A stable isotope-labeled APMP derivative reagent and double library system-based aldehyde compound targeted-non-targeted synergistic analysis method

Abstract

This invention belongs to the field of environmental analytical chemistry and mass spectrometry, specifically relating to a targeted-non-targeted synergistic analysis method for aldehyde compounds based on stable isotope-labeled APMP derivatization reagents and a dual-library system. This method synthesizes d0 and d3 isotope-labeled APMP reagents to derivatize aldehyde compounds, and combines this with liquid chromatography-mass spectrometry (LC-MS) to achieve highly sensitive detection. Furthermore, a self-built dual-library system containing both derivatized and non-derivatized libraries is constructed, combining targeted quantification and non-targeted screening to achieve synergistic identification and quantitative analysis of known and unknown aldehyde compounds. Isotope internal standard correction and library matching verification improve the accuracy and reliability of the detection. This invention overcomes the limitations of traditional methods in terms of limited detection coverage and insufficient non-targeted quantitative analysis capabilities, making it suitable for high-throughput analysis of aldehyde compounds in complex environmental samples.

Description

Technical Field

[0001] This invention belongs to the field of environmental analytical chemistry and mass spectrometry, specifically relating to a targeted-non-targeted synergistic analysis method for aldehyde compounds based on stable isotope-labeled APMP-derived reagents and a dual-library system. Background Technology

[0002] Aldehydes are important polar oxygen-containing organic components in atmospheric particulate matter (PM2.5), playing a crucial role in the formation of secondary organic aerosols, particulate nucleation, and health effects. Due to the diversity of aldehydes, their varied structures, the low concentrations of some components, and their susceptibility to interference from complex matrices, accurate identification and quantitative analysis of aldehydes have always been challenging in the field of environmental analysis.

[0003] Existing aldehyde analysis methods mostly rely on derivatization combined with chromatography or chromatography-mass spectrometry detection, for example, using derivatization reagents to improve the response intensity and stability of the target analyte. However, traditional targeted analysis usually depends on available standards, and the range of compounds that can be quantified is limited, making it difficult to comprehensively cover the complex and diverse aldehyde components in actual PM2.5 samples.

[0004] Non-targeted analysis can broaden the scope of compound screening, but conventional non-targeted analysis mainly relies on precise mass numbers, secondary fragments, and matching with common spectral libraries, which suffers from problems such as misidentification, insufficient isomer differentiation, and low quantitative accuracy. In particular, when derivatization changes the structure of the target compound, the spectrum of the derivatized product often cannot be reliably matched with a general public spectral library, making it difficult to achieve both high-sensitivity detection by derivatization and high-coverage screening by non-targeted analysis.

[0005] Therefore, there is an urgent need to establish an analytical method that can organically combine derivatized targeted quantification with non-derivative non-targeted screening, so as to improve the identification coverage and reliability of aldehyde compounds in complex environmental samples while ensuring detection sensitivity and quantitative accuracy. Summary of the Invention

[0006] To overcome the shortcomings of existing technologies, the present invention aims to provide a targeted-non-targeted synergistic analysis method for aldehyde compounds based on stable isotope-labeled APMP derivatization reagents and a dual-library system. This method addresses the problems of limited coverage of targeted methods for aldehyde compounds, insufficient qualitative reliability of non-targeted methods, and difficulty in simultaneously achieving derivatization detection and matching with public spectral libraries in existing technologies. It enables synergistic screening and quantitative analysis of aldehyde compounds with and without standards.

[0007] The objective of this invention is achieved through the following technical solution:

[0008] A targeted-untargeted synergistic analysis method for aldehyde compounds based on stable isotope-labeled APMP-derived reagents and a dual-library system includes the following steps:

[0009] S1. Construct a dual-library system including derived spectral libraries and non-derived spectral libraries;

[0010] The derivatized spectral library is constructed by collecting chromatographic-mass spectrometry information after derivatizing aldehyde compounds with standards using APMP derivatizing reagent. The non-derivative spectral library is constructed by collecting data from underivative samples in IDA mode, screening them using a public mass spectrometry library, and verifying them by isotope derivatization using APMP derivatizing reagent.

