Method for analyzing and identifying semi-volatile organic compounds in human serum sample

Abstract

The invention belongs to the technical field of substance content determination, and relates to a method for analyzing and identifying semi-volatile organic compounds in a human serum sample, which comprises the following steps: (1) sample pretreatment: removing impurities in a serum matrix through two times of liquid-liquid extraction, one time of solid-phase extraction and nitrogen blowing, and efficiently extracting and concentrating sample components to be analyzed; and (2) analyzing by using a gas chromatograph-mass spectrometer by using a dynamic multi-reaction monitoring (dMRM) method, and calculating the concentration of each to-be-detected target semi-volatile organic compound in the to-be-detected human serum sample according to an analysis result. According to the method, detail parameter conditions adopted in each step of the method are improved, so that a sensitive, accurate and scientific pretreatment and detection method can be provided for batch analysis and identification of various semi-volatile organic compounds in human serum samples.

Description

Technical Field

[0001] The present invention belongs to the technical field of substance content determination, and more specifically, relates to a method for analyzing and identifying semi-volatile organic compounds in human serum samples. Background Art

[0002] Semi-volatile organic compounds (SVOCs) generally refer to those with low volatility, insoluble in water but soluble in organic solvents, with boiling points between 170 and 350°C, and are likely to persist in the air, water, soil and other environments, with certain toxicity and bioaccumulation. Common ones include polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), phthalates (PAEs), etc. At present, there are many methods for detecting SVOCs in human blood at home and abroad, including liquid chromatography (LC), liquid / mass spectrometry (LC-MS), gas chromatography (GC), gas / mass spectrometry (GC-MS), gas chromatography-triple quadrupole tandem mass spectrometry (GC-MS / MS), etc. However, these methods can only measure a few substances with similar chemical properties or physical structures. However, in the real world, the human body is exposed to a variety of chemical substances, which may have cumulative effects and interactions on the human body.

[0003] In recent years, researchers have improved analytical methods to increase the number of SVOCs detected simultaneously. Sun Zhiwei et al. determined six novel brominated flame retardants (NBFRs) and eight polybrominated diphenylethers (PBDEs) in human serum based on GC-MS; Xia Yankai et al. determined 88 pesticides including organophosphorus (OPPs), OCPs, organic nitrogen, carbamates, pyrethroids (PYRs), and amide pesticides in 0.2 mL of human serum based on GC-MS / MS; Pulkrabova et al. determined 22 BFRs in 10 g of fish tissue homogenate based on GC-MS / MS; Varakina et al. determined 11 PCBs and 17 OCPs in 2 mL of human serum based on GC-MS / MS.

[0004] Since different SVOCs have different physical and chemical properties and structures, most studies have extracted a single or a few types of compounds in the sample, and few studies have conducted high-throughput and compatible simultaneous determination of multiple types of SVOCs in human serum samples. Therefore, establishing a scientific, accurate, and high-throughput analytical identification method for semi-volatile organic compounds in human serum samples has important application value for environmental exposure assessment in the fields of medicine and environmental science.

[0005] The research group of the inventor of the present invention has obtained a method for detecting semi-volatile organic compounds in blood through preliminary research (see Chinese patent document CN112379013A). Although it also discloses a method for detecting semi-volatile organic compounds in blood, which can detect 103 semi-volatile organic compounds (including 7 phthalates, 7 polychlorinated biphenyls, 16 polycyclic aromatic hydrocarbons, 5 polybrominated diphenyl ethers, and 68 pesticides) in human serum samples at the same time, the solid extraction and nitrogen blowing in the pre-treatment part of the experiment are manually operated, which is time-consuming and labor-intensive when multiple samples need to be processed, and is prone to human operational errors. Summary of the invention

[0006] In view of the above defects or improvement needs of the prior art, the object of the present invention is to provide a method for analyzing and identifying semi-volatile organic compounds in human serum samples, wherein by improving the detailed parameter conditions adopted in each step of the method, a sensitive, accurate and scientific pretreatment and detection method can be provided for batch analysis and identification of multiple semi-volatile organic compounds in human serum samples.

