Human body non-invasive biological matrix PFAS recognition system and synchronous detection method

Abstract

The invention relates to a human body non-invasive biological matrix PFAS identification system and a synchronous detection method. The system establishment method comprises the following steps: S1, screening key enterprise, industry and regional characteristic pollutants PFAS; s2, screening the PFAS based on ecological risks, human health risks and PMT / PBT material characteristics of the PFAS; s3, establishing a biological detection list based on the accumulation characteristics of the PFAS in various non-invasive biological matrixes; and S4, carrying out PFAS biological monitoring, and prompting the PFAS which needs to be preferentially controlled according to the human body biological monitoring condition of the PFAS. According to the technical scheme, non-invasive biological monitoring is carried out by screening characteristic PFAS, the method is clear in target, efficient, low in cost and suitable for large-sample population investigation, and the constructed non-invasive matrix (hair, fingernails and urine) PFAS synchronous detection method is simple in pretreatment process, convenient to operate, low in coexisting substance interference and stable and efficient in recovery rate.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of new pollutant detection of biological matrix, and particularly relates to a human non-invasive biological matrix PFAS identification system and synchronous detection method. BACKGROUND

[0002] Per-and polyfluoroalkyl substances (PFAS) are a large class of synthetic chemicals, as a new type of persistent organic pollutants, PFAS is difficult to degrade in the environment and easy to accumulate in the body. Therefore, it is extremely important to carry out biological monitoring of human PFAS for assessing exposure level and effectively preventing health risks. At present, the biological monitoring matrix of human PFAS includes blood, hair, nails, urine, placenta and breast milk, etc. There are certain differences in PFAS research among different matrices. Blood samples are difficult to obtain the cooperation of sampling objects and sampling legality due to a certain degree of damage to the human body by sampling method, which limits the application of this human biological sample to a certain extent; placenta and breast milk samples have great limitations in the scope of application of the population because they are aimed at specific population; while urine, hair and nail samples are easy to obtain, and can be sampled in large volume, which is easy to detect and has good application prospect in large-scale population investigation. Therefore, it is of great significance to carry out the identification system and multiple synchronous detection methods of non-invasive biological matrix PFAS for understanding the pollution status of PFAS and its impact on human health.

[0003] PFAS has a wide variety of species, different physicochemical properties, complex environmental migration and transformation paths, and significant differences in accumulation characteristics and toxic effects in organisms. Therefore, in the identification process, how to accurately screen out target PFAS compounds with significant potential health risks based on the characteristics of specific pollution sources (such as industrial point sources, migration and transformation by-products, and consumer product releases) and prioritize human biological monitoring becomes a key technology. The detection of PFAS in biological samples is divided into two parts: sample pretreatment and instrument detection and analysis. Due to the different contents of PFAS in different human biological samples, how to perform scientific pretreatment to reduce the influence of matrix effect on the results and obtain lower detection limit and better recovery rate is the technical core of the pretreatment method. Currently, the pretreatment method for PFAS in three types of non-invasive biological matrixes such as urine, nails and hair is mainly solid phase extraction, but the sampling amount, sample pretreatment, selection of solid phase column, extraction solvent and process all have a great influence on the recovery rate. In terms of instrument analysis, since the content of PFAS in human biological matrix is usually at trace or ultra-trace level, the detection method often needs to use chromatography and mass spectrometry. The current PFAS detection method mainly focuses on blood, and there are few studies on urine, especially hair and nail PFAS detection methods. The existing methods have limitations such as complex pretreatment process, coexisting substance interference and unstable recovery rate, and a new detection method needs to be developed. SUMMARY

[0004] Therefore, the purpose of the present application is to provide a PFAS identification system and synchronous detection method based on human non-invasive biological samples (urine, hair, fingernail / toe nail) to solve the problems of insufficient precision and flux bottleneck in efficiently identifying and synchronously monitoring multiple key components of PFAS with high health hazards in the prior art.

[0005] The present application is achieved by the following technical solutions:

[0006] An identification system for human non-invasive biological matrix PFAS, the system establishment method comprising the following steps:

[0007] S1: Screening of key enterprise, industry, and regional characteristic pollutant PFAS;

[0008] S2: Screening of PFAS based on ecological risk, human health risk, and PMT / PBT substance characteristics of PFAS;

[0009] S3: Establishment of biological detection list based on the accumulation characteristics of PFAS in various non-invasive biological matrixes;

[0010] S4: Carrying out PFAS biological monitoring and prompting PFAS that need to be prioritized for control according to the human biological monitoring of PFAS.

