A method for detecting an acute poisoning drug in a biological sample
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- DALIAN INSTITUTE OF CHEMICAL PHYSICS CHINESE ACADEMY OF SCIENCES
- Filing Date
- 2024-11-27
- Publication Date
- 2026-05-29
AI Technical Summary
Existing rapid on-site detection methods for acute poisoning have poor sensitivity, poor selectivity, and are prone to false alarms and false negatives.
An ion trap mass spectrometer equipped with an electrospray ionization source was used, combined with liquid-liquid extraction, heating pre-desorption, high-selectivity ionization, and dual confirmation of parent and fragment ions, to establish a toxic substance characteristic spectrum library. By comparing the detection results with the spectra in the database, toxic substances can be identified with high sensitivity and high selectivity.
It achieves highly sensitive, selective, and accurate detection of acute poisoning drugs, improving the reliability of rapid on-site detection.
Smart Images
Figure CN122109264A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of medical diagnostics, specifically relating to a rapid on-site detection method for acute poisoning with high sensitivity and high identification accuracy. Background Technology
[0002] Rapid and accurate detection of blood poisoning plays a crucial role in medical emergency care, forensic medicine, and environmental monitoring. Poisoning incidents often require swift response, and timely and accurate detection of blood poisoning components not only provides clinicians with vital information for treatment but also offers key evidence for forensic investigations. However, the complexity of blood poisoning detection cannot be ignored, as the concentrations are typically low and the complex blood matrix can interfere with test results. With the rapid development of the chemical industry, the types of chemical toxins are increasing, and their structures are becoming more complex, making the need for rapid screening, qualitative, and quantitative detection increasingly urgent. Therefore, developing blood poisoning detection methods and technologies with high sensitivity, high accuracy, and rapid detection capabilities is of significant practical importance.
[0003] Currently, the detection techniques for blood poisons are mainly divided into traditional chemical analysis methods and modern instrumental analysis methods. Traditional methods such as chemical titration and colorimetric analysis have been gradually replaced by instrumental analysis methods such as high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-mass spectrometry (LC-MS). Among them, GC-MS and LC-MS combine the separation capabilities of chromatography with the high sensitivity and high resolution of mass spectrometry, enabling precise qualitative and quantitative analysis of blood poison components. However, the sample pretreatment process for LC-MS is complex and the detection cycle is long, making it unsuitable for medical personnel; moreover, LC-MS instruments are expensive, making it difficult to meet the needs of rapid detection in clinical settings or on-site. In recent years, the development of portable and miniaturized detection technologies has provided new directions for poison detection. For example, portable instruments based on immunoassay, ion mobility spectrometry (IMS), Raman spectroscopy, and sensors have received widespread attention due to their rapid response and real-time detection capabilities. However, these methods cannot simultaneously meet the requirements of universality, identification accuracy, and sensitivity, thus limiting their application. Mass spectrometry is widely recognized as the "gold standard" for qualitative analysis. In addition to retaining the advantages of laboratory mass spectrometry, such as good versatility and wide applicability, miniature mass spectrometry also meets the requirements of portability and fast analysis speed for field applications. Moreover, its hardware cost is significantly reduced compared to laboratory mass spectrometry.
[0004] Wang Mingcheng et al. invented a rapid on-site testing kit for acute poisoning (patent number CN96204993.X), which includes a base and a lid. The base has six partitions parallel to the short side of the testing kit, creating equidistant divisions. Another compartment has three horizontal partitions parallel to the long side of the testing kit. The remaining five compartments each hold 13 testing boxes for 56 kinds of toxic compounds in 8 categories, including pesticides, sleeping pills, rodenticides, alkaloids, inorganic toxins, strong acids, low-carbon fatty alcohols, and toxic gases. However, the kit suffers from drawbacks such as low sensitivity, poor selectivity, and susceptibility to false alarms.
[0005] Liu Rui et al. invented a rapid testing device for acute poisoning on-site emergency treatment (patent number CN202311184978.3), which relates to the field of emergency testing kit technology. The device includes a base with a drive box fixedly connected to the top. It comprises a base plate, side plates, and a top plate. Rotating a transmission column causes the drive shaft to rotate via a second pulley. The drive shaft, through a second bevel gear, drives a first threaded rod, causing a slider to move the base plate and side plates. The operator rotates a worm gear, which, through a worm wheel, drives the threaded column to rotate. The moving block moves the top plate to an appropriate height, opens the top cover, and removes the rapid testing instrument from the placement chamber. The instrument is placed on the top plate platform on the right side of the device for testing. Test tubes used in the testing can be placed inside a placement hole on the left side of the device for rapid testing of toxic substances, facilitating emergency treatment of patients. The top plate facilitates the testing of toxic substances. However, this device involves multiple chemical reagents and suffers from low sensitivity and a tendency to false alarms.