[0007] To achieve the above object, the present invention provides a method for analyzing and identifying semi-volatile organic compounds in human serum samples, comprising the following steps:

[0008] (1) Sample pretreatment, including the following sub-steps:

[0009] (1-1) Take 500 μL of the human serum sample to be tested, add ultrapure water to make the total volume 1 mL, then add 1 mL of acetonitrile, and after sealed ultrasonication for 10 mins, add 4 mL of n-hexane solvent; then, add 10 μL of the internal standard mixed solution with each internal standard substance concentration of 50 ppb prepared in advance, and then perform 1200 r / min oscillation extraction for 10 mins, 3000 r / min centrifugation for 10 mins, and then aspirate the supernatant, which is recorded as the first supernatant;

[0010] The internal standard mixture contains 24 internal standard substances, which are: phenanthrene-d10, dichlorvos-d6, 2,6-di-tert-butyl-p-cresol-d21, hexachlorobenzene-13d6, atrazine-d5, propargyl sulfoxide-d3, diazinon-d10, acetochlor-d11, polychlorinated biphenyl-28-d, polychlorinated biphenyl-52-d, polychlorinated biphenyl-101-d, polychlorinated biphenyl-118-d, polychlorinated biphenyl-138-d, polychlorinated biphenyl-153-d, polychlorinated biphenyl-180-d, malathion-d, chlorpyrifos-d10, fenthion-d6, ethion-d20, cypermethrin-d5, coumaphos-d, polybrominated diphenyl ether-77, perylene-d12, and 4,4'-DDT-13C12;

[0011] (1-2) 4 mL of a 1:1 mixed solvent of n-hexane / dichloromethane was added to the treated serum sample, followed by extraction at 1200 r / min for 10 mins and centrifugation at 3000 r / min for 10 mins, and then the supernatant was collected and recorded as the second supernatant;

[0012] (1-3) combining the first supernatant and the second supernatant, and drying them with nitrogen using a nitrogen blower; after drying, adding 2 mL of n-hexane to the mixture for re-dissolution, and vortexing for 30 seconds to obtain a reconstituted sample;

[0013] (1-4) placing the reconstituted sample in an automatic solid phase extractor for solid phase extraction, wherein the automatic solid phase extractor is operated according to the steps of needle washing - activation - air pushing - sample loading - rinsing - elution - air pushing - needle washing;

[0014] (1-5) After the solid phase extraction is completed, the sample is dried by a nitrogen blower in a 35°C water bath under the protection of pure nitrogen, and then 100 μL of n-hexane is added for re-dissolution, and vortexed for 30 seconds to obtain a concentrated liquid sample to be analyzed;

[0015] (2) analyzing the concentrated liquid sample to be analyzed obtained in step (1) by a dynamic multiple reaction monitoring (dMRM) method using a gas chromatograph-mass spectrometer, wherein the chromatographic conditions and mass spectrometric conditions of the gas chromatograph-mass spectrometer are previously optimized under a multiple reaction monitoring mode; the optimization under the multiple reaction monitoring mode is achieved by optimizing the detection sensitivity of each target semivolatile organic compound in the chromatographic and mass spectrometric detection process based on the preselected 106 types of target semivolatile organic compounds to be detected and based on the standard solution of each target semivolatile organic compound to be detected; and calculating the concentration of each target semivolatile organic compound in the human serum sample to be detected according to the analysis result;

