Method for simultaneously detecting contents of 16 novel brominated flame retardants in human serum

Abstract

The invention provides a method for simultaneously detecting the content of 16 novel brominated flame retardants in human serum. The method comprises the following specific steps: S1, preparing a mixed standard working solution; s2, preparing an internal standard working solution; s3, drawing a standard working curve; s4, sample pretreatment: adding dichloromethane into a glass test tube, adding an internal standard working solution, carrying out water bath at 40 DEG C, carrying out nitrogen blowing until the solution is nearly dry, adding fetal calf serum and pure water, and carrying out vortex uniform mixing; the method comprises the following steps: absorbing a mixed solution of dichloromethane and acetonitrile and a sample to form an emulsion, carrying out vortex full extraction on the emulsion by using a dispersing agent and an extracting agent, and centrifuging at room temperature; finally, sucking an organic precipitation phase, filtering through a filter membrane, transferring into a small sample introduction bottle with a glass lining tube, and standing for on-machine detection; and S5, detection and result analysis: carrying out detection and result analysis on the content of the novel brominated flame retardant by adopting a gas chromatography-tandem triple quadrupole mass spectrometer. The method disclosed by the invention is simple in pretreatment, high in sensitivity and strong in practicability, and can be used for conventional biological monitoring of the NBFRs of the crowd.

Description

Technical Field

[0001] The invention belongs to the technical field of organic pollutant detection, and specifically relates to a method for simultaneously detecting the contents of 16 novel brominated flame retardants in human serum. Background Art

[0002] Traditional brominated flame retardants (BFRs) are widely used in plastics, textiles, adhesives, polyurethane foams, cable coatings, etc. due to their excellent flame retardant properties. Since BFRs are not chemically bound to the polymer matrix, they are leached into various environmental matrices, including air, sediment, water, and dust, during manufacturing, use, and disposal. A large number of studies have confirmed that BFRs may accumulate in organisms and cause significant adverse health effects on organisms, including interference with the thyroid endocrine system, neurotoxicity, immunotoxicity, genotoxicity, and reproductive toxicity. Due to their high persistence and toxicity, the Stockholm Convention listed pentabromodiphenyl ethers (BDEs) and octabromodiphenyl ether (Octa-BRE) as persistent organic pollutants (POPs) in 2009, followed by hexabromocyclododecane (HBCD) and decabromodiphenyl ether (BDE209) in 2013. As BFRs are gradually phased out, a series of new BFRs (NBFRs) have begun to enter the market and come into use.

[0003] Although described as an alternative to BFRs, NBFRs were first detected in the environment in the 1970s, and their use is expected to continue to increase. It is estimated that the annual production of NBFRs will exceed 1.3 million tons in 2020. China and the United States are the main producers of NBFRs, and the global production and consumption patterns are gradually shifting from BFRs to NBFRs. With the widespread use of NBFRs, they are constantly detected in various environmental media. On the other hand, NBFRs have the same semi-volatility and lipophilicity as polybrominated diphenyl ethers, and can be transported to different environmental media (dust, water, soil, sediment) worldwide through long-range atmospheric transport (LRAT) and accumulate in wildlife. Several studies have shown that NBFRs can affect the development of the nervous system, leading to impaired cognitive and motor functions, exacerbating anxious behaviors, attention problems, and intellectual disabilities, and also affect brain development by affecting thyroid hormone (TH) levels. Therefore, it is necessary to conduct research on the environmental exposure and population exposure characteristics of NBFRs, and the development of corresponding internal and external exposure detection methods is extremely necessary.