[0011] Further limited, the S1 includes the following specific steps:

[0012] S101: Data collection, forming the key enterprise, industry, regional characteristic pollutants PFAS list I;

[0013] Through in-depth communication and docking with local environmental management departments and industry enterprises, the enterprise pollution discharge permit information is systematically sorted out, and the collected enterprise data is comprehensively analyzed and evaluated, and finally the key enterprises producing, using and discharging PFAS in the evaluation area are screened out, and the key enterprise characteristic pollutant PFAS list I is formed;

[0014] S102: Field investigation, forming the key enterprise characteristic pollutant PFAS list II;

[0015] After completing the preliminary screening, the PFAS contained in the raw and auxiliary materials in the production process of the enterprise, the potential intermediate products, conversion products and by-products in the process flow, the PFAS components in the products, and the PFAS discharged under abnormal conditions are focused on. After summarizing the above information, the key enterprise characteristic pollutant PFAS list II is formed;

[0016] S103: Detection and analysis, forming the key industry characteristic pollutant PFAS list III;

[0017] Representative environmental samples and food samples are taken, and PFAS detection and analysis are carried out on the collected samples to determine the key enterprise, industry, regional characteristic pollutant PFAS list III;

[0018] The S2 includes the following specific steps:

[0019] Constructing a comprehensive scoring method to screen characteristic pollutants PFAS;

[0020] Based on the following indicators, an index evaluation system of PFAS is constructed:

[0021] (1) The environmental exposure level (O) of PFAS is characterized by environmental detection concentration (O C ) and detection frequency (O DF );

[0022] (2) The ecological risk (E) of PFAS is characterized by ecological risk entropy (RQ); the calculation formula of RQ is:

[0023]

[0024] In the formula, MEC is the measured concentration of PFAS in the environmental medium, and PNEC is the predicted no-effect concentration of PFAS;

[0025] (3) The human health risk of PFAS is characterized by the lifetime cancer risk index (ILCR) and the hazard quotient (HQ), respectively, and the calculation formulas are as follows:

[0026] ILCR = ADD ing × SF ing + ADD der × SF der + ADD inh × SF inh

[0027]

[0028] wherein ADD ing , ADD der and ADD inh are the daily exposure doses through the digestive tract, skin contact and respiratory inhalation, respectively;

[0029] SF ing , SF der and SF inh are the carcinogenic slope factors through the digestive tract, skin contact and respiratory inhalation, respectively;

[0030] RfD ing , RfD der and RfD inh are the long-term intake reference doses through the digestive tract, skin contact and respiratory inhalation, respectively;

[0031] (4) The persistence (P) of PFAS is characterized by the biodegradation coefficient (BioWIN) based on the EPISuite model, the bioaccumulation (B) is characterized by the bioconcentration factor (BCF) based on the EPISuite model, and the migration ability (M) is characterized by the organic carbon-water partition coefficient (Koc) based on the EPISuite model;

[0032] The geometric grading method is adopted, and the grading standards are defined by the geometric progression. The index parameters O C , O DF , BioWIN, BCF, Koc, RQ, ILCR and HQ of the pollutants are divided into 5 geometric intervals according to the size and correspond to 5 grades respectively:

[0033] a n = a min × q n

[0034] wherein a n is the upper limit value of the nth interval of each evaluation parameter;

[0035] a min is the minimum value of each evaluation parameter;

[0036] q is a constant ratio;

[0037] n = 1, 2, 3, 4, 5;

[0038] The USEPA reporting quantity method is used to score each grade, that is, the data is transformed by the 2 / 3 cumulative rank method, the total score of each index is 100 points, and the score of grade 1 is the highest. See Table 1 for specific scoring. The environmental exposure score of the pollutant is the arithmetic mean of the environmental detection concentration score and the detection rate score. In the process of assigning scores to human health risks of pollutants, if the pollutant is a carcinogen, the carcinogenic risk score is taken, and if it is a non-carcinogen, the non-carcinogenic risk score is taken. The final screening obtained the list of industry characteristic pollutants PFAS IV.