[0011] S2. The samples to be tested are divided into a first sample and a second sample. The first sample is derivatized using APMP and then detected by liquid chromatography-mass spectrometry in MRM mode. The analysis is compared with the derivatized spectral library to perform targeted quantitative analysis of aldehyde compounds with standards. The second sample is not derivatized and is detected by liquid chromatography-high resolution mass spectrometry in IDA mode. The analysis is compared with the non-derivatized spectral library to perform non-targeted screening and confirmation of aldehyde compounds without standards.

[0012] Further, the APMP derivatizing reagent in step S1 includes d0-APMP and / or d3-APMP; wherein d0-APMP is 1-[4-(methylcarbamoyl)phenyl]-3-methyl-5-pyrazolone, and d3-APMP is 1-[4-(deuterated methylcarbamoyl)phenyl]-3-methyl-5-pyrazolone.

[0013] Furthermore, the construction of the derivatized spectral library in step S1 includes:

[0014] (1) The aldehyde compound with standard sample is derivatized with APMP derivatizing reagent to obtain the derivatized product;

[0015] (2) The retention time, parent ion, characteristic daughter ion, declustering voltage and collision energy parameters of the derivatives were collected by liquid chromatography-tandem mass spectrometry, and a derivative mass spectrometry library for targeted identification and quantitative analysis was established.

[0016] Furthermore, when constructing the derivatization library, the APMP derivatization reagent is d0-APMP.

[0017] Furthermore, the construction of the non-derived mass spectrometry library in step S1 includes:

[0018] (a) Liquid chromatography-high resolution mass spectrometry was used to acquire primary and secondary mass spectrometry information of the underrivatized sample using IDA mode;

[0019] (b) Match the primary and secondary mass spectrometry information with a public spectral library to obtain candidate aldehyde compounds;

[0020] (c) The candidate aldehyde compounds were verified by isotope derivatization using APMP derivatization reagent;

[0021] (d) Incorporate candidate aldehydes verified by isotope derivatization into the non-derivative mass spectrometry library.

[0022] Furthermore, the first-level scanning range of the IDA mode (information-dependent acquisition mode) described in step (a) is 50-1000 m / z, and the second-level scanning range is 50-1000 m / z.

[0023] Further, the public spectral library mentioned in step (b) includes one or more of NIST, MassBank and HMDB; the matching includes one or more of precise mass number matching, retention time matching and secondary mass spectrometry fragment matching.

[0024] Furthermore, the isotope-derived verification described in step (c) includes:

[0025] ① The candidate aldehyde compounds were derivatized using d0-APMP and d3-APMP, respectively;

[0026] ② The d0-APMP aldehyde derivatives and d3-APMP aldehyde derivatives obtained by detection;

[0027] ③ When the d0-APMP aldehyde derivative and the d3-APMP aldehyde derivative have corresponding mass shift characteristics, and the retention time and peak area response relationship between the two meet the preset requirements, the candidate aldehyde compound is confirmed to have passed the isotope derivatization verification.

[0028] Furthermore, the preset requirements are: consistent retention time (deviation ≤ 0.5 min) and peak area ratio of 0.5-2.0.

[0029] Furthermore, the relative standard deviation of the retention time of the d0-APMP aldehyde derivative and the d3-APMP aldehyde derivative in multiple repeated injections is less than 2%, and the deviation between the measured mass and the theoretical mass of the primary precursor ion is less than 5 ppm.

[0030] Furthermore, the APMP derivatization treatment in step S2 is carried out under alkaline conditions, with a derivatization temperature of 40–50 °C and a derivatization time of 10–20 min.

[0031] Further, the sample to be tested in step S2 is an extract of atmospheric particulate matter (PM2.5); the PM2.5 sample extract is prepared through the following steps:

[0032] The filter membrane containing PM2.5 was cut into pieces and added to an organic solvent for ultrasonic extraction; the resulting extract was filtered, concentrated, and reconstituted to obtain the sample to be tested.

