High resolution mass spectrometry method for simultaneous determination of multiple psychoactive substances and their metabolites
By combining UHPLC-Q-Orbitrap HRMS with a dedicated database, the problem of rapid and accurate screening of various psychoactive substances and their metabolites in existing technologies has been solved, achieving high-throughput, matrix-resistant, and highly sensitive analytical results.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- YUNNAN POLICE COLLEGE
- Filing Date
- 2026-03-16
- Publication Date
- 2026-05-29
AI Technical Summary
Existing detection technologies are insufficient for rapidly and accurately screening multiple traditional drugs and new psychoactive substances and their metabolites simultaneously. In particular, they suffer from problems such as identification lag, lack of standard substances, insufficient high-throughput screening capabilities, and strong interference from complex matrices.
Ultra-high performance liquid chromatography (UHPLC-Q-Orbitrap HRMS) was used for separation and data-dependent acquisition (DDA) to construct a dedicated database containing accurate precursor ion mass, chromatographic retention time, and multi-level characteristic fragment ion information. Through strict qualitative judgment criteria, a variety of psychoactive substances and their metabolites were identified.
It achieves high-throughput, rapid, and accurate screening of up to 75 psychoactive substances and their metabolites, and has good resistance to matrix interference and sensitivity, making it suitable for complex sample analysis such as drug enforcement and wastewater monitoring.
Smart Images
Figure CN122109376A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of analytical chemistry, specifically to a high-resolution mass spectrometry method for the simultaneous determination of multiple psychoactive substances and their metabolites. Background Technology
[0002] The abuse of psychoactive substances is a major global public safety issue, posing a persistent threat to public health and social stability. Currently, it faces three main challenges: First, the abuse of traditional drugs such as heroin, cocaine, and methamphetamine remains widespread; second, new psychoactive substances (NPS), such as synthetic cannabinoids, synthetic cathinones, and fentanyl analogues, are constantly emerging, with rapidly changing chemical structures and a wide variety, and national control measures often lag behind their emergence, resulting in "detection gaps" in law enforcement and monitoring; third, the need for preventative identification of potential psychoactive substances is increasingly prominent. New compounds with psychoactive potential may emerge in pharmaceutical research and academic studies, and may enter illicit channels before being controlled, thus necessitating the establishment of "pre-detection" capabilities to achieve early warning and risk assessment.
[0003] At the detection technology level, current methods primarily rely on chromatography-mass spectrometry (GC-MS). Gas chromatography-mass spectrometry (GC-MS) is effective for volatile substances, but requires complex derivatization steps for thermally unstable or highly polar compounds. Liquid chromatography-tandem triple quadrupole mass spectrometry (LC-QqQMS), while highly sensitive and specific, is a "targeted" analysis requiring pre-setting of monitoring ion pairs, making it difficult to handle high-throughput screening of unknown or novel structural variants. High-resolution mass spectrometry (HRMS), especially the electrostatic orbital trap technique, offers high quality, precision, and resolution, combined with data-dependent acquisition (DDA) mode, possessing the potential for both non-targeted discovery and targeted confirmation. However, there are still a series of technical bottlenecks in applying it to high-confidence screening of multiple categories of psychoactive substances in complex real samples such as drug control and environmental monitoring: (1) lack of efficient and universal pretreatment process to extract compounds with large differences in properties and purify the matrix at the same time; (2) chromatographic separation conditions are difficult to achieve effective separation of high-throughput compounds; (3) lack of systematic optimization of mass spectrometry parameter settings to ensure data quality and acquisition efficiency; (4) lack of reliable database containing multi-dimensional information such as retention time and multi-level fragment ions.
[0004] Therefore, existing detection systems generally suffer from common problems when dealing with traditional drugs, rapidly evolving NPS (Net Producer Status Indicators), and potential psychoactive substances not yet listed as controlled substances. These problems include identification lag, lack of standard materials, insufficient high-throughput screening capabilities, and strong interference from complex matrices. To cover the full spectrum of identification needs, from known to unknown, and from controlled to potential risk substances, there is an urgent need to develop a rapid, sensitive, broad-spectrum analytical method with high confidence in identification capabilities.