[0039] Further limited, the PNEC is calculated based on the following method:

[0040] The acute toxicity data LC50 (half lethal concentration) or EC50 (maximum half-effect concentration) and chronic toxicity data NOEC (non-visible effect concentration) of each pollutant are obtained through the USEPA-ECOTOX toxicity database;

[0041] For new pollutants without PNEC data, the method for calculating PNEC in the European Union Chemical Risk Assessment Technical Guidance Document (TGD) is used to calculate the PNEC of water environment using the results of standard acute toxicity experiments;

[0042] The results of standard acute toxicity experiments include LC / EC50 and no-effect concentration (NOEC) of algae, water fleas and fish belonging to three different trophic level organisms;

[0043] Different trophic levels are selected to more comprehensively evaluate the impact of chemicals on each component of the ecosystem. When calculating PNEC, the toxicity test results of the most sensitive species are selected;

[0044] When using acute toxicity LC / EC50 data, the PNEC needs to be divided by an evaluation factor (AF) with a value of 1000;

[0045] When using chronic toxicity NOEC data, if NOEC data of 1, 2, or 3 trophic levels are available among the 3 trophic levels, the corresponding AF value is 100, 50, or 10.

[0046] Further limited, the daily exposure dose calculation formula through the digestive tract, skin contact and respiratory inhalation pathway is as follows:

[0047]

[0048] In the formula, ADD inhD is the daily average exposure amount of pollutants inhaled through the respiratory tract in ambient air, mg / (kg.d);

[0049] C a D is the concentration of pollutants inhaled through the respiratory tract in ambient air, mg / m 3 ;

[0050] IR is the respiratory volume, m 3 / h;

[0051] ET is the daily exposure time, h / d;

[0052] EF is the exposure frequency, d / a;

[0053] ED is the exposure duration, a;

[0054] BW is the body weight, kg;

[0055] AT is the average exposure time, d;

[0056] The daily average exposure amount taken in through the digestive tract includes the daily average exposure amount taken in through the mouth for food and the daily average exposure amount taken in through the mouth for drinking surface water or underground water and the daily average exposure amount taken in through the mouth for soil:

[0057] Wherein, the daily average exposure amount taken in through the mouth for food is calculated by the formula:

[0058]

[0059] In the formula, ADD oral-foad is the daily average exposure amount of pollutants taken in through the mouth for food, mg / (kg.d);

[0060] C f1 is the concentration of pollutants taken in through the mouth for food, mg / kg;

[0061] IR f1 is the food intake, kg / meal;

[0062] EF f1 is the food retention frequency, meals / a;

[0063] ED is the exposure duration, a;

[0064] BW is the body weight, kg;

[0065] AT is the average exposure time, d;

[0066] The daily average exposure amount taken in through the mouth for drinking surface water or underground water is calculated by the formula:

[0067]

[0068] In the formula, ADDoral-water AADD is the average daily dose of the contaminant from surface water or groundwater, mg / (kg·d);

[0069] C w CADD is the concentration of the contaminant in surface water or groundwater, mg / L;

[0070] IR w IRADD is the intake rate of surface water or groundwater, L / d;

[0071] EF is the exposure frequency, d / a;

[0072] ED is the exposure duration, a;

[0073] BW is the body weight, kg;

[0074] AT is the average time, d;

[0075] The formula for calculating the average daily dose of soil ingested orally is:

[0076]

[0077] where ADD oral-soil AADD is the average daily dose of the contaminant from soil ingested orally, mg / (kg·d);

[0078] C s CADD is the concentration of the contaminant in soil ingested orally, mg / kg;

[0079] IR s IRADD is the intake rate of soil, mg / d;

[0080] CF is the conversion factor, 1 x 10 6 kg / mg;

[0081] EF is the exposure frequency, d / a;

[0082] ED is the exposure duration, a;

[0083] BW is the body weight, kg;

[0084] AT is the average time, d;

[0085] The average daily dose of direct skin contact includes the average daily dose of soil ingested directly through the skin and the average daily dose of surface water or groundwater ingested through the skin;

[0086] where the formula for calculating the average daily dose of soil ingested directly through the skin is:

[0087]

[0088] where ADD dermal-soilDaily average exposure of the skin to the contaminant in soil, mg / (kg.d);

[0089] C s C is the concentration of the contaminant in soil, mg / kg;

[0090] CF is the mass conversion factor, 1 x 10 -6 kg / mg;

[0091] SA s SA is the surface area of the skin in contact with soil, cm 2 / event;

[0092] AF is the adhesion factor of the skin to soil, mg / cm 2 ;

[0093] ABS d ABS is the absorption factor of the skin to the contaminant, dimensionless;

[0094] EF is the frequency of the skin contact with soil, event / a;

[0095] ED is the duration of exposure, a;

[0096] BW is the body weight, kg;

[0097] AT is the average time of exposure, d;

[0098] The formula for the daily average exposure of the skin to surface water or groundwater is:

[0099]

[0100] where ADD dermal-water ADD is the daily average exposure of the skin to the contaminant in surface water or groundwater, mg / (kg.d);

[0101] C w C is the concentration of the contaminant in surface water or groundwater, mg / L;

[0102] SA w SA is the surface area of the skin in contact with surface water or groundwater, cm 2 ;

[0103] PC is the skin permeation constant of the contaminant, cm / h;

[0104] CF is the volume conversion factor, 1 x 10 -3 L / cm 3 ;

[0105] ET is the daily exposure time, h / d;

[0106] EF is the exposure frequency, d / a;

[0107] ED is the exposure duration, a;

[0108] BW is the body weight, kg;

[0109] AT is the average exposure time, d.