[0033] Furthermore, the determination of the results of the collaborative analysis includes:

[0034] Aldehyde compounds that match the derivatization library are considered to be completely identified.

[0035] Aldehyde compounds that match the non-derived spectral library and are verified through isotope derivatization are considered to be essentially identified.

[0036] The present invention has the following advantages over the prior art:

[0037] 1. This invention improves the response intensity and detection sensitivity of aldehyde compounds in liquid chromatography-mass spectrometry by APMP derivatization, which is beneficial for the analysis of low-content aldehyde compounds in complex environmental samples.

[0038] 2. This invention establishes a derivatization spectral library for aldehyde compounds with standards, and can utilize multi-dimensional parameters such as retention time, parent ion, characteristic daughter ion, declustering voltage, and collision energy to achieve target identification and targeted quantification.

[0039] 3. This invention constructs a non-derivative spectral library for aldehydes for which no standards are available, and improves the reliability of non-targeted screening results and reduces the risk of false positives in non-targeted screening by using IDA acquisition, initial screening with public spectral libraries, and d0 / d3 isotope derivatization verification.

[0040] 4. This invention constructs and applies derivatized and non-derivatized spectral libraries in parallel, enabling synergistic screening and analysis of compounds with and without standards, thus improving the coverage of compounds. In addition, this invention has both targeted quantification and non-targeted screening capabilities, making it suitable for high-throughput analysis of trace aldehyde compounds in complex environmental samples such as PM2.5. Attached Figure Description

[0041] Figure 1 The 1H NMR spectrum of d0-APMP;

[0042] Figure 2 The carbon NMR spectrum of d0-APMP;

[0043] Figure 3 XIC diagram of the parent and daughter ions of the derivatized products of p-hydroxybenzaldehyde after d0-APMP derivatization;

[0044] Figure 4 This is a schematic diagram of the process for the targeted-non-targeted synergistic analysis method of aldehyde compounds based on APMP-derived reagents and a dual-library system according to the present invention. Detailed Implementation

[0045] The technical solution of the present invention will be further described below with reference to specific embodiments. However, those skilled in the art should understand that the following embodiments are only for illustrating the present invention and should not be regarded as limiting the present invention. Specific conditions not specified in the embodiments are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the reagents or instruments used are all conventional products obtained through commercial channels.

[0046] The ultra-high performance liquid chromatography of this invention uses an LC-30AD (Shimadzu Corporation, Japan) system and a 6500 Q-TRAP mass spectrometer (AB SCIEX, USA) equipped with an ion-driven turbine ESI source; the chromatographic column for ultra-high performance liquid chromatography is a Waters Atlantis T3 3μm (100mm × 2.1mm).

[0047] Example 1: Synthesis of d0-APMP

[0048] 436 mg of 1-(4-carboxyphenyl)-3-methyl-5-pyrazolone (CPMP, 0.2 mmol), 174 mg of methylaminohydrochloride (2.4 mmol), and 324 mg of 1-hydroxybenzotriazole (HOBTs, 0.2 mmol) were added to a flask. The mixture was placed in an ice bath at 0°C, and then 422 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 0.2 mmol) and 278 μL of triethylamine (TEA) were added. The mixture was reacted at room temperature for 18 h. The reaction solution was concentrated and purified by column chromatography (ethyl acetate: petroleum ether = 4:1). The solution was then evaporated to dryness to give 324 mg of d0-APMP, with a yield of 70%.

[0049] The above d0-APMP's 1H and 1C NMR spectra are as follows: Figure 1 , Figure 2 As shown, the structure was confirmed by mass spectrometry, hydrogen NMR, and carbon NMR as follows:

[0050] .

[0051] The amounts of CPMP, methylamine hydrochloride, HOBt, EDCI, and triethylamine can be adjusted proportionally according to the reaction scale.

