CN115389690A - Comprehensive identification method for benzotriazole ultraviolet absorber pollutants in environment - Google Patents
Comprehensive identification method for benzotriazole ultraviolet absorber pollutants in environment Download PDFInfo
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Abstract
A comprehensive identification method of benzotriazole ultraviolet absorber pollutants in the environment comprises the steps of obtaining a suspected target analysis database of benzotriazole ultraviolet absorber pollutants; performing liquid chromatography-mass spectrometry on an environmental sample to be detected to obtain data-dependent acquisition data and data-independent acquisition data; performing matching analysis on the data-dependent collected data based on the information of the suspected targeted compound, and determining the structure of the compound matched with the suspected targeted compound; extracting characteristic fragment ions of benzotriazole ultraviolet absorber-like contaminants from the data-independent collected data to extract candidate compounds based on the characteristic fragment ions; and analyzing chromatographic information and mass spectrum information related to the candidate compound in the data-dependent collected data to determine the structure of the candidate compound. The method realizes comprehensive identification of benzotriazole ultraviolet absorber pollutants in the environment, can be applied to various complex environment media, and has a wide application prospect.
Description
Technical Field
The invention relates to the field of environmental analytical chemistry, in particular to a comprehensive identification method for benzotriazole ultraviolet absorbent pollutants in the environment.
Background
Benzotriazole ultraviolet absorbers (BZT-UVs) are artificially synthesized organic compounds, and can absorb full spectrum ultraviolet rays in natural light, so that the Benzotriazole ultraviolet absorbers can be widely applied to the fields of plastics, coatings, textiles, paints, printing and dyeing, building materials, cosmetics and the like as chemical additives. During industrial production and daily use, BZT-UVs inevitably enter an environmental medium and are enriched in organisms. The environmental distribution and toxic effects of BZT-UVs have received extensive attention. The known BZT-UVs types in the current environment are extremely limited, and the comprehensive identification of the occurrence condition of the BZT-UVs in the environment medium is of great importance to the accurate evaluation of the health hazards and ecological risks of the BZT-UVs.
The analysis method is the key for comprehensively identifying the BZT-UVs. Traditionally, a targeted analysis method based on low-resolution mass spectrum is adopted in BZT-UVs environment monitoring, and depending on a real standard substance, screening and identification of unknown BZT-UVs are difficult to realize. The popularization and application of the high-resolution mass spectrum provide a technical means for comprehensively identifying BZT-UVs in a complex environment medium. A suspected targeted and non-targeted analysis method based on high-resolution mass spectrometry is one of effective strategies for guiding comprehensive identification of BZT-UVs homologues. However, how to screen and structurally identify the acquired massive high-resolution mass spectrum data based on a suspected targeted analysis method and a non-targeted analysis method is a technical problem which needs to be solved urgently.
Disclosure of Invention
In view of the above, it is a primary object of the present invention to provide a method for comprehensive identification of BZT-UVs-type contaminants in an environment, which is intended to at least partially solve at least one of the above mentioned technical problems.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a comprehensive identification method for BZT-UVs pollutants in an environment comprises the following steps: acquiring a suspected targeted analysis database of BZT-UVs pollutants, wherein the suspected targeted analysis database is constructed to store structural information and mass spectrum prediction information of a suspected targeted compound; performing liquid chromatography-mass spectrometry on an environmental sample to be detected, wherein fragment ions are respectively acquired in a data-dependent acquisition mode and a data-independent acquisition mode in the mass spectrometry so as to obtain data-dependent acquisition data and data-independent acquisition data of the environmental sample to be detected; performing matching analysis on the data-dependent acquired data based on the structural information and mass spectrum prediction information of the suspected targeted compound, and determining the determined or possible structure of the compound matched with the suspected targeted compound in the environmental sample to be tested; extracting characteristic fragment ions of a BZT-UVs class contaminant from the data-independent acquisition data to extract a candidate compound based on the characteristic fragment ions, wherein the candidate compound is distinguished from a compound matching the suspected targeting compound; analyzing chromatographic information and mass spectral information associated with the candidate compound in the data-dependent acquisition data to determine a likely structure of the candidate compound.
Based on the technical scheme, the comprehensive identification method of BZT-UVs pollutants in the environment has at least one or part of the following beneficial effects:
the method is based on the constructed suspected targeted analysis database of BZT-UVs pollutants, performs suspected targeted analysis on data-dependent acquired data in the liquid chromatography-mass spectrometry data of the environmental sample to be detected to obtain the determination or possible structure of a compound matched with the suspected targeted compound, performs non-targeted analysis on data-independent acquired data to obtain the possible structure of a candidate compound different from the suspected targeted compound, and realizes comprehensive identification and structure identification of BZT-UVs possibly existing in the environmental sample to be detected through complementation of two identification methods of the suspected targeted analysis and the non-targeted analysis, does not depend on a real standard product, can be implemented under the condition of lacking any reference compound information, can be applied to a complex environmental medium, and provides technical support for environmental monitoring.
Drawings
FIG. 1 is a flow chart of a method for comprehensively identifying BZT-UVs pollutants in the environment according to the invention.