[0110] A synchronous detection method based on the above-mentioned human non-invasive biological matrix PFAS identification system, comprising the following steps:

[0111] Obtain the detected biological matrix, sequentially extract, purify and concentrate to obtain a detection sample, and analyze the detection sample by using an ultra-high performance liquid chromatograph mass spectrometer;

[0112] The liquid chromatography reference conditions are as follows:

[0113] The mobile phase is composed of A (5mM ammonium acetate) and B (acetonitrile);

[0114] The chromatographic conditions are as follows: flow rate 0.3mL / min, initial mobile phase 80% 5mM ammonium acetate aqueous solution (mobile phase A) and 20% acetonitrile (mobile phase B);

[0115] The elution gradient is as follows: 0min (%B), 9min (90%B), 12min (90%B), 12.10min (20%B) and 14min (20%B);

[0116] The total running time is 14min, the column temperature is 40℃, and the injection amount is 5μL.

[0117] The mass spectrometry reference conditions are as follows:

[0118] Electrospray ion source, negative ion mode;

[0119] Monitoring mode: multiple reaction monitoring;

[0120] Capillary voltage: 3000V; vacuum interface temperature: 200℃;

[0121] Desolvation gas temperature: 350℃;

[0122] Atomization gas flow: 1.0L / min;

[0123] Desolvation gas (nitrogen) flow: 15L / min;

[0124] Backflush gas flow: 1.5L / min;

[0125] Collision gas (argon) flow: 0.25ml / min.

[0126] Further limitation, the biological matrix is the nail, and the preparation method of the detection sample is:

[0127] Sample preparation, the nail sample was placed in a 50ml polypropylene (PP) centrifuge tube, and ultrapure water was ultrasonically cleaned for 10min; acetone was cleaned twice, and air was naturally dried;

[0128] The sample was crushed by a ball mill, 5000rpm, 3-5min;

[0129] Extraction, accurately weigh 0.1g of nail sample into a 50mL polypropylene (PP) centrifuge tube, add 2ng of extraction internal standard;

[0130] Add 10mL of 0.05mol / L NaOH-methanol digestion, room temperature oscillation for 8 hours; ultrasonic digestion at 55℃ for 30min, 9000xg centrifugation for 15min, the supernatant was filtered through a 0.22μm nylon membrane and transferred to a new 15mL PP centrifuge tube, nitrogen blowing (50℃); add 10mL of methanol to the residue, ultrasonic at 55℃ for 2h, centrifuge at 9000xg for 15min, and combine with the previous supernatant after filtration, continue to nitrogen blow to nearly dry (-2mL);

[0131] Concentrate the supernatant, add 500μL of 1M HCl to neutralize, and dilute with ultrapure water to about 50mL;

[0132] Purification, sequentially activate the SPE column (weak anion exchange solid phase extraction column, SelectCore WAX solid phase extraction column (200mg, 6mL) with 6mL of 2% ammonia-methanol, 6mL of methanol and 6mL of ultrapure water;

[0133] The extract was passed through the column at a flow rate of 3mL / min-5mL / min;

[0134] Vacuum drying for 10min to remove residual water in the column;

[0135] 6mL of 2% ammonia-methanol elution (flow rate of 1mL / min-3mL / min);

[0136] Concentrate, nitrogen blow to nearly dry, add 2ng of sample internal standard (PFOA-13C2), dilute to 1mL with methanol, mix well, filter through a 0.22μm needle filter into a sample bottle, seal, protect from light, and store at 4℃ or below.