[0004] At present, some studies have been conducted on the detection methods of NBFRs in human blood. These analytical methods basically have limitations such as large amount of serum, high experimental cost (consumption of solid phase extraction columns and large amounts of organic solvents, etc.), cumbersome and time-consuming pretreatment process, and the detection items are concentrated on a few types of NBFRs. Therefore, based on the dispersed liquid-liquid extraction method, we used gas chromatography-tandem mass spectrometry to establish a detection method for NBFRs in human blood with a small amount of blood (200uL serum), a wide range of types (16 types), simple operation (pretreatment time 3 hours), and high sensitivity (0.020-0.750ng / mL). It can be effectively applied to large-scale epidemiological studies of the population and provide basic data for improving the exposure assessment of NBFRs in the population. Summary of the invention

[0005] The present invention provides a method for simultaneously detecting the contents of 16 novel brominated flame retardants in human serum. The results of the spiked recovery test show that the recovery range of the target NBFRs is 76.1%-120.5%, the relative standard deviation is between 0.60%-21.05%, and the method detection limit is between 0.020ng / mL and 0.750ng / mL. The method of the present invention has simple pretreatment, high sensitivity, and strong practicality, and can be used for conventional biological monitoring of NBFRs in the human population.

[0006] Reagent description of the present invention:

[0007] In the present invention, TBP is 2,4,6-tribromophenol, ATE is tribromophenyl allyl ether, TBX is 2,3,5,6-tetrabromo-p-xylene, PBT is pentabromotoluene, PBEB is pentabromophenyl ethane, DPTE is 2,3-dibromopropyl-2,4,6-tribromophenyl ether, HBB is hexabromobenzene, 2IPPDPP is 2-isopropylphenyl diphenyl phosphate, 4tBPDPP is p-tert-butylphenyl diphenyl phosphate, PBBA is propylene HCDBCO is hexachlorodibromooctane, TBB is 2-ethylhexyl-2,3,4,5-tetrabromobenzoate, TBBPA-BME is tetrabromobisphenol A dimethyl ether, DBPP is phenyl (di-tert-butylphenyl) phosphate, BTBPE is 1,2-bis (2,4,6-tribromophenoxy) ethane, TBPH is 3,4,5,6-tetrabromo-1,2-benzenedicarboxylic acid bis (2-ethylhexyl) ester, HBB-C 13 Hexabromobenzene is the isotopic internal standard.

[0008] In the present invention, the concentration of TBP standard is 100 μg / mL, the purity of ATE standard is 99.5%, the concentration of TBX standard is 100 μg / mL, the concentration of PBT standard is 1000 μg / mL, the concentration of PBEB standard is 100 μg / mL, the purity of DPTE standard is 99.5%, the concentration of HBB standard is 1000 μg / mL, the concentration of 2IPPDPP standard is 100 μg / mL, the purity of 4tBPDPP standard is 98.6%, the purity of PBBA standard is 98.7%, the concentration of HCDBCO standard is 100 μg / mL, the purity of TBB standard is 98.2%, the purity of TBBPA-BME standard is 98.5%, the concentration of DBPP standard is 100 μg / mL, the concentration of BTBPE standard is 100 μg / mL, and the purity of TBPH standard is 98.2%; the isotope HBB-C 13 The concentration was 10 μg / mL, and the above reagents were purchased from Tianjin Alta Company.

[0009] In the present invention, fetal bovine serum was purchased from Phygene, acetone, toluene, dichloromethane, n-hexane, acetonitrile and methanol were all chromatographically pure and purchased from Merck, Germany, deionized water was prepared by Milli-Q Academic ultrapure water instrument, and sulfuric acid (super pure) was purchased from Shanghai Lingfeng Reagent Company.

[0010] The technical solution adopted by the present invention to achieve the above object is: a method for simultaneously detecting the content of 16 new brominated flame retardants in human serum, characterized in that the specific steps are as follows:

[0011] S1: Prepare mixed standard working solution: accurately weigh appropriate amounts of TBP, ATE, TBX, PBT, PBEB, DPTE, HBB, 2IPPDPP, 4tBPDPP, PBBA, HCDBCO, TBB, TBBPA-BME, DBPP, BTBPE and TBPH standards, and prepare single standard solutions with a concentration of 100 μg / mL with n-hexane; pipette 100 μL of each single standard solution into a 100 mL volumetric flask; dilute to the mark with n-hexane to prepare a mixed standard working solution with a concentration of 100 ng / mL;