FIG. 2 is a detailed flowchart of a method for comprehensively identifying BZT-UVs pollutants in the environment in embodiment 1 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the accompanying drawings in combination with the embodiments.
In the process of realizing the method, the technical difficulty that how to pertinently establish the suspected targeted analysis database in the suspected targeted analysis process is to adopt a suspected targeted analysis strategy to identify BZT-UVs is discovered; and non-targeted analysis does not have any reference compound information, and the application difficulty is to screen and structurally identify the acquired mass high-resolution mass spectrum data. The invention discloses a suspected targeted analysis database constructed based on the structural characteristics of BZT-UVs, which is combined with a recognition method of complementation of suspected targeted analysis and non-targeted analysis, and realizes comprehensive recognition of BZT-UVs homologues in a complex environment medium. It should be noted in advance that the database of suspected targets means that the database contains known BZT-UVs compounds that may exist in the environment, and the known BZT-UVs compounds that may exist in the environment are the suspected target compounds.
Specifically, according to some embodiments of the present invention, a method for comprehensive identification of BZT-UVs-like contaminants in an environment is provided, comprising the following steps A-E (FIG. 1).
Step A: and acquiring a suspected targeted analysis database of BZT-UVs pollutants, wherein the suspected targeted analysis database is constructed to store structural information and mass spectrum prediction information of a suspected targeted compound.
According to the embodiment of the invention, the suspected target analysis database in the step is constructed through the following steps A1-A3.
In step A1, all compounds having a 2-hydroxybenzotriazole structural fragment are screened from the public chemical database.
According to embodiments of the present invention, public chemical data includes, but is not limited to, china existing chemical lists (IECSCs), U.S. toxic quality control method lists (TSCAs), european union chemical registration, assessment, authorization and restriction lists (REACH), and canadian national materials lists (DSLs), among others.
According to an embodiment of the present invention, in order to facilitate accurate screening of compounds, step A1 specifically comprises: calculating the matching degree, namely the similarity, of the SMILES formula of the compound to be screened and the 2-hydroxybenzotriazole in the public chemical database based on a millet coefficient algorithm; and determining the compound to be screened as the compound with the 2-hydroxybenzotriazole structural fragment under the condition that the matching degree meets the preset condition.
Further optionally, the degree of matching is calculated by using a Python platform and a valley coefficient algorithm, and the molecular structural formula C of the 2-hydroxybenzotriazole 12 H 9 N 3 O, whose corresponding SMILES formula is C1= CC = C (= C1) N2N = C3C = CC3= N2) O. In the art, the SMILES formula is a form of describing a chemical structure with a character string, thereby converting a complex chemical structural formula into a character string form recognizable to a computer.
As a preferable example, in the case where the degree of matching is not less than 0.7, the compound to be screened corresponding to the degree of matching is determined as a compound having a 2-hydroxybenzotriazole structural fragment. Further preferably, after the computer screening based on the matching degree calculation, a manual check is further performed to improve the accuracy.
In step A2, the in vivo biotransformation products of the screening compound are predicted based on the in vivo phase I, phase II and phase III metabolic transformation pathways.
According to embodiments of the present invention, metabolic transformation pathways are determined based on metabolic transformation pathways of reported pollutants in vivo and BZT-UVs molecular structural features, such as those related to oxidation, hydrolysis, reduction, methylation, sulfation, acetylation, various binding reactions, and the like.
For example, the Compound discover (version 3.3) software from Thermo Fisher corporation can be used to predict the metabolic transformation products of BZT-UVs in phase I and phase II in organisms.
In step A3, a database of suspected targeted assays is constructed based on the screened compounds and the corresponding bioconversion products.
It is understood that the screening compound and the corresponding bioconversion product are regarded as suspected targeted compounds, and the suspected targeted analysis database contains structural information and mass spectrum prediction information of the screening compound and the corresponding bioconversion product.
According to the embodiment of the invention, the structural information comprises compound names and chemical formulas, and the mass spectrum prediction information comprises accurate mass numbers of the adducted ions and information of predicted secondary fragment ions. Alternatively, the compound name may be, for example, a full chemical name, an abbreviation chemical name, or the like; the formula may be, for example, of the formula, for example C 13 H 11 N 3 O and the like; the adduct ion accurate mass number may be, for example, a sodium added accurate mass number, a proton added accurate mass number, etc., with a proton added accurate mass number being more commonly used; predicting secondary fragment ion information may be predicted, for example, using MetFrag software.
And B: and performing liquid chromatography-mass spectrometry on the environmental sample to be detected, wherein the fragment ions are respectively acquired in a data-dependent acquisition mode and a data-independent acquisition mode in the mass spectrometry so as to obtain data-dependent acquisition data and data-independent acquisition data of the environmental sample to be detected.
According to the embodiment of the invention, in the step B, the ultra-high liquid chromatograph and the high resolution mass spectrometry system can be used for performing liquid chromatography-mass spectrometry on the environmental sample to be detected, so as to facilitate the analysis and detection of the complex environmental sample.
According to an embodiment of the invention, the liquid chromatography conditions are configured as a gradient elution procedure to better separate the environmental sample to be tested.