[0137] Further limitation, the biological matrix is hair, and the preparation method of the detection sample is:

[0138] Sample preparation, the hair sample was placed in a 50ml polypropylene (PP) centrifuge tube, and ultrapure water was ultrasonically cleaned for 10min; acetone was cleaned twice, and air was naturally dried;

[0139] The sample was crushed by a ball mill, 5000rpm, 3-5min;

[0140] Extraction, accurately weigh 0.1 g of hair sample into a 50 mL polypropylene (PP) centrifuge tube, add 2 ng of extraction internal standard;

[0141] Add 10 mL of methanol-acetonitrile solution (v:v = 1:1), ultrasonic extraction at 55°C for 2 hours, centrifuge at 9000 x g for 25 minutes, transfer the supernatant to a 50 mL PP centrifuge tube, repeat the extraction for 2 times. The supernatant is filtered through a 0.22 μm nylon membrane into a new 15 mL PP centrifuge tube, and then is blown to near dryness (about 2 mL) under nitrogen (50°C). Add ultrapure water to dilute to about 50 mL;

[0142] Concentrate the supernatant, add 500 μL of 1 M HCl for neutralization, and then add ultrapure water to dilute to about 50 mL;

[0143] Purification, sequentially activate the SPE column (weak anion exchange solid phase extraction column, SelectCore WAX solid phase extraction column (200 mg, 6 mL) with 6 mL of 2% ammonia-methanol, 6 mL of methanol, and 6 mL of ultrapure water;

[0144] Pass the extract through the column at a flow rate of 3 mL / min to 5 mL / min;

[0145] Vacuum dry for 10 min to remove residual water in the column;

[0146] Elute with 6 mL of 2% ammonia-methanol at a flow rate of 1 mL / min to 3 mL / min;

[0147] Concentrate, blow to near dryness under nitrogen, add 2 ng of sample internal standard (PFOA-13C2), dilute to 1 mL with methanol, mix well, filter through a 0.22 μm needle filter into a sample bottle, seal, protect from light, and store at 4°C or below.

[0148] Further limitation, the biological matrix is urine, and the preparation method of the detection sample is as follows: the sampling amount of the urine sample is 1 mL, which is placed in a 50 mL polypropylene (PP) centrifuge tube, and 2 ng of extraction internal standard is added;

[0149] Extraction, add 1 mL of 1% formic acid, and ultrasonic extraction at 60°C for 30 min;

[0150] Purification, sequentially activate the SPE column (weak anion exchange solid phase extraction column, SelectCore WAX solid phase extraction column (200 mg, 6 mL) with 6 mL of 2% ammonia-methanol, 6 mL of methanol, and 6 mL of ultrapure water;

[0151] Pass the extract through the column at a flow rate of 3 mL / min to 5 mL / min;

[0152] Vacuum dry for 10 min to remove residual water in the column;

[0153] 6 mL 2% ammonia water methanol elution (flow rate of 1 mL / min ~ 3 mL / min);

[0154] Concentrate, nitrogen blow to near dry, add 2 ng injection internal standard (PFOA-13C2), with methanol constant volume to 1 mL, mix well, then filter through a 0.22 mu needle filter into a sample bottle, seal, avoid light, store at 4 DEG C or below.

[0155] Further limited, the extraction internal standard contains a mixed solution of 11 internal standards, specifically: PFBA- 13 C4, PFHxA- 13 C2, PFOA- 13 C4, PFNA- 13 C5, PFDA- 13 C2, PFUNDA- 13 C2, PFDoDA- 13 C2, PFHxS- 18 O2, PFOS- 13 C4, HFPODA- 13 C3, NEtFOSAA-d5.

[0156] The beneficial effects of the present application are:

[0157] The identification system and synchronous detection method of the human non-invasive biological matrix PFAS can realize directional identification and accurate monitoring of high-concern PFAS through urine, hair and nail PFAS detection, simple pretreatment process, convenient operation, low coexisting substance interference, high and efficient recovery rate, comprehensive evaluation system, coupling PFAS pollution source, environmental occurrence, human exposure and other factors, and scientific priority ranking method.

[0158] Other advantages, objects and features of the present application will be set forth in part in the following specification, and in part will become apparent to those skilled in the art upon examination of the following, or can be learned from practice of the present application. The objects and other advantages of the present application can be realized and attained by the methods and instrumentalities particularly pointed out in the following description. BRIEF DESCRIPTION OF DRAWINGS

[0159] Figure 1 The architecture diagram of the PFAS screening system for key enterprises, industries and regional characteristics; DETAILED DESCRIPTION

[0160] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. The components of the embodiments of the present application described and shown in the drawings can be arranged and designed in various different configurations.

[0161] Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the claimed application, but only represents selected embodiments of the present application. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without creative labor are within the scope of protection of the present application.