[0005] Therefore, developing a high-resolution mass spectrometry method that can overcome the above difficulties and enable rapid and accurate determination and identification of up to dozens of psychoactive substances and their metabolites, including traditional drugs and NPS, is of significant technical value and has an urgent application need. Summary of the Invention
[0006] The purpose of this application is to provide a high-resolution mass spectrometry method for the simultaneous determination of multiple psychoactive substances and their metabolites, in order to solve the problems in the prior art.
[0007] To achieve the above objectives, embodiments of this application provide a high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites, comprising:
[0008] S1. Sample pretreatment: Pretreatment of complex matrix samples to obtain the test solution;
[0009] S2. Data Acquisition: After the test liquid is separated by ultra-high performance liquid chromatography, Full MS / dd-MS² mass spectrometry data is acquired using a mass spectrometer in data-dependent acquisition mode.
[0010] S3. Database construction and comparison: Based on the standard information of various target psychoactive substances and their metabolites, a dedicated database containing the precise precursor ion mass, chromatographic retention time and multi-level characteristic fragment ion information of each target compound is constructed. The mass spectrometry data collected in step S2 is compared and analyzed with the information in the dedicated database to obtain the comparison results.
[0011] S4. Qualitative determination: Based on the comparison results in step S3, the presence or absence of the target compound in the complex matrix is determined and identified according to the preset qualitative determination criteria.
[0012] Optionally, the psychoactive substances and their metabolites include synthetic cannabinoids, imidazoles, phenethylamines, synthetic cathinones, fentanyls, and ketamines.
[0013] Optionally, the complex matrix sample may be a seized solid / powder suspected sample or an environmental sewage sample.
[0014] Optionally, the pretreatment includes steps of dissolving, extracting and diluting solid samples, and steps of filtering and solid-phase extraction of liquid samples.
[0015] Optionally, in step S2, the separation of the ultra-high performance liquid chromatography uses a C18 column, and the mobile phase is a 0.1% formic acid aqueous solution and an acetonitrile solution, with gradient elution.
[0016] Optionally, in step S2, the resolution of the Full MS scan is not less than 70,000 FWHM, and the resolution of the dd-MS² scan is not less than 17,500 FWHM.
[0017] Optionally, in step S3, the dedicated database also includes ionization mode information, precise mass number of characteristic fragment ions, and retention time of each target compound.
[0018] Optionally, in step S4, the qualitative determination criteria include: the deviation between the precise mass number of the first-order mass spectrometer of the compound to be tested and the mass number of the precursor ion in the database is less than 5 ppm, the deviation between the retention time and the reference value in the database is less than 2.5%, and at least one characteristic fragment ion matches the dedicated database.
[0019] Optionally, in step S2, the selection of the precursor ion in the data-dependent acquisition mode is based on a preset list containing the precise precursor ion mass numbers of various target compounds.
[0020] The embodiments of this application have the following advantages:
[0021] High throughput and broad spectrum: By optimizing chromatographic conditions and adopting the DDA acquisition mode of high-resolution mass spectrometry, the method enables the simultaneous determination of 75 psychoactive substances and their metabolites covering more than six major categories with a single injection (the biomarker cotinine was added because this method can be applied to wastewater detection), which greatly improves the analytical throughput and screening range.
[0022] High-confidence identification: Based on triple or multiple comparisons of precise mass number, retention time and multi-level fragment ion information, coupled with strict judgment criteria, it significantly improves the accuracy and reliability of qualitative identification and effectively reduces the risk of false positives caused by complex matrices.
[0023] Strong resistance to matrix interference: High-resolution and accurate mass measurement can effectively distinguish target compounds from matrix background interference ions; the universal and efficient sample pretreatment process further reduces matrix effects, making the method applicable to the direct analysis of various complex matrices such as seizures and sewage.
[0024] Good sensitivity and reproducibility: The method can reliably detect the target analyte at concentration levels as low as ng / mL or even sub-ng / mL, and the retention time and mass spectrometry response are reproducible, meeting the requirements for quantitative and semi-quantitative analysis.