According to the embodiment of the invention, the data-dependent acquisition mode of mass spectrometry is a primary ion scanning mode and a secondary ion scanning mode under the condition of adopting an atmospheric pressure chemical ionization source, an electrospray ionization source or an atmospheric pressure chemical ionization source to generate positive ions; the data independent acquisition mode IS a primary ion scanning mode under the condition that an atmospheric pressure chemical ionization source, an electrospray ionization source or an atmospheric pressure chemical ionization source positive ion IS adopted under an in-source collision induced dissociation (IS-CID) mass spectrum mode. The ionization sources used in the data independent acquisition mode and the data dependent acquisition mode can be the same or different, and compared with the data dependent acquisition mode, the data independent acquisition mode adopts a full spectrum fragmentation mode in the source, so that all compound fragment information can be acquired, and the comprehensive identification of BZT-UVs pollutants is facilitated.
Further, the data-dependent acquisition data comprises a primary Mass Spectrum (MS) 1 ) And second order Mass Spectrum (MS) 2 )，MS 1 Obtained by electrostatic field orbital trap, the scanning range is mass-to-charge ratio m/z =100-1000 1 For obtaining information such as the mass number of the added ions, MS 2 Scanning range is MS dependent by high energy collisional dissociation (HCD) mode acquisition 1 The parent ion mass-to-charge ratio m/z of (2) is mainly used for acquiring secondary fragment ion information.
According to the embodiment of the invention, the data-dependent acquisition data needs to be subjected to high-resolution mass spectrum data deconvolution analysis, and the signal intensity threshold can be 10e4 or the like, for example.
Further, the data-independent acquisition data includes MS including information of all fragment ions 1 ，MS 1 Obtained by electrostatic field orbitrap, the scanning range is mass-to-charge ratio m/z =100-1000, and is mainly used for obtaining the signal intensity of characteristic fragment ions and the molecular formulas of all candidate compounds.
And C: and performing matching analysis on the data-dependent acquired data based on the structural information and mass spectrum prediction information of the suspected targeted compound, and determining the determined or possible structure of the compound matched with the suspected targeted compound in the environmental sample to be detected.
According to an embodiment of the present invention, step C is a suspected target analysis, and whether a precursor compound matching the suspected target compound exists in the environmental sample to be tested is analyzed to obtain a determined or possible structure of the precursor compound, which specifically includes substeps C1 to C3.
In the substep C1, matching the mass number of the adduct ions in the data-dependent acquired data with the accurate mass number of the adduct ions of the suspected targeted compound, and screening the suspected targeted compound to obtain a matched suspected precursor compound;
in substep C2, the secondary fragment ions in the data-dependent acquisition data are matched with the predicted secondary fragment ion information of the suspected precursor compound, and the matched precursor compound is obtained by screening from the suspected precursor compound;
in sub-step C3, the determined or likely structure of the precursor compound is determined based on the chromatographic retention behavior and the secondary fragment ions of the precursor compound, the precursor compound being a compound that matches the suspected targeting compound.
Through the substeps C1 to C3, the combined ion mass number and the secondary fragment ions are sequentially matched, so that whether a sample to be detected contains a suspected target compound in a suspected target database or not can be accurately determined, and the determination or possible structure of the precursor compound is provided by analyzing the chromatographic retention behavior (such as chromatographic retention time and the like) of a compound with successful characteristic matching and the mass spectrum fragmentation rule determined by the secondary fragment ions.
Step D: extracting characteristic fragment ions of the BZT-UVs class contaminant from the data-independent acquisition data to extract a candidate compound based on the characteristic fragment ions, wherein the candidate compound is distinguished from a compound matching the suspected targeting compound.
According to the embodiment of the invention, the step D is non-targeted analysis, the characteristic fragment ions of the BZT-UVs pollutants are generally secondary fragment ions shared by the BZT-UVs pollutants, so that theoretically, all BZT-UVs pollutants in the environmental sample to be detected can be determined by reverse inference based on the characteristic fragment ions, meanwhile, only the candidate compound different from the precursor compound in the step C is extracted in the step, the subsequent step analysis is carried out, and the suspected targeted analysis and the non-targeted analysis are coupled, so that the comprehensive identification of the BZT-UVs pollutants is realized while the analysis process is simplified.
According to an embodiment of the invention, this step D comprises in particular sub-steps D1 to D3.
In sub-step D1, an ion flow map (EIC) of characteristic fragment ions containing a BZT-UVs class of contaminants is extracted from the data-independent acquisition data.
In sub-step D2, the data is transferred from the MS corresponding to the EIC retention time 1 Extracting candidate molecular formulas corresponding to the characteristic fragment ions.
In sub-step D3, the candidate molecular formula that is distinguished from the precursor compound is determined as the molecular formula of the candidate compound.
Through the above substeps D1 to D3, the molecular formula of the candidate compound, such as C, which may correspond to the feature fragment ions is found based on the extracted feature fragment ions 20 H 25 N 3 O 2 And then step E is entered to infer possible structures based on the mass spectral data.
