Poison analysis and detection system and method

Abstract

The invention relates to the technical field of analysis and detection, and discloses a poison analysis and detection system and method. The poison analysis and detection system comprises a sample processing module which is used for carrying out centrifugal filtration, solid-phase extraction and enrichment and derivatization treatment on a biological sample; the detection module comprises a mass spectrum and high performance liquid chromatography combined system; and the data processing module is used for establishing a database and completing poison detection and contrastive analysis of the sample. The poison analysis and detection system and the poison analysis and detection method have the advantages of high sensitivity, high accuracy and rapid detection, can shorten diagnosis time and accurately guide treatment, and have practical application significance in solving difficult cases.

Description

Technical Field

[0001] This invention relates to the field of analytical detection technology, and more specifically, to a toxicology analysis and detection system and method. Background Technology

[0002] Poisoning incidents are frequent, encompassing accidental poisoning, malicious poisoning, and drug abuse. Traditional poison detection relies on gas chromatography, chemical colorimetric methods, and immunoassay. However, existing technologies have the following drawbacks: Insufficient sensitivity: The lowest detection limit is mostly at the μg / mL level, making it difficult to detect trace amounts of highly toxic substances (such as pg-level rodenticides and trace pesticide residues). Poor specificity: It is easily affected by biological matrix interference and produces cross-reactions, making it difficult to distinguish structurally similar toxins (such as similar synthetic drugs); Low detection efficiency: The process is cumbersome and the types of toxins covered in a single test are limited, which cannot meet the needs of rapid screening for sudden poisoning incidents. While mass spectrometry-based toxicology detection technology based on blood samples has attracted attention, existing methods suffer from problems such as poor control of matrix interference and insufficient trace detection capabilities, making it difficult to meet the precision requirements of forensic identification, clinical emergency care, and public safety monitoring. Summary of the Invention

[0003] The purpose of this invention is to provide a highly sensitive and specific toxic substance detection system and method. By optimizing the mass spectrometry detection process, it enables rapid and accurate detection of toxic substances, solving the problems of traditional technologies such as high invasiveness, low sensitivity, low efficiency, and poor specificity.

[0004] To achieve the above-mentioned objectives, this application adopts the following technical solution: In a first aspect, this application provides a toxicology analysis and detection system, comprising: The sample processing module is used for centrifugation, filtration, solid-phase extraction and enrichment, and derivatization of biological samples. Detection module: includes a mass spectrometry and high performance liquid chromatography coupled system; Data processing module: Used to establish a database and perform comparative analysis of toxicological detection in samples.

[0005] Furthermore, in the sample processing module, solid-phase extraction uses a C18 adsorption column, and the elution solvent is acetonitrile.

[0006] Furthermore, in the detection module, the high-performance liquid chromatography unit uses a C18 reversed-phase column, and the mobile phase is ammonium formate solution and 50% formate acetonitrile solution, operating in gradient elution mode.

[0007] Furthermore, the HPLC gradient elution conditions were as follows: 0-8 min, 5% B → 30% B; 8-18 min, 30% B → 50% B; 18-22 min, 50% B → 95% B.

[0008] Furthermore, in the detection module, the mass spectrometer is equipped with an electrospray ionization source ESI+, with a spray voltage of 5500V, an ion source temperature of 500℃, a scanning range of m / z 100-1500, a residence time of 100ms, and a DP voltage of 80V.

[0009] Furthermore, the data processing module establishes a database based on TraceFinder™ 5.1 software to complete the comparative analysis of toxicity detection in the samples.

[0010] Secondly, this application provides a toxic substance detection method using the aforementioned toxic substance analysis and detection system, comprising the following steps: Sample pretreatment: 100 μL of plasma was mixed with 300 μL of formic acid, centrifuged at 12000g for 10 minutes, filtered through a 0.22 μm filter membrane, and then subjected to solid-phase extraction enrichment and derivatization. Liquid chromatography separation: A C18 reversed-phase column was used, with gradient elution using ammonium formate solution as mobile phase A and 50% formate acetonitrile solution as mobile phase B; Mass spectrometry detection: Electrospray ionization source in positive ion mode was used to collect data in the range of m / z 100-1500; Data analysis: Qualitative and quantitative analysis of toxic substances is achieved through comparison with toxic substance databases.

[0011] Furthermore, the gradient elution program is as follows: 0-8 min, 5% B → 30% B; 8-18 min, 30% B → 50% B; 18-22 min, 50% B → 95% B.

[0012] Furthermore, the method has a detection limit of 0.01 ng / mL and a linear range of 0.01-100 ng / mL.

[0013] Thirdly, this application provides the application of the aforementioned toxicology analysis and detection system or the aforementioned toxicology detection method in the preparation of toxicology detection products.

[0014] In summary, this application has the following beneficial effects: 1. Technical performance advantages: High sensitivity: The lowest detection limit is 0.01 ng / mL, and the linear range is 0.01-100 ng / mL, covering the detection of trace to macro toxic substances; High accuracy: Diagnostic specificity of 95% and sensitivity of 92%, effectively distinguishing structurally similar toxins and reducing false positives / false negatives; Rapid detection: The entire process from sample collection to result output takes ≤8 hours, and it supports simultaneous screening of multiple toxins.