[0012] S2: Prepare the internal standard solution: draw 10 μg / mL of isotope internal standard HBB-C 13 Prepare an internal standard solution with a concentration of 100 ng / mL;

[0013] S3: Draw the standard working curve: Accurately pipette 1, 2, 4, 10, 20, 40, 100uL of the mixed standard solution into a glass test tube containing 200μL of dichloromethane, and add 20μL of the internal standard solution to each tube. After blowing with nitrogen until almost dry, add 200uL of fetal bovine serum and 4mL of pure water, and vortex for 30s to mix; Accurately pipette 200uL of the mixed solution of dichloromethane and 1mL of acetonitrile and quickly inject it into 200uL of the sample to form an emulsion, and use a dispersant and an extractant to extract the emulsion. After the turbid liquid was vortexed for 10 seconds for full extraction, it was centrifuged at 4700r / min for 5 minutes at room temperature; finally, the organic precipitate phase at the bottom was sucked out with a 1mL syringe, filtered through a 0.22μm nylon needle filter membrane and transferred to an injection vial with a glass liner tube for detection on the machine, and a series of working curves with concentrations of 0.5, 1, 2, 5, 10, 20, and 50ng / mL were obtained; finally, the standard working curve was drawn with the mass concentration ratio x of the compound and the internal standard to the peak area ratio y of the quantitative ion pair of the two;

[0014] S4: Sample pretreatment: Take 200μL of dichloromethane in a glass test tube, add 20μL of internal standard solution, place the glass test tube in a 40℃ water bath in a nitrogen blower, blow nitrogen until it is almost dry, add 200uL of fetal bovine serum and 4mL of pure water, and vortex for 30s to mix; accurately draw a mixture of 200uL of dichloromethane and 1mL of acetonitrile and quickly inject it into 200uL of sample to form an emulsion, use a dispersant and an extractant to vortex the emulsion for 10s to fully extract it, and then centrifuge it at 4700r / min at room temperature for 5min; finally, use a 1mL syringe to draw the organic precipitate phase at the bottom, filter it through a 0.22μm nylon needle filter membrane and transfer it to an injection vial with a glass liner tube for machine detection;

[0015] S5: Detection and result analysis: Gas chromatography-tandem triple quadrupole mass spectrometry was used to detect and analyze the content of new brominated flame retardants.

[0016] Furthermore, the novel brominated flame retardants include TBP, ATE, TBX, PBT, PBEB, DPTE, HBB, 2IPPDPP, 4tBPDPP, PBBA, HCDBCO, TBB, TBBPA-BME, DBPP, BTBPE and TBPH.

[0017] Furthermore, the internal standard concentration in the working curve in S3 is 10 ng / mL.

[0018] Furthermore, in S3 and S4, the dispersant is acetonitrile, the extractant is dichloromethane, and the volume ratio of the dispersant, the extractant and the emulsion is 25:1.

[0019] Furthermore, the volume ratio of the dispersant to the extractant is 5:1.

[0020] Furthermore, the gas chromatography-tandem triple quadrupole mass spectrometer in S5 is an Agilent 8890-7000D gas chromatography-tandem triple quadrupole mass spectrometer from the United States.

[0021] Furthermore, the gas chromatography conditions of the gas chromatography-tandem triple quadrupole mass spectrometer are:

[0022] The chromatographic column was an HP-5MS capillary column, the injection port temperature was 300°C, non-split injection was used, the column flow rate was 1.5 mL / min, the injection volume was 1.0 μL, and the optimized programmed temperature conditions were as follows: the initial temperature was 100°C and maintained for 3 min, then the temperature was increased to 200°C at 20°C / min and maintained for 1.5 min, then the temperature was increased to 250°C / min at 5°C / min, and then the temperature was increased to 280°C at 15°C / min and maintained for 4 min.