According to an embodiment of the invention, the characteristic fragment ions of a BZT-UVs-type contaminant may comprise known widely detected BZT-UVs secondary fragment ions, e.g. [ C ] 6 H 6 N 3 ] + (m/z = 120.0562) and [ C 12 H 10 N 3 O] + (m/z = 212.0824), and may also include characteristic fragment ions obtained by experimental analysis in the present invention, e.g. [ C ] 13 H 10 N 3 O] + (m/z = 224.0824) and [ C 15 H 14 N 3 O] + (m/z＝252.1137)。
Step E: and analyzing chromatographic information and mass spectrum information related to the candidate compound in the data-dependent acquired data to determine the possible structure of the candidate compound.
According to an embodiment of the present invention, this step E specifically comprises obtaining the chromatographic retention behavior and secondary fragment ions of the candidate compound from the data-dependent acquisition data to determine the likely structure of the candidate compound.
More specifically, the chromatographic retention behavior and the secondary fragment ions of the candidate compound are obtained from the data-dependent collected data based on the molecular formula of the candidate compound in the step D, and reasonable candidate compounds are identified by analyzing the chromatographic retention behavior and the mass spectrum fragmentation rule based on the secondary fragment ions, so that the possible structures of the candidate compounds are presumed.
According to an embodiment of the present invention, the method of the present invention further comprises determining characteristic fragment ions of a BZT-UVs class contaminant, specifically comprising the following steps F and G.
Step F: and performing liquid chromatography-mass spectrometry on a plurality of real standard substances existing in the environment in the suspected target analysis database in an IS-CID mass spectrometry mode. As a preferred embodiment, the real standard can select BZT-UVs which are found in the environment by means of text mining, and the text mining means includes but is not limited to key word extraction and the like.
G: and adjusting different taper hole voltages to analyze the characteristic fragment ion signals of the real standard substances so as to determine the optimal taper hole voltage and the corresponding characteristic fragment ions of BZT-UVs pollutants. It is understood that the characteristic fragment ions of the BZT-UVs class of contaminants are secondary fragment ions common to multiple authentic standards.
According to an embodiment of the present invention, the data-independent acquisition mode in step B is preferably performed under the optimal taper hole voltage condition.
According to the embodiment of the invention, the acquired real standard substance can be used for accurately determining the characteristic fragment ions of the BZT-UVs pollutants so as to accurately perform non-targeted analysis on the one hand, and can be used for performing targeted analysis on the compound matched with the real standard substance in the environment to be detected on the other hand.
Further, the method of the present invention further comprises performing targeted analysis on the environmental sample to be tested, specifically comprising step H: and performing matching analysis on the data-dependent acquired data based on the chromatographic retention behaviors and mass spectrum information of the plurality of real standards, and determining the targeted compounds and the content matched with the plurality of real standards. The quantitative analysis of the target compound in the environmental sample to be detected is realized through the target analysis.
According to the embodiment of the invention, the method can be applied to various complex environment media, and the environment sample to be detected comprises a water sample containing BZT-UVs, a solid sample and a biological sample; wherein, the water sample can be industrial sewage, inlet and outlet water of a sewage treatment plant, river water, surface water, seawater, drinking water, underground water and the like, the solid sample can be bottom mud, water sludge, sediment, soil, indoor dust, atmospheric particulates and the like of the sewage treatment plant, and the biological sample can be human breast milk, urine, serum, animal organs, fish, birds, sharks, mollusks, plants and the like.
The technical solution of the present invention will be described in detail below by referring to a plurality of specific examples. It should be noted that the following specific examples are only for illustration and are not intended to limit the invention.
In the following examples, some reagents and detection instruments are described below:
reagent: the 12 BZT-UVs authentic standards are listed in Table 1 below.
Liquid chromatography-mass spectrometer: ultra-high performance liquid chromatography and high resolution mass spectrometry (Ultimate-3000 liquid chromatography-orbital high resolution mass spectrometry, thermo Fisher, usa); and (3) chromatographic column: waters ACQUITY C18 (1.7 μm, 2.1X 100 mm).
Example 1
This example is an example of laboratory testing, focusing on the realization of identifying BZT-UVs present in a spiked sample using an established methodology, and includes the specific steps (fig. 2):
the method comprises the following steps: 16 BZT-UVs which are found in the environment and are shown in the table 1 are summarized in a text mining mode, and 12 BZT-UVs with higher detection rate and higher detection concentration are selected to purchase real standard products.
TABLE 1
Step two: based on an open chemical database containing IECSC, TSCA, REACH and DSL, the similarity between the compound to be screened and 2-hydroxybenzotriazole in the open chemical database is calculated by utilizing a Python platform and a cost coefficient algorithm, and a score threshold value is set to be 0.7, namely, the compound to be screened with the similarity more than or equal to 0.7 is determined to be the compound with the 2-hydroxybenzotriazole structural fragment. Since the true standard falls within the range of compounds screened in this step, the additional screening in this step resulted in 21 BZT-UVs homologs that were not purchased for the true standard.
Step three: based on 30 known pathways for metabolic conversion of contaminants in organisms as shown in Table 2, the I-phase and II-phase metabolic conversion products of BZT-UVs in organisms were predicted by using Compound resolver (version 3.3) software of Thermo Fisher corporation. The established pathway covers all 20 BZT-UVs bioconversion products currently known.