[0016] The 106 target semivolatile organic compounds are 4-tert-butylphenol, 2,6-4-sec-butyl-di-tert-butylphenol, fluorene, 4-tert-octylphenol, ethoprophos, chlorpropham, trifluralin, fluazifop, cadusafos, phorate, α-hexachlorocyclohexane, hexachlorobenzene, simazine, β-hexachlorocyclohexane, atrazine, pentachloronitrobenzene, 3,5-di-tert-butyl-4-hydroxybenzyl alcohol, γ-hexachlorocyclohexane, olefins, pyruvate, pyroxene ... insectphos, propargyl sulfoxide, diazinon, phenanthrene, anthracene, methyl paraoxon, delta-hexachlor, ethidium, chlorpyrifos, methyl parathion, heptachlor, atrazine, promethazine, methyl pirimiphos, fenitrothion, PCB-28, PCB-52, PCB-101, PCB-118, PCB-138, PCB-153, PCB-18 0, Malathion, isopropyl metolachlor, chlorpyrifos, aldrin, fenthion, parathion, chlorpyrifos, pendimethalin, heptachlor B, chlorfenthion, isofenphos, fluoranthene, quinalphos, methidathion, o,p'-DD, chlorpyrifos, paclobutrazol, butachlor, pyrene, α-endosulfan, chlorpyrifos, chlorpyrifos, bromopropylphos, p,p'-DD, oxadiazine, dieldrin, endrin, o,p'-DD D, oxyfluorfen, herbicide, fluazifop-butyl, β-endosulfan, polybrominated diphenyl ether-28, polybrominated diphenyl ether-47, polybrominated diphenyl ether-99, polybrominated diphenyl ether-100, polybrominated diphenyl ether-153, polybrominated diphenyl ether-154, polybrominated diphenyl ether-183, p,p'-DDD, o,p'-DDT, ethion, triazole, p,p'-DDT, pyridazinphos-thiophene, phosmet, Benthion, benzo(a)anthracene, bifenthrin, cypermethrin, phosalone, cyhalothrin, permethrin, coumaphos, benzo(k)fluoranthene, benzo(b)fluoranthene, cyfluthrin, benzo(a)pyrene, cypermethrin, deltamethrin, benzo(g,h,i)perylene, dibenzo(a,h)anthracene, indeno(1,2,3-cd)pyrene;

[0017] The gas chromatography-mass spectrometer is specifically a gas chromatography-triple quadrupole tandem mass spectrometer.

[0018] As a further preferred embodiment of the present invention, in step (2), the chromatographic conditions are set as follows: using a DB-5MS capillary column, the temperature program is specifically as follows: initial temperature 45°C, heating to 130°C at 30°C / min, holding for 3mins, then heating to 180°C at 12°C / min, then heating to 240°C at 4°C / min, holding for 5mins, then heating to 300°C at 10°C / min, holding for 5mins; the injection port temperature is 280°C, the carrier gas is high-purity helium with a purity of not less than 99.999%, the carrier gas flow rate is 1mL / min, non-split injection, and the injection volume is 1μL;

[0019] The mass spectrometry conditions were set as follows: electron impact (EI) ion source, ionization energy 70 eV, ion source temperature 280°C, transfer line temperature 280°C, and solvent delay 5.5 mins.

[0020] As a further preferred embodiment of the present invention, in step (2), the ion pairs and collision energies used in gas chromatography-triple quadrupole tandem mass spectrometry are shown in the following table:

[0021]

[0022]

[0023]

[0024] As a further preference of the present invention, in sub-step (1-5), the sample is blown dry in a 35°C water bath under the protection of pure nitrogen, specifically: the nitrogen blower parameters are set to a pressure of 0.1 MPa, an initial needle height of 0 mm, and a needle speed of 1 mm / min, and the sample is blown dry in a 35°C water bath under the protection of pure nitrogen.

[0025] As a further preferred embodiment of the present invention, sub-steps (1-3), (1-4) and (1-5) are performed on multiple sample batches.