[0025] Good scalability and foresight: The established database and analysis framework are easy to expand. When new substances appear, their standard information can be easily added to the database, enabling the method to quickly adapt to the rapid changes in NPS.
[0026] The method proposed in this application provides a powerful technical tool for drug enforcement agencies to conduct rapid physical evidence identification of seized drugs and to conduct precise monitoring and early warning of regional drug situations based on wastewater epidemiology. It can also be widely applied in related fields such as forensic toxicology, clinical toxicology, and doping detection. Attached Figure Description
[0027] To more clearly illustrate the embodiments of this application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.
[0028] Figure 1 A flowchart illustrating a high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites, provided for at least one embodiment of this application;
[0029] Figure 2 TIC chromatograms of 75 psychoactive substances and their metabolites provided for at least one embodiment of this application, which is a high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites.
[0030] Figure 3 Chromatograms, full-range mass spectrometry (FMS) and dd-MS² of a target compound (taking O6-monoacetylmorphine as an example) for a high-resolution mass spectrometry method for the simultaneous determination of multiple psychoactive substances and their metabolites provided in at least one embodiment of this application.
[0031] Figure 4 The isomers of isopyramidase and isopyramidase are shown in the following chromatograms (A), full MS (B), and dd-MS² (C) of isopyramidase, and full MS (D) and dd-MS² (E) of isopyramidase, which are provided for at least one embodiment of this application for a high-resolution mass spectrometry method for the simultaneous determination of multiple psychoactive substances and their metabolites. Detailed Implementation
[0032] The following specific embodiments illustrate the implementation of this application. Those skilled in the art can easily understand other advantages and effects of this application from the content disclosed in this specification. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.
[0033] It should be noted that the steps in the claims and description of this application may be performed substantially in parallel or in reverse order under appropriate circumstances, depending on the function involved.
[0034] Furthermore, the technical features involved in the different embodiments of this application described below can be combined with each other as long as they do not conflict with each other.
[0035] This application provides a high-resolution mass spectrometry method for simultaneously determining multiple (e.g., 75 existing) psychoactive substances and their metabolites, referencing... Figure 1 , Figure 1 The flowchart illustrates a high-resolution mass spectrometry method for the simultaneous determination of multiple psychoactive substances and their metabolites, provided in at least one embodiment of this application. It should be understood that the method may further include additional boxes not shown and / or the boxes shown may be omitted; the scope of this application is not limited in this respect. The method includes:
[0036] (1) Sample pretreatment: For different types of complex matrix samples (such as seized suspicious powders, tablets or environmental sewage), an efficient and universal pretreatment process is adopted. For solid samples, the process includes grinding, weighing and methanol dissolution; for liquid sewage samples, the process includes filtration, solid phase extraction (SPE) enrichment and purification, rotary evaporation and resolution, etc., to finally obtain a test solution suitable for instrument analysis.
[0037] (2) Chromatography-Mass Spectrometry (GC-MS): The analyte was injected into an ultra-high performance liquid chromatography (UHPLC) system for separation. Chromatographic separation used a reversed-phase C18 column, with a mobile phase of an aqueous solution containing 0.1% formic acid (phase A) and an acetonitrile solution (phase B), employing a gradient elution program. The separated components were then detected by a quadrupole / electrostatic field orbital trap high-resolution mass spectrometer (Q-Orbitrap HRMS). Mass spectrometry data acquisition employed a data-dependent acquisition (DDA) mode, specifically Full MS / dd-MS² mode: First, a full scan (Full MS) was performed to acquire the precise mass numbers of all ions at high resolution (e.g., ≥70,000 FWHM); then, based on a pre-defined trigger list containing the mass numbers of multiple target precursor ions, precursor ions with intensities exceeding a threshold were dynamically selected for fragmentation, followed by a second-stage high-resolution scan (dd-MS², resolution e.g., ≥17,500 FWHM) to obtain characteristic fragment ion information. Reference Figure 2 and Figure 3 , Figure 2 TIC plots for 75 psychoactive substances and their metabolites. Figure 3 Chromatograms, full MS, and dd-MS² of the target compound (using O6-monoacetylmorphine as an example).