[0052] Example 2: Synthesis of d3-APMP

[0053] This example is essentially the same as Example 1, except that the methylamine hydrochloride in Example 1 was replaced with deuterated methylamine hydrochloride (NH2-CD3·HCl, 180 mg, 2.4 mmol), while all other conditions remained the same, yielding 330 mg of d3-APMP in 70% yield. The structure was confirmed by mass spectrometry, 1H NMR, and 1C NMR as follows:

[0054] .

[0055] d3-APMP has the same or essentially the same reaction skeleton and chromatographic retention behavior as d0-APMP, and can be used for isotope derivatization verification or internal standard correction.

[0056] Example 3: Pretreatment of PM2.5 samples

[0057] Atmospheric particulate matter (PM2.5) samples were collected on quartz fiber filter membranes. Scissors were wiped with methanol to avoid sample contamination. One-quarter of the filter membrane was cut off, shredded, and placed in a 15 mL centrifuge tube. 10 mL of 80% (v / v) acetonitrile aqueous solution was added, and the mixture was extracted by sonication in an ice bath for 15 min. This process was repeated three times. The three extracts were combined, filtered through a 0.22 μm polytetrafluoroethylene (PTFE) filter membrane, and the filtrate was collected. The filtrate was concentrated to dryness by freezing and then reconstituted with acetonitrile to 1 mL to obtain the PM2.5 sample extract, which was then used for subsequent derivatization analysis or non-derivatized non-targeted analysis.

[0058] Example 4: Construction of Derivatized Spectral Libraries

[0059] Aldehyde compounds with standards were selected and derivatized with APMP derivatization reagent. The retention time (RT), parent ion (Q1), characteristic daughter ion (Q3), declustering voltage (DP), and collision energy parameter (CE) of the obtained aldehyde derivatives were collected by liquid chromatography-tandem mass spectrometry. Based on RT, Q1, Q3, DP, and CE, a derivatization spectral library was established.

[0060] In one specific embodiment, the aldehyde compound containing the standard includes one or more of the following: n-pentanal, n-hexanal, 4-hydroxybenzaldehyde, 3-hydroxybenzaldehyde, 4-hydroxy-3-methylbenzaldehyde, n-nonanal, vanillin, decanal, 3-allyl salicylaldehyde, undecanal, 4-hydroxy-3-methoxycinnamaldehyde, eugenol, peach aldehyde, and α-cyano-4-hydroxycinnamaldehyde. Acetonitrile is added to prepare aldehyde standard solutions with a concentration of 0.1 mmol / mL.

[0061] Take 50 μL of the above aldehyde standard solution, add 10 μL of d0-APMP derivatizing reagent (0.1 mmol / mL, prepared with acetonitrile) and 10 μL of 2% ammonia solution, and react at 45°C with shaking for 10 min. After the reaction, freeze and concentrate to remove the ammonia solution, add 500 μL of acetonitrile to redissolve, and perform LC-MRM analysis.

[0062] LC conditions: Waters Atlantis T3 column (100 mm × 2.1 mm, 3 μm); mobile phase A: 0.1% formic acid / water, mobile phase B: acetonitrile; gradient elution program: 0–15 min; flow rate: 0.3 mL / min; column temperature: 40°C; injection volume: 5 μL.

[0063] MS conditions: Electrospray positive ion mode was used, and the scanning method was multiple reaction monitoring (MRM); ion source temperature was 550°C; ion spraying voltage was 5500 V; atomizing gas (Gas1) was 30 psi; auxiliary gas (Gas2) was 15 psi; and curtain gas was 20 psi.

[0064] Since the chromatographic retention time and mass spectrometry fragmentation pattern of the derivatized products of d0-APMP and d3-APMP are exactly the same, this embodiment only uses d0-APMP as an example to establish a derivatization library. The obtained parameters such as retention time, parent ion, daughter ion and collision energy are also applicable to the internal standard labeled with d3-APMP.