TABLE 2
Step four: and (3) constructing the BZT-UVs homologues obtained in the first step to the third step into a suspected target analysis database (namely an in-house database), wherein the suspected target analysis database comprises compound names, chemical formulas, accurate mass numbers of adducted ions and secondary fragment ion information predicted by MetFrag software.
Step five: and (3) continuously injecting 12 BZT-UVs with real standard substances selected in the step one in an IS-CID mass spectrum mode by a needle pump, adjusting the voltage of a taper hole to be 10, 20, 30, 40, 50, 60 and 70eV respectively, and comparing the signal intensity of the characteristic fragment ions of different BZT-UVs under series of taper hole voltages.
Preferably, 30eV IS the optimal cone-hole voltage for scanning characteristic fragment ions in the IS-CID mass spectrometry mode, so as to satisfy that all characteristic fragment ions have appropriate signal intensity.
Step six: 1mL of methanol solution containing 12 real standards with the concentration of 100 mu g/L is prepared, and then the sample injection analysis is carried out by a liquid chromatogram-mass spectrometer.
Liquid chromatography conditions: the temperature of the chromatographic column is 35 ℃; the mobile phase consisted of a methanol solution (A) containing 0.5mM ammonium acetate and an aqueous solution (B) containing 0.5mM ammonium acetate. The mobile phase gradient elution procedure was: first 70% A for 1min; increase a to 100% within 14 min; then 100% A was maintained for 5min; then reducing A to 70% in 0.1 min; finally 70% A for 4.9min; the flow rate of the mobile phase is 0.3mL/min; the injection volume was 5. Mu.L.
Mass spectrometryConditions are as follows: the ion source adopts an atmospheric pressure chemical ionization source positive ion mode; the spraying voltage is 3500V; the ion source temperature is 200 ℃, the ion transmission tube temperature is 350 ℃, and the atomization temperature is 400 ℃; the sheath gas, purge gas and auxiliary gas pressures were 35, 1 and 10Arb, respectively. Data dependent acquisition mode parameters: MS (Mass Spectrometry) 1 Obtained by electrostatic field orbitrap with a resolution of 120000 (m/z = 200), a scanning range of m/z =100-1000, a maximum injection time of 100ms, and an automatic gain control target of 3e6,s-lens RF of 60%. MS (Mass Spectrometry) 2 Acquisition in HCD mode, resolution 60000 (m/z = 200), collision energy setting at 10, 30, 50%, MS 1 Parent ions are isolated by a quadrupole, the width of an isolation window m/z =1, fragment ions are detected by an electrostatic field orbitrap, and the scanning range depends on the parent ions m/z. The data independent acquisition mode adopts an IS-CID mass spectrum mode, and the parameters are as follows: MS (Mass Spectrometry) 1 Obtained by electrostatic field orbitrap with a resolution of 120000 (m/z = 200), a scanning range of m/z =100-1000, a maximum injection time of 100ms, an automatic gain control target of 3e6, s-lens RF of 60%, a mass range of normal, a cone voltage of 30eV.
Step seven: the six step data dependent acquisition mode data were analyzed for suspected targets in Compound discover (version 3.3) software to identify BZT-UVs and predicted bioconversion products recorded in the list. The process is as follows: 1, deconvoluting high-resolution mass spectrum data, and setting a signal intensity threshold value to be 10e4; matching a suspected target database, wherein the matching comprises two mass spectrum data matching, namely adding ion mass number matching and secondary fragment ion matching of a primary mass spectrum, and the mass spectrum deviation is 5ppm, the isotope threshold is 75%, and the signal-to-noise ratio is 5 in the primary mass spectrum matching; and 3, analyzing the chromatographic retention behavior and mass spectrum fragmentation rule of the precursor compound with the successfully matched characteristics, and proposing a determined or possible structure.
Step eight: and (4) carrying out non-targeted analysis on the six-step data independent acquisition mode data in Xcalibur Qual Browser (version 4.0) software, and identifying BZT-UVs which are not recorded in a chemical industry list and do not belong to predicted biotransformation products, industrial intermediates or impurity classes. The process is as follows: 1, extracting 4 characteristic fragment ions, i.e. [ C ] 6 H 6 N 3 ] + (m/z＝120.0562)、[C 12 H 10 N 3 O] + (m/z＝212.0824)、[C 13 H 10 N 3 O] + (m/2 = 224.0824) and [ C 15 H 14 N 3 O] + An EIC of (m/z = 252.1137); 2, according to EIC chart retention time, from corresponding MS 1 Extracting characteristic fragment ions corresponding to the molecular formula of the candidate compound, wherein the molecular formula is different from the precursor compound which is successfully matched in the step seven; and 3, analyzing the chromatographic retention behavior and mass spectrum fragmentation rule of the candidate compound in the data-dependent acquisition mode based on the molecular formula, matching reasonable candidates based on mass spectrum data, and providing a determined or possible structure.
It will be appreciated that, furthermore, the BZT-UVs class contaminants identified in steps seven and eight may also be subjected to semi-quantitative analysis by means of structurally similar standards; the target analysis can also be performed on the environmental sample to be detected based on a plurality of real standard samples, and specifically, in this embodiment, the target analysis process is not repeated since the sample to be detected is the real standard sample.