[0038] (3) Database construction: Pure standard solutions of various target psychoactive substances and their metabolites (at least covering synthetic cannabinoids, imidyl esters, phenethylamines, synthetic cathinones, fentanyl derivatives, and ketamine) were injected under analytical conditions identical to those of the actual samples, and their precise precursor ion masses ([M+H)) were collected. + The system collects retention time (RT), as well as the precise mass number and relative abundance ratio of multi-level characteristic fragment ions, to construct a dedicated high-resolution mass spectrometry database (dedicated database) containing this multi-dimensional information.
[0039] (4) Data Processing and Qualitative Determination: The mass spectrometry data (including primary and secondary precise mass numbers and retention times) collected from the actual sample are compared with the dedicated database. Determination is made according to the established strict qualitative criteria: the criteria require that the mass deviation between the precise mass number of the precursor ion of the target peak and the database reference value be less than 5 ppm (parts per million); the deviation between its chromatographic retention time and the database reference retention time be within 2.5%; and at least one characteristic fragment ion must be matched in its secondary mass spectrum (preferably at least two characteristic fragment ions). Only components that meet all the above conditions can be confirmed as the corresponding target compound. If there are two or more chromatographic peaks with the same primary precise mass number but different retention times, it is generally considered that isomers exist. The specific substance is determined based on the characteristic fragment ions in the secondary mass spectrum, as follows: Figure 4 As shown, Figure 4 Chromatograms of isopramipexole and isopramipexole (A), precise first-order mass spectrum (Full MS) (B) and second-order mass spectrum (dd-MS²) (C) of isopramipexole, and precise first-order mass spectrum (Full MS) (D) and second-order mass spectrum (dd-MS²) (E) of isopramipexole.
[0040] The method described in this application exhibits high sensitivity for the detection of the vast majority of target compounds, with a limit of detection (LOD) of 0.05-10 ng / mL in complex matrices. This method is suitable for high-throughput screening and confirmation in the identification of seized drugs during drug enforcement, environmental wastewater toxicity monitoring, forensic identification, or clinical toxicology analysis.
[0041] In summary, this application discloses a high-resolution mass spectrometry (HMS) method for the simultaneous determination of 75 psychoactive substances and their metabolites in complex matrices using ultra-high performance liquid chromatography-quadrupole / electrostatic field orbital trap HRMS (UHPLC-Q-Orbitrap HRMS). Specifically, it relates to a rapid, highly sensitive, and highly selective UHPLC-orbitrap HRMS method capable of simultaneously determining and identifying up to 75 psychoactive substances and their metabolites across different categories in complex matrices. This method employs an efficient and universal sample pretreatment process, combined with a Full MS / dd-MS² data-dependent HMS acquisition mode, to construct a dedicated database containing accurate precursor ion mass, chromatographic retention time, and multi-level characteristic fragment ion information. By systematically optimizing chromatographic and mass spectrometric parameters and setting strict qualitative judgment criteria, this method achieves simultaneous and highly selective determination and identification of 75 psychoactive substances and their metabolites across multiple categories, including imidyl esters, synthetic cannabinoids, phenethylamines, synthetic cathinones, fentanyl derivatives, and ketamines, in a single injection. The method exhibits high sensitivity, reliably detecting the vast majority of target compounds within the concentration range of 0.05–10 ng / mL. Validated by actual seizures of suspicious samples and environmental wastewater samples, this method demonstrates simple pretreatment, rapid analysis speed, high specificity, and outstanding resistance to matrix interference. This method is suitable for the rapid identification of seized drugs in drug enforcement, drug monitoring in wastewater epidemiology, and can be extended to high-throughput analysis of multiple psychoactive substances in complex biological or environmental samples in fields such as forensic medicine and clinical toxicology.
[0042] The present application will be further described in detail below with reference to specific embodiments. The following embodiments are for illustrative purposes only and are not intended to limit the scope of the application.