[0026] Through the above technical scheme conceived by the present invention, compared with the prior art, the analysis and identification method of semi-volatile organic compounds in human serum samples of the present invention can simultaneously detect 106 SVOCs in human serum, including PCBs (7 species), PBDEs (7 species), PAHs (13 species), synthetic phenolic antioxidants (SPAs, 4 species), OPPs (30 species), OCPs (19 species), PYRs (7 species), herbicides (19 species) and 24 corresponding internal standard substances. The method of the present invention is firstly a pre-treatment method of 2 liquid-liquid extractions, 1 solid phase extraction, and nitrogen blowing, which can better remove impurities in serum and extract target analytes, and then use GC-MS / MS to accurately measure 106 semi-volatile organic compounds and 24 internal standards in human serum in batches, saving detection time (as exemplified in the following embodiments, the 106 compounds measured can all be peaked within 42 minutes, completing GC-MS / MS detection), and improving sample utilization.

[0027] The present invention can preferably use specially set ion pairs and collision energies for GC-MS / MS detection, and can accurately perform qualitative and quantitative analysis on 106 target analytes, saving detection time and improving sample utilization.

[0028] The method for analyzing and identifying semi-volatile organic compounds in human serum samples of the present invention, on the one hand, removes impurities from serum samples by two liquid-liquid extractions, one solid phase extraction, and nitrogen blowing in pretreatment, and extracts and concentrates the analytes; on the other hand, as illustrated in the embodiments below, the dMRM method established by GC-MS / MS can achieve one-shot injection (1 μL, 42 mins) to simultaneously detect 106 SVOCs and 24 internal standards, thereby reducing time costs and improving sample utilization.

[0029] The present invention can detect 106 SVOCs at the same time, and 4 SPAs are newly added compared to the 103 SVOCs detected by the previous patent CN112379013A. SPAs have caused ubiquitous pollution in indoor and outdoor environments around the world. Multiple in vivo and in vitro studies have reported the toxic effects of various genotoxicity, hepatotoxicity, endocrine disruption and carcinogenicity of SPAs. The present invention can simultaneously perform qualitative and quantitative determination of 106 SVOCs including SPAs in serum in batches. Moreover, compared to the manual operation and manual nitrogen blowing operation of Florisil solid phase extraction cartridges used in the previous patent CN112379013A, which are time-consuming and cannot avoid manual operation errors, the present invention adopts an automated solid phase extractor and nitrogen blowing instrument, which can simultaneously complete the processing of multiple samples in batches, saving time, reducing manual operation errors, and reducing experimental error rates, and is suitable for material determination of large-scale population samples.

[0030] The method of the present invention can realize the qualitative and quantitative analysis of a variety of semi-volatile organic compounds in human serum samples in batches in a short period of time, saving detection time and improving sample utilization. It can be applied to scenarios such as collecting regional environmental big data to determine the exposure degree of various semi-volatile organic compounds in human blood, and provides technical support for the evaluation of environmental pollutant exposure levels in medical and environmental science research. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 It is a linear fitting curve of detecting Dichlorvos by the method of the embodiment of the present invention.

[0032] Figure 2 It is a linear fitting curve for detecting δ-hexachloride (δ-HCH) by the method of the embodiment of the present invention. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0034] In general, the method for analyzing and identifying semi-volatile organic compounds in human serum samples of the present invention comprises the following steps: (1) sample pretreatment to obtain sample components to be analyzed; (2) on-machine detection: the concentrated sample components are loaded into a sample injection bottle and detected under a dynamic multiple reaction monitoring (dMRM) method established by GC-MS / MS. That is: (1) human serum sample pretreatment, through two liquid-liquid extractions, one solid phase extraction, and nitrogen blowing, impurities in the serum matrix are removed, and the sample components to be analyzed are efficiently extracted and concentrated; (2) on-machine detection, 106 semi-volatile organic compounds are batch-determined within 42 minutes using gas chromatography-triple quadrupole tandem mass spectrometry (GC-MS / MS).

[0035] The GC-MS / MS used in the examples below is gas chromatography (Agilent 8890GC)-triple quadrupole mass spectrometry (Agilent 7010B MS).