Through the specific steps, all identification of 12 BZT-UVs in the standard solution is realized, the accuracy rate is 100%, the reliability of the identification method is shown, and the BZT-UVs existing in the sample can be accurately identified. The accurate identification of BZT-UVs in the sample is realized by the suspected targeted analysis in the step seven and the non-targeted analysis in the step eight, and the two identification methods are complementary, so that the comprehensive identification of BZT-UVs in the environment can be ensured.
Example 2
Similar to the procedure of example 1, the spiked methanol solution was replaced with spiked real environmental sediment samples and this example was designed to test the ability of the method to identify BZT-UVs in environmental samples in the presence of matrix interference.
The operation steps are basically the same as the example 1, and different from the example 1, in the sixth step, the pretreatment is carried out on the actual environment sample, and the pretreatment comprises the procedures of accelerated solvent extraction, gel permeation chromatography purification, silica gel column purification and rotary evaporation nitrogen blowing redissolution. The result shows that 12 marked BZT-UVs are completely identified, and the method is suitable for complex environment media and is slightly interfered by a matrix.
Example 3
This example is intended to test the overall ability of the method to identify BZT-UVs in environmental samples. The collected environmental sample is a chlamys farreri biological sample.
The operation steps are basically the same as the example 1, and different from the example 1, in the sixth step, the chlamys farreri sample is pretreated, and the pretreatment comprises the procedures of accelerated solvent extraction, gel permeation chromatography purification, silica gel column purification and rotary evaporation nitrogen blowing redissolution. The results show that 21 BZT-UVs are identified in total, and the 21 BZT-UVs comprise 10 targeted BZT-UVs (UV-P, UV-PS, UV-234, UV-320, UV-326, UV-327, UV-328, UV-329, UV-350 and UV-360), 5 biotransformation products (with structures shown in formulas I to V) and 4 impurity BZT-UVs (with structures shown in formulas 1 to 7). Wherein the BZT-UVs bioconversion products (UV-326-H and UV-327-CH) are dechlorinated and methylated 3 ) And the impurity BZT-UVs are found in an environmental medium for the first time by the identification method, which shows that the method can comprehensively identify the BZT-UVs in the environment and fills the technical blank in the field of the existing BZT-UVs environmental analysis chemistry.
Formula I to formula V: determination or probable Structure of identified BZT-UVs bioconversion products
BZT@m/z＝322：
BZT@m/z＝330：
BZT@m/z＝340：
BZT@m/z＝410：
Formula 1 to formula 7: determination or possible structure of identified impurity class BZT-UVs
Example 4
The purpose of this example was to test the high throughput identification capability of this method for BZT-UVs in large-scale environmental samples. The collected environment samples are 129 mollusk samples covering 9 cities (Dalian, yingkou, hulusi, north Daihe, tianjin, shou Guang, penglai, tobacco terrace and Weihai) in the Bohai area of China Ring.
The procedure is essentially the same as in example 1, except that in step six, pretreatment of the mollusk sample is carried out, including accelerated solvent extraction, purification by gel permeation chromatography, purification by silica gel column and rotary-steaming nitrogen-blowing redissolution procedures. The whole identification system completes analysis of all environmental samples within 24h, and the identified BZT-UVs with real standard substances are verified by the standard substances, so that the method can complete comprehensive identification of the BZT-UVs in the environment accurately, quickly and in high flux.
The results of the above embodiments comprehensively show that the comprehensive identification method of BZT-UVs pollutants in the environment has high accuracy, and the suspected targeted analysis method and the non-targeted analysis method are complementary; the method can be applied to various complex environment media, and is small in matrix interference; the identified compounds have comprehensive types, and known, unknown and transformed products can be identified efficiently; the method has the characteristics of rapidness and high flux, and can finish large-scale environmental sample identification in a short time. Therefore, the method has universal applicability and wide application prospect.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A comprehensive identification method for benzotriazole ultraviolet absorber pollutants in the environment comprises the following steps:
acquiring a suspected targeted analysis database of benzotriazole ultraviolet absorber pollutants, wherein the suspected targeted analysis database is constructed to store structural information and mass spectrum prediction information of suspected targeted compounds;
performing liquid chromatography-mass spectrometry on an environmental sample to be detected, wherein fragment ions are respectively acquired in a data-dependent acquisition mode and a data-independent acquisition mode in the mass spectrometry so as to obtain data-dependent acquisition data and data-independent acquisition data of the environmental sample to be detected;
performing matching analysis on the data-dependent acquired data based on the structural information and mass spectrum prediction information of the suspected targeted compound, and determining the determined or possible structure of the compound matched with the suspected targeted compound in the environmental sample to be tested;
extracting characteristic fragment ions of benzotriazole uv absorber-like contaminants from the data-independent acquisition data to extract a candidate compound based on the characteristic fragment ions, wherein the candidate compound is distinct from a compound that matches the suspected targeting compound;
analyzing chromatographic and mass spectral information associated with the candidate compound in the data-dependent acquisition to determine a likely structure of the candidate compound.
2. The comprehensive identification method according to claim 1, wherein said database of suspected targeted analyses is constructed by the steps of:
screening all compounds with 2-hydroxybenzotriazole structural fragments from public chemical databases;
predicting the biological transformation products of the screened compounds in the organisms on the basis of I-phase, II-phase and III-phase metabolic transformation pathways in the organisms;
constructing the suspected targeted analysis database based on the screening compound and the corresponding biotransformation product.