[0023] The method according to claim 5, characterized in that the specifications of the HP-5MS capillary chromatographic column are 15m×250um×0.25um.

[0024] Furthermore, the mass spectrometry conditions are:

[0025] The electron bombardment ion source mode was adopted, the ionization energy was 70eV, the ion source temperature was 280℃; the transfer line temperature was 300℃; the first and second quadrupole temperatures were both 150℃; the collision cell gases were nitrogen and helium, the nitrogen flow rate was 1.5mL / min, and the helium flow rate was 2.25ml / min; the solvent delay time was 4.00min; and the scanning mode was multiple reaction monitoring mode.

[0026] Compared with the prior art, the at least one technical solution adopted in the embodiments of this specification can achieve the following beneficial effects:

[0027] (1) The present invention uses a dichloromethane-acetonitrile dispersive liquid-liquid extraction method to extract 16 NBFRs in serum, and analyzes them by gas chromatography-tandem quadrupole mass spectrometry. A large number of optimization experiments are carried out on the chromatographic and mass spectrometric conditions and sample pretreatment methods from multiple aspects such as linearity, accuracy, precision, detection limit and quantification limit, so as to achieve the simultaneous detection and analysis of 16 NBFRs in serum;

[0028] (2) The results of the spiked recovery test of fetal bovine serum of the present invention showed that the recovery range of the target NBFRs was 76.1%-120.5%, the relative standard deviation was between 0.60%-21.05%, and the method detection limit was between 0.020ng / mL and 0.750ng / mL. This method has simple pretreatment, high sensitivity, and strong practicality, and can be used for routine biological monitoring of NBFRs in the human population. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on the drawings without paying creative work.

[0030] Figure 1 The total ion current of the mixed standard solution of 16 NBFRs is shown in Figure 1, where 1 is 2,4,6-tribromophenol, 2 is tribromophenyl allyl ether, 3 is 2,3,5,6-tetrabromo-p-xylene, 4 is pentabromotoluene, 5 is pentabromophenyl ethane, 6 is 2,3-dibromopropyl-2,4,6-tribromophenyl ether, 7 is hexabromobenzene and C 13 -hexabromobenzene, 8 is 2-isopropylphenyl diphenyl phosphate, 9 is p-tert-butylphenyl diphenyl phosphate, 10 is pentabromobenzyl acrylate, 11 is hexachlorodibromooctane, 12 is 2-ethylhexyl-2,3,4,5-tetrabromobenzoate, 13 is tetrabromobisphenol A dimethyl ether, 14 is phenyl (di-tert-butylphenyl) phosphate, 15 is 1,2-bis (2,4,6-tribromophenoxy) ethane, and 16 is 3,4,5,6-tetrabromo-1,2-benzenedicarboxylic acid bis (2-ethylhexyl) ester;

[0031] Figure 2 is the recovery rate of different combinations of extraction solvents;

[0032] Figure 3 Extraction efficiency with different dispersant volumes;

[0033] Figure 4 Extraction efficiency with different vortexing times. DETAILED DESCRIPTION

[0034] The embodiments of the present application are described in detail below with reference to the accompanying drawings.

[0035] The following describes the implementation methods of the present application through specific examples, and those skilled in the art can easily understand other advantages and effects of the present application from the contents disclosed in this specification. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. The present application can also be implemented or applied through other different specific implementation methods, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present application. It should be noted that, in the absence of conflict, the following embodiments and the features in the embodiments can be combined with each other. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in the field without making creative work belong to the scope of protection of the present application.

[0036] Example 1 Gas Chromatography Conditions and Mass Spectrometry Conditions

[0037] 1. Gas chromatography conditions

[0038] The chromatographic column was a HP-5MS capillary column (15m×250um×0.25um, Agilent, USA), the injection port temperature was 300°C, splitless injection was used, the column flow rate was 1.5mL / min, the injection volume was 1.0μL, and the optimized programmed temperature conditions were as follows: the initial temperature was 100°C for 3min, then increased to 200°C at 20°C / min and maintained for 1.5min, then increased to 250°C / min at 5°C / min, and then increased to 280°C at 15°C / min and maintained for 4min.