[0036] Example

[0037] A method for determining SVOCs in serum samples using two liquid-liquid extractions-one solid phase extraction and GC-MS / MS, the operating steps are as follows:

[0038] (1) Sample (human serum) pretreatment, and then obtain the sample to be analyzed:

[0039] Take 500 μL of human serum sample and add it to a 10 mL digestion tube, add ultrapure water to make the total volume 1 mL, then add 1 mL of acetonitrile, screw on the lid and seal it for 10 mins, then add 4 mL of n-hexane solvent. Add 10 μL of the pre-prepared 50 ppb internal standard solution (i.e., 10 μL of internal standard mixed solution containing 24 internal standard substances, each with a concentration of 50 ppb) to each sample, screw on the lid, perform 1200 r / min oscillation extraction for 10 mins, centrifuge at 3000 r / min for 10 mins, and then use a glass dropper to aspirate the supernatant (about 4 mL) in the digestion tube and transfer it to a round-bottom glass tube. Continue to add 4 mL of a 1:1 volume ratio of n-hexane / dichloromethane mixed solvent to the original digestion tube, repeat the above oscillation and centrifugation operations, and use a glass dropper to aspirate the supernatant (4 mL to 5 mL) and combine it with the first supernatant. After merging, transfer the round-bottom glass tube containing the supernatant to a nitrogen blower. Set the nitrogen blower parameters to 0.1 MPa pressure, 0 mm initial needle height, and 1 mm / min needle speed. Blow dry the sample under the protection of pure nitrogen. After drying, use a bottle-mouth dispenser to add 2 mL of n-hexane to the round-bottom glass tube for re-dissolution and vortex for 30 seconds.

[0040] After re-dissolution, the samples are placed in an automatic solid phase extractor for solid phase extraction. The automatic solid phase extractor can be used to perform solid phase extraction on multiple parallel samples at the same time. Moreover, if the number of samples exceeds the number of samples that can be processed by the automatic solid phase extractor at a single time, the next row of samples can be processed in a cycle. The parameters of the automatic solid phase extractor are as follows: (1) needle washing, the waste liquid is discharged; (2) activation (i.e., first use 3 mL of a mixed solution of n-hexane / dichloromethane with a volume ratio of 8:2 for activation treatment, and then use 3 mL of a mixed solution of n-hexane / dichloromethane with a volume ratio of 1:1 for activation treatment), and the waste liquid is discharged; (3) air pushing, the waste liquid is discharged; (4) sample loading: 2 mL, collection; (5) rinse: 1 mL, collection; (6) elution: 6.4 mL, collection (during elution, specifically, first use 2.2 mL of a mixed solution of n-hexane / dichloromethane with a volume ratio of 8:2, then use 2 mL of a mixed solution of n-hexane / ethyl acetate with a volume ratio of 8:2, and finally use 2.2 mL of a mixed solution of n-hexane / dichloromethane with a volume ratio of 1:1); (7) air pushing: collection; (8) needle washing, the waste liquid is discharged; (9) end or start the next cycle (i.e., process the next row of samples in the solid phase extractor).

[0041] After the solid extraction is completed, continue to blow dry the sample in a 35°C water bath under the protection of pure nitrogen, add 100 μL of n-hexane to dissolve it, obtain the final concentrated solution to be analyzed, and vortex for 30 seconds. Transfer the concentrated solution to the inner tube, put it into a 2mL sample bottle, cover the bottle cap, and wait for analysis on the machine.