3. The comprehensive identification method of claim 2, wherein said screening all compounds having 2-hydroxybenzotriazole structural fragments from public chemical databases comprises:
calculating the matching degree of SMILES (Small ionic liquids) formulas of the compound to be screened and 2-hydroxybenzotriazole in the open chemical database based on a trough coefficient algorithm;
and under the condition that the matching degree meets a preset condition, determining the compound to be screened as the compound with the 2-hydroxybenzotriazole structural fragment.
4. The comprehensive identification method according to claim 1, wherein the data-dependent acquisition mode is a primary ion scanning mode and a secondary ion scanning mode under the condition of employing an atmospheric pressure chemical ionization source, an electrospray ionization source or an atmospheric pressure chemical ionization source with positive ions;
the data-independent acquisition mode is a primary ion scanning mode under the condition of positive ions in an in-source collision induced dissociation mass spectrum mode.
5. The comprehensive identification method according to claim 1 or 2, wherein the structural information includes a compound name and a chemical formula, and the mass spectrum prediction information includes an accurate mass number of an adduct ion and predicted secondary fragment ion information.
6. The comprehensive identification method according to claim 5, wherein said performing a match analysis on the data-dependent collected data based on the structural information and mass spectrum prediction information of the suspected targeting compound and determining the determined or probable structure of the compound in the environmental sample to be tested that matches the suspected targeting compound comprises:
matching the mass number of the adduct ions in the data-dependent collected data with the accurate mass number of the adduct ions of the suspected targeted compound, and screening the suspected targeted compound to obtain a matched suspected precursor compound;
matching the secondary fragment ions in the data-dependent acquisition data with the predicted secondary fragment ion information of the suspected precursor compound, and screening the suspected precursor compound to obtain a matched precursor compound;
determining the determined or likely structure of the precursor compound based on the chromatographic retention behavior and secondary fragment ions of the precursor compound, the precursor compound being a compound that matches the suspected targeting compound.
7. The comprehensive identification method according to claim 1, wherein said extracting characteristic fragment ions of benzotriazole uv absorber-based contaminants from the data-independent collection data so as to extract candidate compounds based on the characteristic fragment ions comprises:
extracting an ion flow graph containing characteristic fragment ions of benzotriazole ultraviolet absorber-like contaminants from the data-independent collected data;
extracting candidate molecular formulas corresponding to the characteristic fragment ions from a first-order mass spectrum corresponding to the retention time of the ion flow graph;
determining a candidate molecular formula that is distinct from the precursor compound as the molecular formula of the candidate compound.
8. The comprehensive identification method of claim 1, wherein said analyzing chromatographic and mass spectral information associated with the candidate compound in the data-dependent acquisition data to determine the likely structure of the candidate compound comprises:
obtaining chromatographic retention behavior and secondary fragment ions of the candidate compound from the data-dependent acquisition data to determine a likely structure of the candidate compound.
9. The comprehensive identification method according to claim 1, further comprising:
performing liquid chromatography-mass spectrometry on a plurality of real standards existing in the environment in the suspected target analysis database in an in-source collision induced dissociation mass spectrometry mode;
and adjusting different cone hole voltages to analyze the characteristic fragment ion signals of the real standards so as to determine the optimal cone hole voltage and the corresponding characteristic fragment ions of the benzotriazole ultraviolet absorbent pollutants.
10. The comprehensive identification method according to claim 9, further comprising:
and performing matching analysis on the data-dependent acquired data based on the chromatographic retention behaviors and the mass spectrum information of the real standards, and determining the target compounds and the content matched with the real standards.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116026960A (en) * | 2023-02-27 | 2023-04-28 | 北京师范大学 | Screening and identifying method and system for amino phenylsulfone pollutants in urban water body |
| CN116338069A (en) * | 2023-05-29 | 2023-06-27 | 国科大杭州高等研究院 | Organic phosphate high-throughput suspected target analysis method |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070093496A1 (en) * | 2003-10-20 | 2007-04-26 | Wai John S | Hydroxy pyridopyrrolopyrazine dione compounds useful as hiv integrase inhibitors |
