment at each position of the subpathway includes: The corresponding metabolic conditions are sorted according to the order of substances passed through the metabolic pathway to obtain the first sequence; The metabolic environment of each position is sorted according to the order of the position in the subpathway to obtain a second sequence; Determining whether the second sequence includes a metabolic environment matching each metabolic condition in the first sequence, and if not, the metabolite cannot be metabolized into a metabolite with an action intensity lower than a preset intensity; If so, determine whether the order of the metabolic environment that matches the metabolic conditions in the first sequence is consistent with the order of the corresponding metabolic conditions in the first sequence. If not, the metabolite cannot be metabolized into a metabolite with an action intensity lower than the preset intensity. If so, the metabolite can be metabolized into a metabolite with an action intensity lower than the preset intensity.

9. The method according to claim 7, characterized in that: The sites where only safe products were distributed in animal models include: For each part in the animal model, determine whether the part contains only safe products; If so, render the part green; If not, the part is rendered in red, wherein the greater the maximum action intensity of the substance contained in the part is, the darker the rendered red is.

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