[0042] (2) The obtained sample components were placed in a sample injection bottle, and the created dynamic multiple reaction monitoring (dMRM) method was called up in the GC-MS / MS to measure the sample:

[0043] The setting conditions of the GC-MS / MS chromatography and mass spectrometry are:

[0044] Chromatographic conditions: DB-5MS capillary column (30m×0.25mm×0.25μm); temperature program: initial temperature 45°C, heated to 130°C at 30°C / min, maintained for 3min, then heated to 180°C at 12°C / min, then heated to 240°C at 4°C / min, maintained for 5min, then heated to 300°C at 10°C / min, maintained for 5min; injection port temperature 280°C, carrier gas high-purity helium (99.999%), carrier gas flow rate 1mL / min, non-split injection, injection volume 1μL;

[0045] Mass spectrometry conditions: electron impact (EI) ion source, ionization energy 70 eV, ion source temperature 280 °C, transfer line temperature 280 °C, solvent delay 5.5 mins.

[0046] The dynamic multiple reaction monitoring (dMRM) method for 106 semi-volatile organic compounds was established as follows: a mixed standard of 106 semi-volatile organic compounds and 24 internal standards with a concentration of 100 ppm were injected separately, the samples were scanned in full scan mode, and the raw data obtained in GC-MS / MS were imported into the Unknown Analysis software and the MassHunter Qualitative Analysis software for deconvolution and identification of chromatographic peaks. 20 standard mass spectrometry libraries were used for matching and analysis to obtain accurate qualitative results; then, the mass-to-charge ratio of the specific parent ion was determined in the qualitative analysis software, the time and scan type (product ion) were set in the ion scan (SIM) mode, the precursor ion was set in the scan segment according to the mass-to-charge ratio of the specific parent ion determined in the previous step, the MS2 scan endpoint mass was rounded according to the precursor ion, and the daughter ion fragments with higher responsiveness were determined under the conditions of CE (eV) of 10 and 20; finally, the dMRM mode was used to scan and optimize the collision energy, and the elution time, two pairs of optimal parent ion / daughter ion pairs, and collision energy of each semi-volatile organic compound were determined one by one. The optimization results are shown in Table 1. The optimization results can be saved, for example, set in GC-MS / MS, and the method is called out each time the sample is measured, and the sample to be tested is analyzed and measured by GC-MS / MS according to the conditions set by the above chromatography and mass spectrometry.

[0047] According to the above chromatographic and mass spectrometric conditions, the dMRM method in Table 1 was used for detection. This method was used to batch determine 106 SVOCs (7 PCBs, 7 PBDEs, 13 PAHs, 4 SPAs, 30 OPPs, 19 OCPs, 7 PYRs, 19 herbicides) and 24 internal standards (ISTD).

[0048] In the process of research and development, the present invention actually attempted to determine 129 kinds of semi-volatile organic compounds, including amphetamine, cyanazine, metamitron, fluazifop, butyl benzyl phthalate, di-n-butyl phthalate, di-(2-ethylhexyl) phthalate, diethyl phthalate, diisobutyl phthalate, dimethyl phthalate, di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1,2-di-1 There are 23 semivolatile organic compounds, including dioctyl ester, naphthalene, acenaphthylene, acenaphthene, dichlorvos, cypermethrin, 2,6-di-tert-butylphenol, 2,6-di-tert-butyl-4-hydroxy-4-methyl-2,5-cyclohexanedione, 2,6-di-tert-butylbenzoquinone, 2,4-di-tert-butylphenol, 2,6-di-tert-butyl-p-cresol, and 2,4,6-tri-tert-butylphenol, but their standard curves have poor linearity (R 2 <0.999), it cannot be accurately quantified, so it is no longer included in the test. Taking Dichlorvos as an example, the linearity of its standard curve (such as Figure 1As shown, R 2 =0.1768, greater than 0.999). The 106 SVOCs retained by the method of the present invention have good compatibility during determination, and all of them can show peaks on the chromatogram and the standard curve has good linearity, so that various SVOCs can be accurately analyzed qualitatively and quantitatively. Taking δ-hexachloride (δ-HCH) as an example, Figure 2 As shown, the standard curve has good linearity.