[0006] To address the problems of poor sensitivity, poor selectivity, and susceptibility to false alarms and false negatives in existing rapid on-site detection methods for acute poisoning, this invention provides a method for detecting acute poisoning drugs in biological samples based on micro ion trap mass spectrometry. This method combines liquid-liquid extraction, pre-desorption by heating, highly selective ionization, and dual confirmation of parent and fragment ions to achieve highly sensitive, selective, and accurate identification of acute poisoning drugs, providing important support for the treatment of acute poisoning. Summary of the Invention
[0007] The technical problem to be solved by this invention is that existing rapid on-site detection methods for acute poisoning suffer from poor sensitivity, poor selectivity, and are prone to false alarms and false negatives. The invention aims to develop a rapid on-site detection method based on ion trap mass spectrometry.
[0008] The specific content includes:
[0009] An ion detection instrument equipped with an electrospray ion source is used; the ion detection instrument is an ion trap mass spectrometer.
[0010] 200-1000 μL (preferably 400-600 μL) of biological sample is dropped into the groove of the sample sheet. Then, a methanol-acetonitrile mixture with a volume ratio of 1:10 to 1:1 (preferably 1:9) is used as an elution solution to rinse the sample droplets. Finally, the rinsing solution is ionized by an electrospray ionization source and then enters the detection instrument for detection.
[0011] The grooved sample sheet refers to a sheet-like sample sheet with one or more grooves on its surface;
[0012] The eluent is used to rinse the sample at a flow rate of 1 nL / min to 1 ml / min (preferably 10 uL / min) and a volume of 0.5 uL to 1 ml (preferably 5 uL).
[0013] The results of the above tests were compared with the toxic substance characteristic spectrum library in the testing instrument to determine the type of poison in the sample.
[0014] 1) Establishment of a database of characteristic spectra of common toxic substances:
[0015] a. The establishment of the toxic substance standard database employs an ion detection instrument equipped with an electrospray ionization source. Common toxic substance standards are diluted into three or more standard solutions of 0.1-1 μg / mL using a methanol-acetonitrile mixture. These standard solutions are then subjected to spray ionization within the electrospray ionization source. Ion detectors are then used to detect the toxic substances in both precursor and daughter ion modes. The spectra and peak positions of the precursor and daughter ions for different toxic substances are recorded and entered into the database as a basis for determining the presence of toxic substances.
[0016] If an ion trap mass spectrometer is used as a detector, fragment ion information can be obtained by adjusting the working mode of the ion trap mass analyzer to the collision dissociation mode based on the information of the parent ion, and then entered into the database as a basis for judging whether there are toxic substances.
[0017] b. Establishment of a simulated sample database containing toxic substances: Take a sample and add one of the common toxic substance standards with a final concentration of 0.1-1 μg / mL to it to obtain a target sample solution; use a syringe to draw 200-1000 μL of the target sample solution and place it on a grooved sample plate; use a methanol-acetonitrile mixture to flush the target solution into the capillary inlet, and then perform spray ionization and detection by an ion detector;
[0018] Finally, a spectral library of toxic substances was established, including the characteristic ion peak information of the parent ion and daughter ion corresponding to each toxic substance;
[0019] 2) Sample Analysis: Using a syringe, 200-1000 μL of target sample solution is drawn and placed on a grooved sample plate. The target solution is then flushed with a methanol-acetonitrile mixture to allow it to enter the capillary inlet, followed by spray ionization and detection. The sample is compared with the sample database containing toxic substances from step 1) to check for a corresponding parent ion peak for any toxic substance recorded in the toxic substance characteristic spectrum library. If no corresponding parent ion peak is found, the blood sample does not contain any toxic substance recorded in the toxic substance characteristic spectrum library. If a corresponding parent ion peak is found in the database, that parent ion is screened and subjected to collision dissociation mode detection to obtain its daughter ion information. The daughter ions are then compared: if a daughter ion matches one in the database, toxic substance information is provided. If no toxic substance corresponding to a parent ion in the database is detected, the detection is complete, and a message indicating that no toxic substance information was found in the database is provided.
[0020] The poisons include one or more of the following four categories: pesticides, sedatives and hypnotics, therapeutic drugs, and narcotics.
[0021] Specifically, it includes one or more of the following: quizalofop-P-ethyl, carbofuran, dimethoate, amitraz, omethoate, risperidone, chlorpromazine, clozapine, clonazepam, diazepam, acetaminophen, difenidol, chlorpheniramine, methamphetamine, methadone, noscapine, ketamine, amphetamine, AM-2201, and JWH-018.