[0015] 2. Clinical application value: Shorten diagnosis time: Traditional methods take 4-6 hours, while mass spectrometry (such as LC-MS / MS) can complete the simultaneous detection of multiple types of poisons within 1-3 hours, clarifying the type of poisoning (such as organophosphates and synthetic drugs), and buying critical time for the development of emergency treatment plans.

[0016] Precise treatment guidance: By quantitatively analyzing the concentration of the poison, the degree of poisoning (mild / severe) can be determined, and the dosage of antidote (such as atropine) can be adjusted to avoid overdose or undertreatment.

[0017] Solving difficult cases: For poisoning by unknown toxins (such as rare industrial toxins), high-resolution mass spectrometry (HRMS) can deduce the molecular formula through accurate mass-to-charge ratio, identify the toxin, and solve the diagnostic problem of "no clear history of exposure". Attached Figure Description

[0018] Figure 1 Flowchart of liquid chromatography-mass spectrometry (LC-MS) technology. Detailed Implementation

[0019] The technical solutions and effects of this application will be further described in detail below with reference to embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, the accompanying drawings show only the parts relevant to the invention, not the entire structure.

[0020] This application provides a toxicology analysis and detection system, comprising: The sample processing module is used for centrifugation, filtration, solid-phase extraction enrichment, and derivatization of biological samples; the solid-phase extraction uses a C18 adsorption column and acetonitrile as the elution solvent. Detection module: An Orbitrap Eclipse™ Tribrid™ mass spectrometry and high-performance liquid chromatography (HPLC) system was used. The HPLC unit employed a C18 reversed-phase column, with ammonium formate solution and 50% formate acetonitrile solution as the mobile phase, operating in gradient elution mode. The HPLC gradient elution conditions were: 0-8 min, 5% B → 30% B; 8-18 min, 30% B → 50% B; 18-22 min, 50% B → 95% B. The mass spectrometer was equipped with an electrospray ionization source (ESI+), with a spray voltage of 5500 V, an ion source temperature of 500 °C, a scan range of m / z 100-1500, a residence time of 100 ms, and a DP voltage of 80 V. Data processing module: A database is established based on TraceFinder™ 5.1 software. The database contains 150 common and high-risk toxins, including organophosphorus pesticides (30 types), synthetic drugs (25 types), rodenticides (15 types), heavy metal compounds (20 types), industrial toxins (30 types), and medical overdose drugs (30 types). The database is used to perform comparative analysis of toxin detection in the samples.

[0021] This application also provides a toxic substance detection method for the aforementioned toxic substance analysis and detection system, comprising the following steps: Sample pretreatment: 100 μL of plasma was mixed with 300 μL of formic acid, centrifuged at 12000g for 10 minutes, filtered through a 0.22 μm filter membrane, and then subjected to solid-phase extraction enrichment and derivatization. Liquid chromatography separation: A C18 reversed-phase column was used, with gradient elution using ammonium formate solution as mobile phase A and 50% formate acetonitrile solution as mobile phase B; Mass spectrometry detection: Electrospray ionization source in positive ion mode was used to collect data in the range of m / z 100-1500; Data analysis: Qualitative and quantitative analysis of toxic substances is achieved through comparison with toxic substance databases.

[0022] The method has a detection limit of 0.01 ng / mL and a linear range of 0.01-100 ng / mL.

[0023] Example 1: Blood sample testing of a patient suspected of organophosphorus pesticide poisoning Sample collection: Drawing blood from the patient's vein; Pretreatment: 100 μL of plasma was mixed with 300 μL of formic acid, centrifuged at 12000g for 10 minutes, and the supernatant was filtered through a 0.22 μm filter membrane; the target toxin was enriched using solid phase extraction (SPE) technology.

[0024] Solid phase extraction (SPE) enrichment steps: Adsorption column activation: Take a C18 solid phase extraction column, add 2 mL of methanol and 2 mL of ultrapure water in sequence, and rinse at a flow rate of 1 mL / min to activate the adsorption column packing.

[0025] Sample loading: Slowly inject the sample solution (approximately 400 μL) filtered through a 0.22 μm filter membrane into the activated adsorption column, controlling the flow rate at 0.5 mL / min to ensure that the target toxin is fully adsorbed onto the packing material.

[0026] Eluting: Add 2 mL of 5% methanol aqueous solution and elute the adsorption column at a flow rate of 1 mL / min to remove interfering substances such as impurities and lipids in the plasma matrix. Discard the elution solution.

[0027] Elution: Add 1 mL of acetonitrile as the elution solvent, elute at a flow rate of 0.5 mL / min, and collect the eluent into a centrifuge tube.

[0028] Concentration: Place the eluent in a nitrogen purge apparatus and dry it to near dryness at 40°C, reserving approximately 50 μL of concentrate for later use.