[0049] Table 1: 106 SVOCs and 24 internal standards detected based on the dMRM method

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056] According to the above method, 100 serum samples were automatically processed in batches using a nitrogen blowdown apparatus and a solid phase extractor; the detection rates of 106 SVOCs in these 100 serum samples are shown in Table 2 (detection rate = number of samples above the detection limit ÷ total number of samples × 100%).

[0057] Table 2: Detection rate (%) of 106 SVOCs in 100 serum samples

[0058]

[0059]

[0060] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for analyzing and identifying semi-volatile organic compounds in human serum samples, characterized in that: The following steps are involved: (1) Sample pretreatment, including the following sub-steps: (1-1) Take 500 μL of the human serum sample to be tested, add ultrapure water to make the total volume 1 mL, then add 1 mL of acetonitrile, and after sealed ultrasonication for 10 mins, add 4 mL of n-hexane solvent; then, add 10 μL of the internal standard mixed solution with each internal standard substance concentration of 50 ppb prepared in advance, and then perform 1200 r / min oscillation extraction for 10 mins, 3000 r / min centrifugation for 10 mins, and then aspirate the supernatant, which is recorded as the first supernatant; The internal standard mixture contains 24 internal standard substances, which are: phenanthrene-d10, dichlorvos-d6, 2,6-di-tert-butyl-p-cresol-d21, hexachlorobenzene-13d6, atrazine-d5, propargyl sulfoxide-d3, diazinon-d10, acetochlor-d11, polychlorinated biphenyl-28-d, polychlorinated biphenyl-52-d, polychlorinated biphenyl-101-d, polychlorinated biphenyl-118-d, polychlorinated biphenyl-138-d, polychlorinated biphenyl-153-d, polychlorinated biphenyl-180-d, malathion-d, chlorpyrifos-d10, fenthion-d6, ethion-d20, cypermethrin-d5, coumaphos-d, polybrominated diphenyl ether-77, perylene-d12, and 4,4'-DDT-13C12; (1-2) 4 mL of a 1:1 mixed solvent of n-hexane / dichloromethane was added to the treated serum sample, followed by extraction at 1200 r / min for 10 mins and centrifugation at 3000 r / min for 10 mins, and then the supernatant was collected and recorded as the second supernatant; (1-3) combining the first supernatant and the second supernatant, and drying them with nitrogen using a nitrogen blower; after drying, adding 2 mL of n-hexane to the mixture for re-dissolution, and vortexing for 30 seconds to obtain a reconstituted sample; (1-4) placing the reconstituted sample in an automatic solid phase extractor for solid phase extraction, wherein the automatic solid phase extractor is operated according to the steps of needle washing - activation - air pushing - sample loading - rinsing - elution - air pushing - needle washing; (1-5) After the solid phase extraction is completed, the sample is dried by a nitrogen blower in a 35°C water bath under the protection of pure nitrogen, and then 100 μL of n-hexane is added for re-dissolution, and vortexed for 30 seconds to obtain a concentrated liquid sample to be analyzed; (2) analyzing the concentrated liquid sample to be analyzed obtained in step (1) by a dynamic multiple reaction monitoring (dMRM) method using a gas chromatograph-mass spectrometer, wherein the chromatographic conditions and mass spectrometric conditions of the gas chromatograph-mass spectrometer are previously optimized under a multiple reaction monitoring mode; the optimization under the multiple reaction monitoring mode is achieved by optimizing the detection sensitivity of each target semivolatile organic compound in the chromatographic and mass spectrometric detection process based on the preselected 106 types of target semivolatile organic compounds to be detected and based on the standard solution of each target semivolatile organic compound to be detected; and calculating the concentration of each target semivolatile organic compound in the human serum sample to be detected according to the analysis result; The 106 target semivolatile organic compounds are 4-tert-butylphenol, 2,6-4-sec-butyl-di-tert-butylphenol, fluorene, 4-tert-octylphenol, ethoprophos, chlorpropham, trifluralin, fluazifop, cadusafos, phorate, α-hexachlorocyclohexane, hexachlorobenzene, simazine, β-hexachlorocyclohexane, atrazine, pentachloronitrobenzene, 3,5-di-tert-butyl-4-hydroxybenzyl alcohol, γ-hexachlorocyclohexane, olefins, pyruvate, pyroxene ... insectphos, propargyl sulfoxide, diazinon, phenanthrene, anthracene, methyl paraoxon, delta-hexachlor, ethidium, chlorpyrifos, methyl parathion, heptachlor, atrazine, promethazine, methyl pirimiphos, fenitrothion, PCB-28, PCB-52, PCB-101, PCB-118, PCB-138, PCB-153, PCB-18 0, Malathion, isopropyl metolachlor, chlorpyrifos, aldrin, fenthion, parathion, chlorpyrifos, pendimethalin, heptachlor B, chlorfenthion, isofenphos, fluoranthene, quinalphos, methidathion, o,p'-DD, chlorpyrifos, paclobutrazol, butachlor, pyrene, α-endosulfan, chlorpyrifos, chlorpyrifos, bromopropylphos, p,p'-DD, oxadiazine, dieldrin, endrin, o,p'-DD D, oxyfluorfen, herbicide, fluazifop-butyl, β-endosulfan, polybrominated diphenyl ether-28, polybrominated diphenyl ether-47, polybrominated diphenyl ether-99, polybrominated diphenyl ether-100, polybrominated diphenyl ether-153, polybrominated diphenyl ether-154, polybrominated diphenyl ether-183, p,p'-DDD, o,p'-DDT, ethion, triazole, p,p'-DDT, pyridazinphos-thiophene, phosmet, Benthion, benzo(a)anthracene, bifenthrin, cypermethrin, phosalone, cyhalothrin, permethrin, coumaphos, benzo(k)fluoranthene, benzo(b)fluoranthene, cyfluthrin, benzo(a)pyrene, cypermethrin, deltamethrin, benzo(g,h,i)perylene, dibenzo(a,h)anthracene, indeno(1,2,3-cd)pyrene; The gas chromatography-mass spectrometer is specifically a gas chromatography-triple quadrupole tandem mass spectrometer.