[0039] 2. Mass spectrometry conditions

[0040] The electron bombardment ion source mode was used, with an ionization energy of 70 eV, an ion source temperature of 280 °C, a transfer line temperature of 300 °C, a first and second quadrupole temperature of 150 °C, collision cell gas of nitrogen (flow rate of 1.5 mL / min) and helium (flow rate of 2.25 ml / min), and a solvent delay time of 4.00 min; scanning mode: multiple reaction monitoring mode (MRM), the dwell time was adjusted to 30 ms-100 ms according to the number of characteristic ion pairs in each time window to achieve 3-5 / s scanning cycles (Hz). The other mass spectrometry parameters are shown in Table 1.

[0041] Table 1. Retention times, ion pair parameters, collision energies, and electron multiplier gain parameters for 16 NBFRs and isotope internal standards

[0042]

[0043] Note: The ions marked with * are quantitative ion pairs.

[0044] Under the above conditions, the above 16 NBFRs mixed standard solutions were analyzed and determined. The results are as follows Figure 1 As shown in the figure, within 25.5 min, the separation effect of the mixed standard solution of 16 NBFRs was good and the peak shape was symmetrical.

[0045] Example 2 Optimization of sample pretreatment procedures

[0046] 1. Optimization of dispersant and extractant types

[0047] The present invention adds a standard solution (10 ng / mL) to fetal bovine serum and uses five dispersant and extractant combinations of n-hexane-acetone, toluene-methanol, toluene-acetone, dichloromethane-acetonitrile, and dichloromethane-methanol to perform dispersed liquid-liquid extraction. After extraction, the effects of different combinations on the recovery rate of NBFRs are compared on a machine (with the response of dichloromethane-acetonitrile as 100%).

[0048] Results Figure 2 , Figure 2 The results show that the recovery rate of most NBFRs using the combination of n-hexane-acetone and toluene-methanol is very low, the recovery rate of TBP and ATE using dichloromethane-methanol is not good, toluene-acetone and dichloromethane-acetonitrile have good recovery rates for each NBFRs. Considering that dichloromethane and acetonitrile are less toxic, and toluene and acetone are controlled explosive solvents, dichloromethane-acetonitrile was selected as the dispersant and extractant.

[0049] 2. Optimization of dispersant and extractant volumes

[0050] The volume of the extractant directly affects the enrichment multiple of the method. As the volume increases, the volume of the organic phase after centrifugation also increases, the concentration of the analyte after enrichment decreases, the enrichment multiple decreases, and the sensitivity also decreases. Therefore, in order to ensure a high enrichment multiple and sufficient organic phase for analysis, the volume of the extractant selected in the present invention is 200 μL.

[0051] The size of the dispersant volume directly affects the formation of the "water / dispersant / extractant / emulsion system", thereby affecting the extraction efficiency. The present invention uses 0.6, 0.8, 1.0, 1.2, and 1.4 mL of acetonitrile as the dispersant to investigate the extraction efficiency of the target compound with different dispersant volumes. The results are shown in Figure 3 When the volume of the dispersant increased to 1.0 mL, the extraction effect of most NBFRs reached its maximum value. There was no significant change in the extraction effect when the volume continued to increase. When the volume of the dispersant was large, the solubility of the analyte in water increased and it was not easy to be extracted, and the extraction efficiency would also decrease. After comprehensive consideration of the reagent dosage and extraction efficiency, 1.0 mL was selected as the dispersant dosage.