[0043] Example 1: Instrumentation Methods and Database Construction
[0044] Instruments and Reagents: A Thermo Scientific ultra-high performance liquid chromatography system coupled with a quadrupole-electrostatic field orbital trap high-resolution mass spectrometer (Vanquish Flex UHPLC-Q Exactive Focus HRMS) was used. The column was a Proshell 120 EC-C18 (100 mm × 2.1 mm, 2.7 μm, Agilent). Mobile phase A was 0.1% formic acid aqueous solution, and mobile phase B was acetonitrile solution, with a flow rate of 0.4 mL / min. The injection volume was 1 μL. The gradient elution program of the mobile phase was as follows: the proportion of acetonitrile was maintained at 10% from 0 to 0.5 min, increased to 95% from 0.5 to 15.0 min, maintained at 95% from 15.0 to 15.5 min, decreased to 10% from 15.5 to 16.0 min, and maintained at 10% for 20 min to reequilibrate the column before the next injection.
[0045] Mass spectrometry parameters: Electrospray ionization (ESI) source, positive ion mode. Full MS scan: resolution 70,000 FWHM, scan range m / z 70-1000, maximum injection time 100 ms. dd-MS² scan: resolution 17,500 FWHM, isolation window 1.0 m / z, normalized collision energies (NCE) 20, 30, and 40 eV, maximum injection time 50 ms, automatic gain control (AGC) 1.0e3, dynamic exclusion 3.0 s.
[0046] Database Construction: Precisely prepare mixed standard solutions (1 μg / mL, methanol) for 75 target compounds (listed in Table 1). Inject each compound individually (1 μL injection volume), collecting retention times and first-order exact mass numbers ([M+H)). + The data includes the secondary fragment ion spectra. Using TraceFinder software, characteristic fragment ions for each compound were manually extracted and identified (usually 2-3 ions with high abundance and strong characteristics were selected). All information (compound name, molecular formula, theoretical mass, measured mass, retention time, and m / z of characteristic fragment ions) was compiled into a local database (Table 1).
[0047] Table 1: UPLC-Q-Orbitrap HRMS analysis parameters of 75 psychoactive substances and their metabolites
[0048]
[0049]
[0050]
[0051]
[0052]
[0053] Example 2: Analysis of seized suspicious powder samples
[0054] Sample pretreatment: Accurately weigh 10 mg of the seized suspected powder sample into a 15 mL centrifuge tube, add 10 mL of methanol, vortex to mix, and then sonicate for 30 min. Centrifuge at 12000 rpm for 10 min, collect the supernatant, filter through a 0.22 μm PVDF membrane, dilute 1000 times, and transfer to a vial for analysis by UHPLC-Q-Orbitrap HRMS.
[0055] Sample analysis and identification: The sample solution was analyzed according to the conditions of Example 1. The data processing software automatically retrieved and compared the acquired mass spectra with the dedicated database constructed in Example 1.
[0056] Results: The software report detected multiple chromatographic peaks. Taking the peak with a retention time of 2.45 min as an example, its first-order exact mass number was 328.15140 ([M+H]⁺), with a mass deviation of -0.5 ppm from that of O6-monoacetylmorphine (exact mass 328.15158) in the database. The retention time of O6-monoacetylmorphine in the database was 2.48 min, with a time deviation of -1.2%. Its second-order mass spectrum showed two major fragment ions, m / z 165.06882 and 211.07430, which matched the characteristic fragment ions of this compound in the database: 165.06816 (mass deviation 3.9 ppm) and 211.07387 (mass deviation 2.0 ppm). According to the judgment criteria (mass deviation < 5 ppm, RT deviation < ± 2.5%, and matching ≥ 2 fragment ions), this peak was confirmed as O6-monoacetylmorphine.
[0057] Using this method, compounds such as codeine, noscapine, and caffeine were simultaneously identified.
[0058] Example 3: Analysis of environmental wastewater samples
[0059] Sample pretreatment: 1800 mL of environmental wastewater sample was taken and filtered through a vacuum filter membrane. Enrichment and purification were performed using an HLB solid-phase extraction column (50 mg / 3 mL) and a high-throughput automated solid-phase extraction system (Fotector Plus, Ruike Group (Xiamen) Co., Ltd.): the column was activated sequentially with 5 mL of methanol and 5 mL of ultrapure water; the sample was loaded; and eluted with 4 mL of methanol. The eluent was rotary evaporated to near dryness in a vacuum centrifuge (Beijing Jiam Technology Co., Ltd., JM50) at 1600 r / min and 45℃, reconstituted with 1 mL of methanol, vortexed, and filtered through a 0.22 μm filter membrane for analysis.