[0162] It should be noted that: similar reference numbers and letters represent similar items in the following drawings, so once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0163] In the above description of the present application, it should be noted that the terms "one side", "the other side" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, or the orientation or positional relationship in which the product of the present application is usually placed, which is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present application. In addition, the terms "first", "second" and the like are only used to distinguish the description and cannot be understood as indicating or implying relative importance.

[0164] In addition, the term "same" and other terms do not mean that the parts must be absolutely the same, but there can be slight differences. The term "vertical" only means that the positional relationship between the parts is more vertical than "parallel", and does not mean that the structure must be completely vertical, but can be slightly inclined.

[0165] Please refer to Figure 1 The present application provides a technical solution: a human non-invasive biological matrix PFAS identification system, the specific steps are as follows:

[0166] S1: Screening of key enterprises, industries, and regional characteristic pollutants PFAS;

[0167] S101: Data collection, forming a list of key enterprise characteristic pollutants PFAS I;

[0168] In the screening of regional key industries, first of all, industry investigation and research work is carried out. Through in-depth communication and docking with local environmental management departments and industry enterprises, the information of industry pollution discharge permit is systematically combed, and the collected industry data is comprehensively analyzed and evaluated, and finally the key enterprises producing and discharging PFAS in the regional scope are screened out, and the list of key enterprise characteristic pollutants PFAS I is formed.

[0169] S102: field investigation, forming the list of key enterprise characteristic pollutants PFAS II;

[0170] After completing the enterprise screening, the enterprise field investigation work is started immediately. In the process of enterprise field investigation, the contents such as organic matter contained in raw and auxiliary materials in the production process of enterprises, potential intermediate products and by-products in the process, organic components in products, and organic matter discharged under abnormal conditions are focused on. After summarizing the above information, the list of key enterprise characteristic pollutants PFAS II is formed. The research process mainly covers screening and determining research enterprises, comprehensively collecting enterprise status data, conducting on-site enterprise investigation, in-depth analysis of enterprise production and pollution discharge links, and on-site sampling, so as to realize the on-site investigation of the whole production process of enterprises, accurately locate the PFAS pollution discharge node, and complete the sampling work.

[0171] S103: detection and analysis, forming the list of key industry characteristic pollutants PFAS III;

[0172] Through professional detection and analysis of collected samples, the list of key industry characteristic pollutants PFAS III is determined.

[0173] S2: screening PFAS based on ecological risk, human health risk, and PMT / PBT substance characteristics of PFAS;

[0174] According to the classification of persistent, bioaccumulative and toxic substances (PBT) and persistent, mobile and toxic (PMT), the "SMILES" of the compound can be input on the online server (http: / / www.pmt.zj.cn / ) of PMT / PBT profiler to query the compound. The list of PFAS belonging to PMT / PBT is obtained by using this website.

[0175] Based on the following indicators, the index evaluation system of PFAS is constructed:

[0176] The environmental exposure level (O) of PFAS is characterized by environmental detection concentration (O C ) and detection frequency (O DF );

[0177] The persistence (P) is characterized by the biodegradation coefficient (BioWIN) based on the EPISuite model;

[0178] Bioaccumulation (B) is characterized by the bioconcentration factor (BCF) based on EPISuite model;

[0179] Migration ability (M) is characterized by the organic carbon-water partition coefficient (Koc) based on EPISuite model;

[0180] Ecological risk (E) of the pollutants is characterized by the ecological risk entropy (RQ); the calculation formula of RQ is

[0181]

[0182] In the formula, MEC is the measured concentration of PFAS in the environmental medium, and PNEC is the predicted no-effect concentration of PFAS. The PNEC is calculated based on the following method:

[0183] The acute toxicity data LC50 (half lethal concentration) or EC50 (maximum half-effect concentration) and the chronic toxicity data NOEC (non-observable effect concentration) of each pollutant are obtained through the USEPA-ECOTOX toxicity database. For new pollutants without PNEC data, the method for calculating PNEC in the European Union Technical Guidance Document (TGD) for chemical risk assessment is used to calculate the PNEC of the water environment by using the results of standard acute toxicology experiments. The standard acute toxicology experiment results include LC / EC50 and non-effect concentration (NOEC) of algae, water fleas and fish belonging to three different trophic levels. Different trophic levels are selected to more comprehensively evaluate the impact of chemicals on each component of the ecosystem. In the calculation of PNEC, the toxicology experiment results of the most sensitive species are generally selected (it is generally believed that the sensitivity of the ecosystem depends on the most sensitive species, and if the most sensitive species is protected, the structure of the ecosystem is also maintained in general balance). When using acute toxicity LC / EC50 data, the PNEC needs to be divided by an evaluation factor (AF) with a value of 1000; when using chronic toxicity NOEC data, if NOEC data of 1, 2, or 3 trophic levels are available among the 3 trophic levels, the corresponding AF value is 100, 50, or 10.