[0065] The MRM parameters of 14 aldehyde compounds with standards are shown in Table 1 below; Figure 3 XIC diagram of the parent and daughter ions of the derivatized product after d0-APMP derivatization of p-hydroxybenzaldehyde.

[0066] Table 1

[0067] Serial Number Aldehydes Q1(m / z) Q3(m / z) RT(min) DP(V) CE(eV) 1 n-Pentanol 531 232 13.57 150 23 2 hexanol 545 232 14.82 150 25 3 p-Hydroxybenzaldehyde 567 232 17 160 27 4 3-Hydroxybenzaldehyde 581 232 19 160 26 5 4-Hydroxy-3-methylbenzaldehyde 587 232 18.24 145 30 6 nonanal 597 232 18.24 160 29 7 Vanillin 601 232 9 145 31 8 Decanal 607 232 10 155 31 9 3-Allyl salicylaldehyde 615 232 12.78 145 32 10 Undecaldehyde 623 232 8 160 33 11 4-Hydroxy-3-methoxycinnamon 627 232 16.35 165 45 12 Syringaldehyde 629 232 12.76 151 40 13 Peach aldehyde 634 232 16.02 165 35 14 α-Cyano-4-hydroxycinnamaldehyde 653 232 2.52 164 40

[0068] Example 5: Construction of non-derived spectral libraries

[0069] (a) Take the PM2.5 sample extract prepared in Example 3 or its quality control sample, and perform LC-high resolution mass spectrometry analysis directly in information-dependent acquisition (IDA) mode without derivatization to obtain the primary and secondary mass spectrometry information of the candidate compounds.

[0070] The IDA acquisition parameters are as follows: primary scan range m / z 50~1000, scan frequency 0.15 s / spectrum, declustering voltage 80V; intensity threshold for triggering secondary fragmentation >1000 cps, a maximum of 15 precursor ions can be triggered per cycle, dynamic exclusion time 10s; secondary collision energy 30 V±15 eV. Each sample is acquired 3 times.

[0071] (b) The acquired raw data in .Wiff format is converted to .mzML format suitable for non-targeted processing, and imported into MS-DIAL software for peak extraction, feature extraction, peak identification, and library matching to obtain candidate aldehyde compounds. Public spectral libraries may include one or more of NIST 20, MassBank, and HMDB. Matching metrics include one or more of the following: exact mass number, retention time, secondary mass spectrometry fragmentation information, and neutral loss features. The primary mass deviation is set to ≤0.01 Da, and the secondary mass deviation is set to ≤0.005 Da.

[0072] (c) For the candidate aldehyde compounds obtained from the initial screening, further isotopic derivatization verification was performed using d0-APMP and d3-APMP. Specifically, pure samples (if available) or enriched actual samples of the candidate aldehyde compounds were derivatized with d0-APMP and d3-APMP under the same conditions. The lightly labeled (d0) and heavily labeled (d3) derivatives were detected by MRM. If the retention times of the light and heavy derivatives were consistent (deviation ≤ 0.5 min) and the peak area ratio was within the range of 0.5-2, the candidate aldehyde compound was confirmed to be included in the non-derivative spectral library.

[0073] Example 6: Dual-library collaborative analysis method

[0074] Based on the foregoing embodiments, this invention provides a targeted-non-targeted synergistic analysis method for aldehyde compounds based on a dual-library system, as illustrated in the flowchart below. Figure 4 As shown, the specific process is as follows:

[0075] The PM2.5 sample extract prepared in Example 3 was divided into a first test sample and a second test sample:

[0076] First test sample: Take 50 μL of sample stock solution, add 10 μL of d0-APMP derivatizing reagent (0.1 mmol / mL, prepared with acetonitrile) and 10 μL of ammonia water, react at 45°C for 10 min, then add 10 μL of internal standard mixture labeled with d3-APMP derivatizing reagent (0.1 mmol / mL, prepared with acetonitrile), and detect according to the LC-MRM conditions of Example 4. Compare the detected chromatographic peaks with the derivatization library, and confirm the target analyte based on parameters such as retention time, MRM channel, and collision energy. Absolute quantification is performed using the isotope dilution method: calculate the peak area ratio of d0-APMP derivative to d3-APMP internal standard, and substitute it into the standard curve to calculate the concentration of aldehydes in the sample.