[0052] 3. Optimization of extraction time

[0053] The extraction time refers to the time from the injection of the extractant / dispersant system into the sample solution to the centrifugation. After the extraction solvent system enters the sample solution, the vortex method can be used to quickly form a large number of tiny droplets in the aqueous phase to increase the specific surface area, so that the target object can quickly migrate to the extraction solvent. The vortex oscillation time was set to 5, 10, 30, 60, and 120 seconds, respectively, and the recovery rates of 16 NBFRs were investigated. The results are shown in Figure 4The results showed that when the vortex oscillation time was between 10s and 30s, the extraction efficiency no longer showed an obvious increasing trend with the extraction time. When the vortex oscillation time was 60s, more protein precipitated in the sample, which was not conducive to subsequent liquid aspiration. Therefore, centrifugation could be performed directly after vortex oscillation for 10s.

[0054] Example 3 Method Verification

[0055] 1. Linear range, regression equation, detection limit and quantification limit of the working curve

[0056] In the linear range of 0.5-50 ng / mL, the regression equations of all NBFRs showed good linear relationships, R 2 Between 0.9974 and 0.999 8. The blank sample treatment solution was spiked at a concentration of 1.0 ng / mL, and the detection limit was 0.020-0.750 ng / mL when the signal-to-noise ratio was 3 and 10, respectively. See Table 2.

[0057] Table 2 Working curves, detection limits and quantification limits of 16 NBFRs in serum by gas chromatography-tandem mass spectrometry

[0058]

[0059] 2. Precision and accuracy of the method

[0060] Using blank serum as background, 16 NBFRs mixed standard solutions with low (1.0 ng / mL), medium (10 ng / mL), and high (20 ng / mL) concentrations were added for spike recovery test, and the results were shown in Table 3. The average recovery of this method ranged from 76.1% to 120.5%, with an RSD of 0.60%-21.05%.

[0061] Table 3 Results of spike recovery experiments of 16 NBFRs in serum by gas chromatography-tandem mass spectrometry (n=6)

[0062]

[0063] Note: “Low”, “Medium” and “High” correspond to concentrations of “2ng / mL”, “10ng / mL” and “30ng / mL” respectively; internal standard C 13 -HBB retention time is 13.24min.

[0064] Example 4 Determination of actual samples

[0065] The analytical method established in this study was used to analyze serum samples from 89 Shanghai residents, and the results are shown in Table 4. Twelve NBFRs were detected in the blood samples, with a detection rate of 2.27%-14.80%, with HBB (14.8%), TBB (10.2%), and TBBPA-BME (11.4%) having the highest detection rates, and the total NBFRs concentration ranged from 0 to 2.569 ng / mL.

[0066] Table 4 NBFRs test results of blood samples from residents of a community in Shanghai (n=89, ng / mL)

[0067]

[0068] From the above, it can be seen that the method of the present invention has simple pretreatment, high sensitivity, and strong practicability, and can be used for routine biological monitoring of NBFRs in the human population.