[0022] The sample sheet is one or more of aluminum foil, copper foil, and gold foil.
[0023] The depth of the groove is 0.1µm to 5µm (preferably 0.5µm), and the spacing between adjacent grooves is 1µm to 1mm (preferably 0.5mm).
[0024] Biological samples are one or more of the following: saliva, urine, or blood from a person or animal.
[0025] Ion trap mass spectrometry is either discontinuous injection ion trap mass spectrometry or continuous injection ion trap mass spectrometry equipped with an electrospray ion source. Attached Figure Description
[0026] Figure 1 A schematic diagram of a method for detecting acute poisoning drugs in biological samples based on micro ion trap mass spectrometry; wherein: Figure 1 (a) Omethoate; (b) Carbofuran; (c) Quizalofop-P-ethyl; (d) Risperidone; (e) Diazepam; (f) Chlorpromazine; (g) Acetaminophen; (h) Chlorpheniramine and (i) Difenidol, all at a concentration of 0.1 ng / μl;
[0027] Figure 2A spectral analysis of acute poisoning drugs in biological samples based on micro-ion trap mass spectrometry; 1 ng / μl of omethoate (a)U in blood. floatDC When it is -10V and (b)U floatDC This is the mass spectrum at -30V. Detailed Implementation
[0028] An ion detection instrument equipped with an electrospray ion source was used; the ion detection instrument was an ion trap mass spectrometer.
[0029] A 200 μL biological sample is dropped into the groove of the sample slide, and then the sample droplet is rinsed with a 1:9 volume ratio elution buffer. Finally, the rinsing liquid is ionized by an electrospray ionization source and then enters the detection instrument for detection.
[0030] The grooved sample sheet refers to a sheet-like sample sheet with two parallel strip-shaped grooves on its surface;
[0031] The sample is placed in the groove, and the eluent is flushed by flowing through the sample along the length of the groove at a flow rate of 10 uL / min and a volume of 5 uL.
[0032] The results of the above tests were compared with the toxic substance characteristic spectrum library in the testing instrument to determine the type of poison in the sample.
[0033] Establishment of a database of characteristic spectra of common toxic substances:
[0034] a. The establishment of the toxic substance standard database employed an ion detection instrument equipped with an electrospray ionization source. Common toxic substance standards were diluted to concentrations of 0.1, 0.3, 0.5, 0.8, 0.9, and 1 μg / mL using a 1:9 methanol-acetonitrile mixture. These standard solutions were then subjected to spray ionization within the electrospray ionization source, and detected separately using an ion-type detector in both precursor and daughter ion modes. The spectra and peak positions of the precursor and daughter ions for different toxic substances were recorded and entered into the database as a basis for determining the presence of toxic substances.
[0035] If an ion trap mass spectrometer is used as a detector, fragment ion information can be obtained by adjusting the working mode of the ion trap mass analyzer to the collision dissociation mode based on the information of the parent ion, and then entered into the database as a basis for judging whether there are toxic substances.
[0036] b. Establishment of a simulated sample database containing toxic substances: Take a sample and add one of the common toxic substance standards with a final concentration of 0.3 μg / mL to it to obtain a target sample solution; use a syringe to draw 300 μL of the target sample solution and place it on a grooved sample sheet; use a methanol-acetonitrile mixture to flush the target solution into the capillary inlet, and then perform spray ionization and detection by an ion detector;
[0037] Finally, a spectral library of toxic substances was established, including the characteristic ion peak information of the parent ion and daughter ion corresponding to each toxic substance;
[0038] Analysis of the sample to be tested: 3000 μL of target sample solution is drawn using a syringe and placed on a grooved sample plate. The target solution is then flushed with a methanol-acetonitrile mixture to allow it to enter the capillary inlet, followed by spray ionization and detection. The sample is compared with the sample database containing toxic substances from step 1) to check for the presence of a parent ion peak corresponding to a toxic substance recorded in the toxic substance characteristic spectrum library. If no corresponding parent ion peak is found for any toxic substance, it indicates that the blood sample to be tested does not contain a toxic substance recorded in the toxic substance characteristic spectrum library. The detection is then completed, and a message indicating that no toxic substance information was found in the database is provided.
[0039] The poisons mentioned are quizalofop-P-ethyl, carbofuran, dimethoate, amitraz, omethoate, risperidone, chlorpromazine, clozapine, clonazepam, diazepam, acetaminophen, difenidol, chlorpheniramine, methamphetamine, methadone, noscapine, ketamine, amphetamine, AM-2201, and JWH-018.
[0040] The sample sheet is aluminum foil.
[0041] The groove depth is 0.5µm, and the spacing between adjacent grooves is 0.5mm;
[0042] Biological samples are human urine.