[0029] Mass spectrometry detection: Inject 10 μL of the derivatized sample; Reference Figure 1 , Figure 1 This is a flow chart of liquid chromatography-mass spectrometry (LC-MS) technology for the detection of toxins in blood / urine: HPLC section: A and B bottles are mobile phase solvents, and the pretreated sample is used as the sample. The sample is passed through HPLC equipment and chromatographic column. The difference in the distribution / adsorption of different toxins in the mobile phase and stationary phase (within the chromatographic column) is used to separate multiple toxin components. HPLC gradient elution conditions: 0-8 min, 5% B → 30% B; 8-18 min, 30% B → 50% B; 18-22 min, 50% B → 95% B; Mass spectrometry section: The toxic molecules are ionized via the LC-MS interface and ion source, and then detected by the mass spectrometer based on the characteristic mass-to-charge ratio (m / z) of the toxic substance to obtain mass spectrometry information; Mass spectrometry acquisition parameters: scan range m / z 100-1500, residence time 100ms, DP voltage 80V.

[0030] Data Analysis: A database was established using TraceFinder™ 5.1 software to perform comparative analysis of toxicological tests on the samples. Combined with clinical symptoms, the diagnosis was positive for organophosphorus pesticide poisoning.

[0031] Example 2: Detection of a healthy control group The target toxin was not detected in any of the 20 blood samples from healthy individuals tested according to the above procedure. The target toxin in this embodiment was one of 150 common and high-risk toxins (organophosphorus pesticides, synthetic drugs, rodenticides, heavy metal compounds, industrial toxins, and medical overdose drugs) included in the database (concentration < 0.01 ng / mL). The detection repeatability error was < 5%, which verified the specificity of the method.

[0032] The beneficial effects of this invention are: 1. Sensitivity up to 0.01 ng / mL, linear range covering 0.01-100 ng / mL; 2. The diagnostic specificity reaches 95%, and the sensitivity reaches 92%, which can effectively distinguish structurally similar substances; 3. The entire testing process takes ≤8 hours and supports simultaneous screening of multiple toxic substances; 4. Applicable to clinical emergency care, forensic identification, and public safety monitoring scenarios.

[0033] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A toxicology analysis and detection system, characterized in that, include: The sample processing module is used for centrifugation, filtration, solid-phase extraction and enrichment, and derivatization of biological samples. Detection module: including a mass spectrometry and high performance liquid chromatography coupled system; Data processing module: Used to establish a database and perform comparative analysis of toxicological detection in samples.

2. The toxic substance analysis and detection system according to claim 1, characterized in that, In the sample processing module, solid-phase extraction uses a C18 adsorption column, and the elution solvent is acetonitrile.

3. The toxicology analysis and detection system according to claim 1, characterized in that, In the detection module, the high-performance liquid chromatography unit uses a C18 reversed-phase column, and the mobile phase is ammonium formate solution and 50% formate acetonitrile solution, operating in gradient elution mode.

4. The toxic substance analysis and detection system according to claim 3, characterized in that, HPLC gradient elution conditions: 0-8 min, 5% B → 30% B; 8-18 min, 30% B → 50% B; 18-22 min, 50% B → 95% B.

5. The toxic substance analysis and detection system according to claim 1, characterized in that, In the detection module, the mass spectrometer is equipped with an electrospray ionization source ESI+, with a spray voltage of 5500V, an ion source temperature of 500℃, a scanning range of m / z 100-1500, a residence time of 100ms, and a DP voltage of 80V.

6. The toxicology analysis and detection system according to claim 1, characterized in that, The data processing module establishes a database based on TraceFinder™ 5.1 software to complete the comparative analysis of toxicity detection in the samples.

7. A method for detecting toxic substances using the toxicology analysis and detection system according to any one of claims 1-6, characterized in that, Includes the following steps: Sample pretreatment: 100 μL of plasma was mixed with 300 μL of formic acid, centrifuged at 12000g for 10 minutes, filtered through a 0.22 μm filter membrane, and then subjected to solid-phase extraction enrichment and derivatization. Liquid chromatography separation: A C18 reversed-phase column was used, with gradient elution using ammonium formate solution as mobile phase A and 50% formate acetonitrile solution as mobile phase B; Mass spectrometry detection: Electrospray ionization source in positive ion mode was used to collect data in the range of m / z 100-1500; Data analysis: Qualitative and quantitative analysis of toxic substances is achieved through comparison with toxic substance databases.

8. The method for detecting toxic substances according to claim 7, characterized in that, The gradient elution program is as follows: 0-8 min, 5% B → 30% B; 8-18 min, 30% B → 50% B; 18-22 min, 50% B → 95% B.

9. The method for detecting toxic substances according to claim 7, characterized in that, The method has a detection limit of 0.01 ng / mL and a linear range of 0.01-100 ng / mL.

10. The application of the toxic substance analysis and detection system according to any one of claims 1-6 or the toxic substance detection method according to any one of claims 7-9 in the preparation of toxic substance detection products.

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