[0184] Health risk (H) can be divided into carcinogenic and non-carcinogenic risk assessment according to different modes of action of PFAS, and is characterized by the lifetime cancer risk index (ILCR) and the hazard entropy (HQ), respectively. The calculation formulas of ILCR and HQ are as follows:

[0185] ILCR = ADD ing × SF ing + ADD der × SF der + ADD inh × SF inh

[0186]

[0187] ADD ing , ADD der and ADD inh are the daily exposure doses via digestive tract, dermal contact and respiratory inhalation, respectively. SF ing , SF der and SF inh are the carcinogenic slope factors via digestive tract, dermal contact and respiratory inhalation, respectively. RfD ing , RfD der and RfD inh are the long-term intake reference doses via digestive tract, dermal contact and respiratory inhalation, respectively. Both SF and RfD are derived from the USEPA Integrated Risk Information System (IRIS) data.

[0188] The daily exposure dose calculation formulas via digestive tract, dermal contact and respiratory inhalation are as follows:

[0189]

[0190] ADD inh is the daily average exposure amount of pollutants inhaled through respiratory tract in ambient air, mg / (kg·d);

[0191] C a is the concentration of pollutants inhaled through respiratory tract in ambient air, mg / m 3 ;

[0192] IR is the respiratory volume, m 3 / h;

[0193] ET is the daily exposure time, h / d;

[0194] EF is the exposure frequency, d / a;

[0195] ED is the exposure duration, a;

[0196] BW is the body weight, kg;

[0197] AT is the average exposure time, d;

[0198] The daily average exposure amount via digestive tract includes the daily average exposure amount via oral intake of food, the daily average exposure amount via oral intake of surface water or underground water, and the daily average exposure amount via oral intake of soil:

[0199] The daily average exposure amount via oral intake of food is calculated by the formula:

[0200]

[0201] ADDoral-food Average daily exposure to a contaminant in food, mg / (kg.d);

[0202] C f1 Concentration of a contaminant in food, mg / kg;

[0203] IR f1 Food intake, kg / meal;

[0204] EF f1 Frequency of food intake, meals / a;

[0205] ED Exposure duration, a;

[0206] BW Body weight, kg;

[0207] AT Average time of exposure, d;

[0208] The formula for average daily exposure to a contaminant in surface or ground water consumed orally is:

[0209]

[0210] where ADD oral-water Average daily exposure to a contaminant in surface or ground water consumed orally, mg / (kg.d);

[0211] C w Concentration of a contaminant in surface or ground water, mg / L;

[0212] IR w Intake of surface or ground water, L / d;

[0213] EF Frequency of exposure, d / a;

[0214] ED Exposure duration, a;

[0215] BW Body weight, kg;

[0216] AT Average time of exposure, d;

[0217] The formula for average daily exposure to a contaminant in soil ingested orally is:

[0218]

[0219] where ADD oral-soil Average daily exposure to a contaminant in soil ingested orally, mg / (kg.d);

[0220] C s Concentration of a contaminant in soil, mg / kg;

[0221] IRs Dose for soil intake, mg / d;

[0222] CF is the conversion factor, 1 x 10 6 kg / mg;

[0223] EF is the exposure frequency, d / a;

[0224] ED is the exposure duration, a;

[0225] BW is the body weight, kg;

[0226] AT is the average time of exposure, d;

[0227] The daily average exposure amount of direct skin contact includes the daily average exposure amount of direct skin contact with soil and the daily average exposure amount of skin contact with surface water or groundwater;

[0228] The daily average exposure amount of direct skin contact with soil is calculated by the formula:

[0229]

[0230] In the formula, ADD dermal-soil The daily average exposure amount of skin contact with pollutants in soil, mg / (kg.d);

[0231] C s is the concentration of pollutants in soil contacted by skin, mg / kg;

[0232] CF is the conversion factor, 1 x 10 6 kg / mg;

[0233] SA s is the surface area of soil contacted by skin, cm 2 / event;

[0234] AF is the adhesion factor of skin to soil, mg / cm 2 ;

[0235] ABS d is the absorption factor of skin to pollutants, dimensionless;

[0236] EF is the frequency of skin contact with soil, event / a;

[0237] ED is the exposure duration, a;