| WO2015191999A1 (en) * | 2014-06-13 | 2015-12-17 | Waters Technologies Corporation | Analysis of complex biological matrices through targeting and advanced precursor and product ion alignment |
| CN106526016A (en) * | 2016-10-28 | 2017-03-22 | 陕西科技大学 | Ultra-high performance liquid chromatography-quadrupole electrostatic field orbital ion trap mass spectrometry screening method for plasticizer in milk and dairy products |
| US20170307589A1 (en) * | 2016-04-21 | 2017-10-26 | Nanjing University | Effect-directed identification of targeted and non-targeted androgen disruptors |
| CN109613253A (en) * | 2018-11-03 | 2019-04-12 | 杭州市农业科学研究院 | Utilize the method for the red cheek strawberry column cap differential protein of DDA-DIA interleaved acquisition quantitative screening |
| CN109870536A (en) * | 2017-12-05 | 2019-06-11 | 中国科学院大连化学物理研究所 | A kind of high covering iipidomics analysis method based on liquid chromatograph mass spectrography |
| CN110243985A (en) * | 2019-06-26 | 2019-09-17 | 南方科技大学 | A kind of Mass Spectrometry detection method of biomolecule |
| CN113552247A (en) * | 2021-06-18 | 2021-10-26 | 北京格竹生物科技有限公司 | Liquid chromatography-mass spectrometry non-target analysis method for unknown components of sample |
| WO2021217745A1 (en) * | 2020-04-26 | 2021-11-04 | 南京大学 | High-throughput screening method for non-target biomarker employing pollutant metabolic turbulence |
| CN113624869A (en) * | 2021-07-30 | 2021-11-09 | 上海市疾病预防控制中心 | Method for rapidly determining organic matter conversion substance in real sample with high flux and high accuracy |
| CN114577972A (en) * | 2020-11-30 | 2022-06-03 | 中国科学院大连化学物理研究所 | Protein marker screening method for body fluid identification |
| WO2022121055A1 (en) * | 2020-12-12 | 2022-06-16 | 广州禾信仪器股份有限公司 | Metabolomics-based method and apparatus for physiological prediction, computer device, and medium |
-
2022
- 2022-09-27 CN CN202211194533.9A patent/CN115389690B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070093496A1 (en) * | 2003-10-20 | 2007-04-26 | Wai John S | Hydroxy pyridopyrrolopyrazine dione compounds useful as hiv integrase inhibitors |
| WO2015191999A1 (en) * | 2014-06-13 | 2015-12-17 | Waters Technologies Corporation | Analysis of complex biological matrices through targeting and advanced precursor and product ion alignment |
| US20170307589A1 (en) * | 2016-04-21 | 2017-10-26 | Nanjing University | Effect-directed identification of targeted and non-targeted androgen disruptors |
| CN106526016A (en) * | 2016-10-28 | 2017-03-22 | 陕西科技大学 | Ultra-high performance liquid chromatography-quadrupole electrostatic field orbital ion trap mass spectrometry screening method for plasticizer in milk and dairy products |
| CN109870536A (en) * | 2017-12-05 | 2019-06-11 | 中国科学院大连化学物理研究所 | A kind of high covering iipidomics analysis method based on liquid chromatograph mass spectrography |
| CN109613253A (en) * | 2018-11-03 | 2019-04-12 | 杭州市农业科学研究院 | Utilize the method for the red cheek strawberry column cap differential protein of DDA-DIA interleaved acquisition quantitative screening |
| CN110243985A (en) * | 2019-06-26 | 2019-09-17 | 南方科技大学 | A kind of Mass Spectrometry detection method of biomolecule |
| WO2021217745A1 (en) * | 2020-04-26 | 2021-11-04 | 南京大学 | High-throughput screening method for non-target biomarker employing pollutant metabolic turbulence |
| CN114577972A (en) * | 2020-11-30 | 2022-06-03 | 中国科学院大连化学物理研究所 | Protein marker screening method for body fluid identification |
| WO2022121055A1 (en) * | 2020-12-12 | 2022-06-16 | 广州禾信仪器股份有限公司 | Metabolomics-based method and apparatus for physiological prediction, computer device, and medium |
| CN113552247A (en) * | 2021-06-18 | 2021-10-26 | 北京格竹生物科技有限公司 | Liquid chromatography-mass spectrometry non-target analysis method for unknown components of sample |
| CN113624869A (en) * | 2021-07-30 | 2021-11-09 | 上海市疾病预防控制中心 | Method for rapidly determining organic matter conversion substance in real sample with high flux and high accuracy |
Non-Patent Citations (5)
| Title |
|---|
| TING RUAN 等: "Analytical methodology for identification of novel per- and polyfluoroalkyl substances in the environment", TRENDS IN ANALYTICAL CHEMISTRY, vol. 95, pages 122 - 131 * |
| 勾新磊 等: "高效液相色谱-高分辨质谱法快速筛查食品塑料包装袋中的5种苯并三唑类紫外吸收剂", 分析仪器, no. 5, pages 31 - 36 * |
| 张美娟 等: "高分辨质谱-非信息依赖数据采集-后靶向筛查策略在不明原因食物中毒鉴定中的应用", 分析化学研究报告, vol. 46, no. 2, pages 157 - 164 * |
| 王成云 等: "纺织品中紫外线吸收剂的快速筛查和确证", 深圳大学学报(理工版), vol. 34, no. 3, pages 322 - 330 * |
| 雷汝清 等: "超高效液相色谱-四极杆飞行时间质谱法非靶向筛查西红柿中三唑类杀菌剂", 色谱, vol. 38, no. 7, pages 805 - 816 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116026960A (en) * | 2023-02-27 | 2023-04-28 | 北京师范大学 | Screening and identifying method and system for amino phenylsulfone pollutants in urban water body |
| CN116026960B (en) * | 2023-02-27 | 2023-06-16 | 北京师范大学 | Screening and identifying method and system for amino phenylsulfone pollutants in urban water body |
| CN116338069A (en) * | 2023-05-29 | 2023-06-27 | 国科大杭州高等研究院 | Organic phosphate high-throughput suspected target analysis method |
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