[0043] The ion trap mass spectrometry is a continuous injection ion trap mass spectrometry system equipped with an electrospray ionization source.
Claims
1. A method for detecting acute poisoning drugs in biological samples, characterized in that: An ion detection instrument equipped with an electrospray ion source is used; the ion detection instrument is an ion trap mass spectrometer. 200-1000 μL (preferably 400-600 μL) of biological sample is dropped into the groove of the sample sheet. Then, a methanol-acetonitrile mixture with a volume ratio of 1:10 to 1:1 (preferably 1:9) is used as an elution solution to rinse the sample droplets. Finally, the rinsing solution is ionized by an electrospray ionization source and then enters the detection instrument for detection. The grooved sample sheet refers to a sheet-like sample sheet with one or more grooves on its surface; The sample is placed in the groove, and the eluent is flushed by flowing through the sample along the length of the groove at a flow rate of 1 nL / min to 1 ml / min (preferably 10 uL / min) and a volume of 0.5 uL to 1 ml (preferably 5 uL). The results of the above tests were compared with the toxic substance characteristic spectrum library in the testing instrument to determine the type of poison in the sample.
2. The method according to claim 1, characterized in that: The specific analysis steps are as follows: 1) Establishment of a database of characteristic spectra of common toxic substances: a. The establishment of the toxic substance standard database employs an ion detection instrument equipped with an electrospray ionization source. Common toxic substance standards are diluted into three or more standard solutions of 0.1-1 μg / mL using a methanol-acetonitrile mixture. These standard solutions are then subjected to spray ionization within the electrospray ionization source. Ion detectors are then used to detect the toxic substances in both precursor and daughter ion modes. The spectra and peak positions of the precursor and daughter ions for different toxic substances are recorded and entered into the database as a basis for determining the presence of toxic substances. If an ion trap mass spectrometer is used as a detector, fragment ion information can be obtained by adjusting the working mode of the ion trap mass analyzer to the collision dissociation mode based on the information of the parent ion, and then entered into the database as a basis for judging whether there are toxic substances. b. Establishment of a simulated sample database containing toxic substances: Take a sample and add one of the common toxic substance standards with a final concentration of 0.1-1 μg / mL to it to obtain a target sample solution; use a syringe to draw 200-1000 μL of the target sample solution and place it on a grooved sample plate; use a methanol-acetonitrile mixture to flush the target solution into the capillary inlet, and then perform spray ionization and detection by an ion detector; Finally, a spectral library of toxic substances was established, including the characteristic ion peak information of the parent ion and daughter ion corresponding to each toxic substance; 2) Sample Analysis: Using a syringe, 200-1000 μL of target sample solution is drawn and placed on a grooved sample plate. The target solution is then flushed with a methanol-acetonitrile mixture to allow it to enter the capillary inlet, followed by spray ionization and detection. The sample is compared with the sample database containing toxic substances from step 1) to check for a corresponding parent ion peak for any toxic substance recorded in the toxic substance characteristic spectrum library. If no corresponding parent ion peak is found, it indicates that the blood sample does not contain any toxic substance recorded in the toxic substance characteristic spectrum library, and a message indicating no toxic substance information was found in the database is given. If a peak corresponds to a parent ion peak in the database, that parent ion is screened and subjected to collision dissociation mode detection to obtain its daughter ion information. The daughter ions are then compared: If the daughter ion matches the daughter ion of the corresponding parent ion in the database, toxic substance information is given; if no daughter ion corresponding to the parent ion in the database is detected, the detection is complete, and a message indicating no toxic substance information was found in the database is given.
3. The method according to claim 1 or 2, characterized in that: The poisons include one or more of the following four categories: pesticides, sedatives and hypnotics, therapeutic drugs, and narcotics. Specifically, it includes one or more of the following: quizalofop-P-ethyl, carbofuran, dimethoate, amitraz, omethoate, risperidone, chlorpromazine, clozapine, clonazepam, diazepam, acetaminophen, difenidol, chlorpheniramine, methamphetamine, methadone, noscapine, ketamine, amphetamine, AM-2201, and JWH-018.
4. The method according to claim 1 or 2, characterized in that: The sample sheet is one or more of aluminum foil, copper foil, and gold foil.
5. The method according to claim 1 or 2, characterized in that: The depth of the groove is 0.1µm to 5µm (preferably 0.5µm), and the spacing between adjacent grooves is 1µm to 1mm (preferably 0.5mm).
6. The method according to claim 1 or 2, characterized in that: Biological samples are one or more of the following: saliva, urine, or blood from a person or animal.
7. The method according to claim 1, characterized in that: Ion trap mass spectrometry is either discontinuous injection ion trap mass spectrometry or continuous injection ion trap mass spectrometry equipped with an electrospray ion source.