[0069] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable ordinary technicians in the field to understand the content of the present invention and implement it accordingly, and they cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made based on the essence of the content of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for simultaneously detecting the contents of 16 new brominated flame retardants in human serum, characterized in that: The specific steps are as follows: S1: Prepare mixed standard working solution: accurately weigh appropriate amounts of TBP, ATE, TBX, PBT, PBEB, DPTE, HBB, 2IPPDPP, 4tBPDPP, PBBA, HCDBCO, TBB, TBBPA-BME, DBPP, BTBPE and TBPH standards, and prepare single standard solutions with a concentration of 100 μg / mL with n-hexane; pipette 100 μL of each single standard solution into a 100 mL volumetric flask; dilute to the mark with n-hexane to prepare a mixed standard working solution with a concentration of 100 ng / mL; S2: Prepare the internal standard solution: draw 10 μg / mL of isotope internal standard HBB-C 13 Prepare an internal standard solution with a concentration of 100 ng / mL; S3: Draw the standard working curve: Accurately pipette 1, 2, 4, 10, 20, 40, 100uL of the mixed standard solution into a glass test tube containing 200μL of dichloromethane, and add 20μL of the internal standard solution to each tube. After blowing with nitrogen until almost dry, add 200uL of fetal bovine serum and 4mL of pure water, and vortex for 30s to mix; Accurately pipette 200uL of the mixed solution of dichloromethane and 1mL of acetonitrile and quickly inject it into 200uL of the sample to form an emulsion, and use a dispersant and an extractant to extract the emulsion. After the turbid liquid was vortexed for 10 seconds for full extraction, it was centrifuged at 4700r / min for 5 minutes at room temperature; finally, the organic precipitate phase at the bottom was sucked out with a 1mL syringe, filtered through a 0.22μm nylon needle filter membrane and transferred to an injection vial with a glass liner tube, and left for machine detection to obtain a series of working curves with concentrations of 0.5, 1, 2, 5, 10, 20, and 50ng / mL; finally, the standard working curve was drawn with the mass concentration ratio x of the compound and the internal standard to the peak area ratio y of the quantitative ion pair of the two; S4: Sample pretreatment: Take 200μL of dichloromethane in a glass test tube, add 20μL of internal standard solution, place the glass test tube in a 40℃ water bath in a nitrogen blower, blow nitrogen until it is almost dry, add 200uL of fetal bovine serum and 4mL of pure water, and vortex for 30s to mix; accurately draw a mixture of 200uL of dichloromethane and 1mL of acetonitrile and quickly inject it into 200uL of sample to form an emulsion, use a dispersant and an extractant to vortex the emulsion for 10s to fully extract it, and then centrifuge it at 4700r / min at room temperature for 5min; finally, use a 1mL syringe to draw the organic precipitate phase at the bottom, filter it through a 0.22μm nylon needle filter membrane and transfer it to an injection vial with a glass liner tube for machine detection; S5: Detection and result analysis: Gas chromatography-tandem triple quadrupole mass spectrometry was used to detect and analyze the content of new brominated flame retardants.

2. The method according to claim 1, characterized in that The novel brominated flame retardants include TBP, ATE, TBX, PBT, PBEB, DPTE, HBB, 2IPPDPP, 4tBPDPP, PBBA, HCDBCO, TBB, TBBPA-BME, DBPP, BTBPE and TBPH.

3. The method according to claim 1, characterized in that The internal standard concentration in the working curve of S3 is 10 ng / mL.

4. The method according to claim 1, characterized in that In the S3 and S4, the dispersant is acetonitrile, the extractant is dichloromethane, and the volume ratio of the dispersant, the extractant and the emulsion is 25:

1.

5. The method according to claim 4, characterized in that The volume ratio of the dispersant to the extractant is 5:

1.

6. The method according to claim 1, characterized in that The gas chromatography-tandem triple quadrupole mass spectrometer in S5 is an Agilent 8890-7000D gas chromatography-tandem triple quadrupole mass spectrometer from the United States.

7. The method according to claim 6, characterized in that The gas chromatography conditions of the gas chromatography-tandem triple quadrupole mass spectrometer are: The chromatographic column was an HP-5MS capillary column, the injection port temperature was 300°C, non-split injection was used, the column flow rate was 1.5 mL / min, the injection volume was 1.0 μL, and the optimized programmed temperature conditions were as follows: the initial temperature was 100°C and maintained for 3 min, then the temperature was increased to 200°C at 20°C / min and maintained for 1.5 min, then the temperature was increased to 250°C / min at 5°C / min, and then the temperature was increased to 280°C at 15°C / min and maintained for 4 min.

8. The method according to claim 7, characterized in that The specifications of the HP-5MS capillary chromatographic column are 15m×250um×0.25um.

9. The method according to claim 6, characterized in that The mass spectrometry conditions are: The electron bombardment ion source mode was adopted, the ionization energy was 70eV, the ion source temperature was 280℃; the transfer line temperature was 300℃; the first and second quadrupole temperatures were both 150℃; the collision cell gases were nitrogen and helium, the nitrogen flow rate was 1.5mL / min, and the helium flow rate was 2.25ml / min; the solvent delay time was 4.00min; and the scanning mode was multiple reaction monitoring mode.

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