[0077] Second sample to be tested: Take 50 μL of sample stock solution, without derivatization, and directly perform LC-high resolution mass spectrometry analysis according to the IDA conditions in Example 6. The collected data are compared with the non-derivative spectral library for non-targeted screening and confirmation of aldehyde compounds without standards.

[0078] In one specific embodiment of the present invention, the results of the collaborative analysis are determined according to the following hierarchy:

[0079] (1) Aldehyde compounds that match the derivatization library and have standards and standard curves are identified as "completely determined" and their absolute concentrations are reported.

[0080] (2) Aldehyde compounds that match the non-derived spectral library and are verified by d0 / d3 isotope derivatization are determined to be "basically confirmed" and their relative concentrations are reported.

[0081] By using the above-mentioned result determination method, the present invention can distinguish and report the targeted quantitative results supported by the standard and the non-targeted screening results of compounds without the standard, thereby avoiding the direct use of the initial screening results of the public spectral library as the final result and improving the reliability of the aldehyde identification results in complex samples.

[0082] Example 7: Application Effect

[0083] The method described in this embodiment was used to analyze PM2.5 samples from a certain location. The samples were divided into two parts, and derivatized MRM analysis and non-derivatized IDA analysis were performed respectively using the method described above.

[0084] The results showed that a total of 55 aldehyde compounds were detected, as shown in Table 2. Among them, 14 aldehydes had commercially available standards and were completely identified by derivatization spectral library and absolutely quantified using the d3-APMP internal standard method, with concentration values ​​or ranges of 0.1-1000 ng / mL. The remaining 31 aldehydes did not have commercially available standards and were confirmed by initial screening using non-derivatization spectral library and verification by d0 / d3-APMP isotope derivatization. Although absolute quantification was not possible, the identification results were reliable and could be reported as "basically certain".

[0085] Method repeatability test: The same actual sample was analyzed 6 times under the same conditions. The relative standard deviation of the retention time of each compound was <2%, and the relative standard deviation of the peak area ratio of the light / heavy labeled derivatives was <15%, indicating that the method has good repeatability.