2. The method for analyzing and identifying semi-volatile organic compounds in human serum samples according to claim 1, characterized in that: In step (2), the chromatographic conditions are set as follows: using a DB-5MS capillary column, the temperature program is specifically as follows: initial temperature 45°C, heating to 130°C at 30°C / min, holding for 3mins, then heating to 180°C at 12°C / min, then heating to 240°C at 4°C / min, holding for 5mins, then heating to 300°C at 10°C / min, holding for 5mins; the injection port temperature is 280°C, the carrier gas is high-purity helium with a purity of not less than 99.999%, the carrier gas flow rate is 1mL / min, non-split injection, and the injection volume is 1μL; The mass spectrometry conditions were set as follows: electron impact (EI) ion source, ionization energy 70 eV, ion source temperature 280°C, transfer line temperature 280°C, and solvent delay 5.5 mins.

3. The method for analyzing and identifying semi-volatile organic compounds in human serum samples according to claim 1, characterized in that: In step (2), the ion pairs and collision energies used by gas chromatography-triple quadrupole tandem mass spectrometry are shown in the following table:

4. The method for analyzing and identifying semi-volatile organic compounds in human serum samples according to claim 1, characterized in that: In sub-step (1-5), the sample is blown dry in a 35°C water bath under the protection of pure nitrogen. Specifically, the nitrogen blower parameters are set to pressure 0.1 MPa, initial needle height 0 mm, needle speed 1 mm / min, and the sample is blown dry in a 35°C water bath under the protection of pure nitrogen.

5. The method for analyzing and identifying semi-volatile organic compounds in human serum samples according to claim 1, characterized in that: Sub-steps (1-3), (1-4) and (1-5) are performed on multiple sample batches.

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