[0060] Sample analysis and identification: The sample solution was analyzed under the conditions of Example 1 and compared with a dedicated database.
[0061] Results: Multiple psychoactive substances, including caffeine, methamphetamine, lidocaine, and cotinine, as well as the metabolic marker cotinine, were successfully identified in wastewater. Even at target concentrations in the ng / L range, reliable identification was achieved in complex wastewater matrix backgrounds through precise mass filtering and characteristic fragment ion matching by high-resolution mass spectrometry, demonstrating the method's excellent sensitivity and anti-interference capability.
[0062] Note that, unless otherwise explicitly stated, all features disclosed in this specification (including any appended claims, abstract, and drawings) may be replaced by alternative features for achieving the same, equivalent, or similar purpose. Therefore, unless explicitly stated otherwise, each disclosed feature is merely one example of a set of equivalent or similar features. Where used, "further," "preferably," "even further," and "more preferably" are simply starting points for describing another embodiment based on the foregoing embodiments, the combination of which with the foregoing embodiments constitutes the complete configuration of another embodiment. Any combination of several "further," "preferably," "even further," or "more preferably" settings following the same embodiment constitutes yet another embodiment.
[0063] In the implementation of functions and steps, the corresponding functions and steps in the various embodiments may occur in a different order than those shown. For example, two consecutive functions and steps may actually be executed or implemented substantially in parallel, and they may sometimes be executed or implemented in reverse order, depending on the functions involved.
[0064] Although this application has been described in detail above with general descriptions and specific embodiments, some modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of this application fall within the scope of protection claimed in this application.
Claims
1. A high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites, characterized in that, include: S1. Sample pretreatment: Pretreatment of complex matrix samples to obtain the test solution; S2. Data Acquisition: After the test liquid is separated by ultra-high performance liquid chromatography, Full MS / dd-MS² mass spectrometry data is acquired using a mass spectrometer in data-dependent acquisition mode. S3. Database construction and comparison: Based on the standard information of various target psychoactive substances and their metabolites, a dedicated database containing the precise precursor ion mass, chromatographic retention time and multi-level characteristic fragment ion information of each target compound is constructed. The mass spectrometry data collected in step S2 is compared and analyzed with the information in the dedicated database to obtain the comparison results. S4. Qualitative determination: Based on the comparison results in step S3, the presence or absence of the target compound in the complex matrix is determined and identified according to the preset qualitative determination criteria.
2. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, The psychoactive substances and their metabolites include synthetic cannabinoids, imidazoles, phenethylamines, synthetic cathinones, fentanyls, and ketamines.
3. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, The complex matrix sample is a seized solid / powder suspected sample or an environmental sewage sample.
4. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, The pretreatment includes steps of dissolving, extracting and diluting solid samples, and steps of filtering and solid-phase extraction of liquid samples.
5. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, In step S2, the separation by ultra-high performance liquid chromatography uses a C18 column, with a mobile phase of 0.1% formic acid aqueous solution and acetonitrile solution, and gradient elution is performed.
6. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, In step S2, the resolution of the Full MS scan is not less than 70,000 FWHM, and the resolution of the dd-MS² scan is not less than 17,500 FWHM.
7. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, In step S3, the dedicated database also includes ionization mode information, precise mass number of characteristic fragment ions, and retention time for each target compound.
8. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, In step S4, the qualitative determination criteria include: the deviation between the precise mass number of the first-order mass spectrometer of the compound to be tested and the mass number of the precursor ion in the database is less than 5 ppm, the deviation between the retention time and the reference value in the database is less than 2.5%, and at least one characteristic fragment ion matches the dedicated database.
9. The high-resolution mass spectrometry method for simultaneously determining multiple psychoactive substances and their metabolites according to claim 1, characterized in that, In step S2, the selection of the precursor ion in the data-dependent acquisition mode is based on a preset list containing the precise precursor ion mass numbers of various target compounds.