[0086] Table 2

[0087] Serial Number name Q1(m / z) Q3(m / z) RT(min) source 1 n-Pentanol 531 232 13.57 Completely certain 2 hexanol 545 232 14.82 Completely certain 3 p-Hydroxybenzaldehyde 567 232 17 Completely certain 4 3-Hydroxybenzaldehyde 581 232 19 Completely certain 5 4-Hydroxy-3-methylbenzaldehyde 587 232 18.24 Completely certain 6 nonanal 597 232 18.24 Completely certain 7 Vanillin 601 232 9 Completely certain 8 Decanal 607 232 10 Completely certain 9 3-Allyl salicylaldehyde 615 232 12.78 Completely certain 10 Undecaldehyde 623 232 8 Completely certain 11 4-Hydroxy-3-methoxycinnamon 627 232 16.35 Completely certain 12 Syringaldehyde 629 232 12.76 Completely certain 13 Peach aldehyde 634 232 16.02 Completely certain 14 α-Cyano-4-hydroxycinnamaldehyde 653 232 2.52 Completely certain 15 formaldehyde 475 232 7.89 Basically determined 16 Pentanal 531 232 12.34 Basically determined 17 furfural 541 232 2.1 Basically determined 18 Hexanal 545 232 7.8 Basically determined 19 benzaldehyde 545 232 13.69 Basically determined 20 heptanal 551 232 11 Basically determined 21 phenylacetaldehyde 559 232 14.77 Basically determined 22 4-Hydroxybenzaldehyde 565 232 2.1 Basically determined 23 Octal 573 232 15.76 Basically determined 24 2,4-Dimethylbenzaldehyde 579 232 13 Basically determined 25 4-Hydroxy-3-methylbenzaldehyde 581 232 2.1 Basically determined 26 3-Hydroxy-2-methylbenzaldehyde 581 232 13 Basically determined 27 Protocatechuic aldehyde 583 232 12.9 Basically determined 28 3-Chlorobenzaldehyde 586 232 2.1 Basically determined 29 Nononal 587 232 16.6 Basically determined 30 2-Methyl-3-phenyl-2-propenal 591 232 2.3 Basically determined 31 2-Methyl-3-phenyl-2-propenal-1 591 232 6.7 Basically determined 32 4-Propylbenzaldehyde 593 232 2.3 Basically determined 33 2-Ethyl-6-hydroxybenzaldehyde 595 232 2.1 Basically determined 34 Isonialdehyde 597 232 2.1 Basically determined 35 decanal 601 232 12 Basically determined 36 Naphthalene formaldehyde 601 232 17.3 Basically determined 37 2,3,4,5-Tetramethylbenzaldehyde 607 232 2.1 Basically determined 38 Veratral 607 232 9.6 Basically determined 39 Undeceneal 611 232 2.1 Basically determined 40 4-Ethynyl-1H-indole-3-carboxaldehyde 613 232 11 Basically determined 41 Undecaldehyde 614 232 12 Basically determined 42 2-Hydroxy-1-naphthaldehyde 615 232 18 Basically determined 43 2-(1-Benzofuran-5-yl)propionaldehyde 617 232 10.6 Basically determined 44 3-Methoxy-1-benzofuran-2-carboxaldehyde 619 232 2.1 Basically determined 45 Coniferaldehyde 621 232 2.1 Basically determined 46 Syringaldehyde 623 232 2.1 Basically determined 47 4-Undecyl lactone 629 232 18.8 Basically determined 48 1-Methoxy-2-naphthaldehyde-1 631 232 2.1 Basically determined 49 1-Methoxy-2-naphthaldehyde-2 631 232 11 Basically determined 50 1-Methoxy-2-naphthaldehyde-3 631 232 13.5 Basically determined 51 α-Cyano-4-hydroxycinnamic acid 634 232 2.1 Basically determined 52 trans-3,5-dimethoxy-4-hydroxycinnamaldehyde 653 232 2.1 Basically determined 53 3,5-Di-tert-butyl-4-hydroxybenzaldehyde-1 680 232 2.1 Basically determined 54 3,5-Di-tert-butyl-4-hydroxybenzaldehyde-2 680 232 6.7 Basically determined 55 (Z)-Hexadec-9-enal 684 232 2.1 Basically determined

[0088] Note: All compounds in this table were detected in actual PM2.5 samples. Among them, 14 aldehydes with commercially available standards have had their MRM parameters labeled in Example 3 and are classified as "fully definitive"; the remaining 41 aldehydes without standards but confirmed by dual-library co-validation are classified as "potentially definitive". Additionally, for aldehydes such as 1-methoxy-2-naphthaldehyde-1, 1-methoxy-2-naphthaldehyde-2, and 1-methoxy-2-naphthaldehyde-3, the suffixes 1, 2, and 3 in their names indicate that after derivatization, some MRM parameters showed multiple peaks, indicating the existence of isomers, and without subsequent standard supplementation, complete definitive determination is not possible.

[0089] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. Based on the present invention, some modifications or substitutions can be made, but these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of protection claimed by the present invention.

Claims

1. A targeted-non-targeted synergistic analysis method for aldehyde compounds based on stable isotope-labeled APMP-derived reagents and a dual-library system, characterized in that, Includes the following steps: S1. Construct a dual-library system including derived spectral libraries and non-derived spectral libraries; The derivatized spectral library is constructed by collecting chromatographic-mass spectrometry information after derivatizing aldehyde compounds with standards using APMP derivatizing reagent. The non-derivative spectral library is constructed by collecting data from underivative samples in IDA mode, screening them using a public mass spectrometry library, and verifying them by isotope derivatization using APMP derivatizing reagent. S2. The samples to be tested are divided into a first sample and a second sample. The first sample is derivatized using APMP and then detected by liquid chromatography-mass spectrometry in MRM mode. The analysis is compared with the derivatized spectral library to perform targeted quantitative analysis of aldehyde compounds with standards. The second sample is not derivatized and is detected by liquid chromatography-high resolution mass spectrometry in IDA mode. The analysis is compared with the non-derivatized spectral library to perform non-targeted screening and confirmation of aldehyde compounds without standards.

2. The collaborative analysis method according to claim 1, characterized in that, The APMP derivatizing reagents in step S1 include d0-APMP and / or d3-APMP; d0-APMP is 1-[4-(methylcarbamoyl)phenyl]-3-methyl-5-pyrazolone, and d3-APMP is 1-[4-(deuterated methylcarbamoyl)phenyl]-3-methyl-5-pyrazolone.

3. The collaborative analysis method according to claim 2, characterized in that, The construction of the derivatized spectral library in step S1 includes: (1) The aldehyde compound with standard sample is derivatized with APMP derivatizing reagent to obtain the derivatized product; (2) The retention time, parent ion, characteristic daughter ion, declustering voltage and collision energy parameters of the derivatives were collected by liquid chromatography-tandem mass spectrometry, and a derivative mass spectrometry library for targeted identification and quantitative analysis was established.

4. The collaborative analysis method according to claim 3, characterized in that, When constructing the derivatization library, the APMP derivatization reagent is d0-APMP.

5. The collaborative analysis method according to claim 2, characterized in that, The construction of the non-derived mass spectrometry library in step S1 includes: (a) Liquid chromatography-high resolution mass spectrometry was used to acquire primary and secondary mass spectrometry information of the underrivatized sample using IDA mode; (b) Match the primary and secondary mass spectrometry information with a public spectral library to obtain candidate aldehyde compounds; (c) The candidate aldehyde compounds were verified by isotope derivatization using APMP derivatization reagent; (d) Incorporate candidate aldehydes verified by isotope derivatization into the non-derivative mass spectrometry library.

6. The collaborative analysis method according to claim 5, characterized in that, The public spectral library mentioned in step (b) includes one or more of NIST, MassBank and HMDB; the matching includes one or more of precise mass number matching, retention time matching and secondary mass spectrometry fragment matching.

7. The collaborative analysis method according to claim 5, characterized in that, The isotope-derived verification described in step (c) includes: ① The candidate aldehyde compounds were derivatized using d0-APMP and d3-APMP, respectively; ② The d0-APMP aldehyde derivatives and d3-APMP aldehyde derivatives obtained by detection; ③ When the d0-APMP aldehyde derivative and the d3-APMP aldehyde derivative have corresponding mass shift characteristics, and the retention time and peak area response relationship between the two meet the preset requirements, the candidate aldehyde compound is confirmed to have passed the isotope derivatization verification.

8. The collaborative analysis method according to any one of claims 1 to 7, characterized in that, The APMP derivatization process described in step S2 is carried out under alkaline conditions, with a derivatization temperature of 40–50 °C and a derivatization time of 10–20 min.

9. The collaborative analysis method according to any one of claims 1 to 7, characterized in that, The sample to be tested in step S2 is an extract of atmospheric particulate matter (PM2.5); the PM2.5 sample extract is prepared through the following steps: The filter membrane containing PM2.5 was cut into pieces and added to an organic solvent for ultrasonic extraction; the resulting extract was filtered, concentrated, and reconstituted to obtain the sample to be tested.

10. The collaborative analysis method according to any one of claims 1 to 7, characterized in that, The determination of the results of the collaborative analysis includes: Aldehyde compounds that match the derivatization library are considered to be completely identified. Aldehyde compounds that match the non-derived spectral library and are verified through isotope derivatization are considered to be essentially identified.

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