[0238] BW is the body weight, kg;

[0239] AT is the average time of exposure, d;

[0240] The daily average exposure amount of skin contact with surface water or groundwater is calculated by the formula:

[0241]

[0242] where ADD dermal-water is the daily average exposure of the contaminant in surface water or groundwater to the skin, mg / (kg.d);

[0243] C w is the concentration of the contaminant in surface water or groundwater to the skin, mg / L;

[0244] SA w is the surface area of the skin exposed to surface water or groundwater, cm3;

[0245] PC is the skin permeation constant of the contaminant, cm / h;

[0246] CF is the volume conversion factor, 1 x 103L / cm3;

[0247] ET is the daily exposure time, h / d;

[0248] EF is the exposure frequency, d / a;

[0249] ED is the exposure duration, a;

[0250] BW is the body weight, kg;

[0251] AT is the average exposure time, d

[0252] The geometric grading method was used to define grading standards for each of the contaminant indicator parameters O C , O DF , BioWIN, BCF, Koc, RQ, ILCR and HQ using a geometric progression. Each of these parameters was divided into five equal intervals corresponding to five grades:

[0253] a n = a min x q n

[0254] where a n is the upper limit of the nth interval for each evaluation parameter; a min is the minimum value for each evaluation parameter; q is the geometric constant; and n = 1, 2, 3, 4, 5.

[0255] The USEPA's threshold of concern method was used to assign scores to each level, i.e., the data was transformed by the 2 / 3 cumulative rank method, and the total score of each indicator was 100 points, with the highest score for level 1. The environmental exposure score of the pollutant was the arithmetic mean of the environmental detection concentration score and the detection rate score. In the process of assigning scores to the human health risk of pollutants, if the pollutant was a carcinogen, the carcinogenic risk score was taken, and if it was a non-carcinogen, the non-carcinogenic risk score was taken. The final screening obtained the list of industry characteristic pollutants PFAS IV.

[0256] The following table is the grading and scoring of evaluation indicators

[0257] Table 1 PFAS evaluation grading and scoring table

[0258] In this study, equal weight was given to each evaluation indicator to calculate the comprehensive score of the pollutant:

[0259] S core-total = S core -O+S core -P+S core -B+S core -M+S core -E+S core -H

[0260] In the formula, Score-total, Score-O, Score-P, Score-B, Score-M, Score-E and Score-H

[0261] represent the comprehensive score of the pollutant, the environmental exposure level score, the persistence score, the bioaccumulation score, the migration ability score, the ecological risk score and the health risk score, respectively. The priority order of the pollutants was determined based on the high-low order of the comprehensive score of the pollutants, and the higher the comprehensive score, the greater the priority. Through the equal ratio grading method, the comprehensive scores of all pollutants were further divided into 6 levels (I ~ VI), and the comprehensive scores of pollutants in levels I ~ VI decreased in turn, and PFAS pollutants in levels I and II were listed as high priority pollutants.

[0262] S3: Establish a biological detection list based on the accumulation characteristics of PFAS in various non-creativity substrates

[0263] Urine: suitable for short-term monitoring of high-exposure populations, and can reflect the excretion of short-chain PFAS. The detection rate of perfluorocarboxylic acids below C7 and perfluorosulfonic acids below C6 is relatively high.

[0264] Hair: suitable for long-term monitoring of perfluoroalkyl substances with carbon chain length greater than 4, and may have a special accumulation tendency for PFOA.

[0265] Nail: More suitable for monitoring PFAS with carbon chain length greater than 4, due to the long growth cycle of nails, it can reflect the long-term load level in the body, especially suitable for long-term exposure assessment.

[0266] The simultaneous detection method of biologic matrix PFAS is as follows:

[0267] Sample pretreatment process

[0268] 1. Sample preparation

[0269] Hair or nail samples were placed in 50ml polypropylene (PP) centrifuge tubes, and ultrapure water (just cover the sample) was ultrasonically cleaned for 10min; acetone was cleaned 2 times, and the air was naturally dried;

[0270] The sample was crushed by a ball mill at 5000rpm for 3-5min;

[0271] 2. Extraction

[0272] 0.1g of hair / nail sample was accurately weighed and placed in a 50mL polypropylene (PP) centrifuge tube, and the sample amount of urine was 1ml; 2ng of extraction internal standard (extraction internal standard contains a mixed solution of 11 internal standards, which are PFBA- 13 C4, PFHxA- 13 C2, PFOA- 13 C4, PFNA- 13 C5, PFDA- 13 C2, PFUNDA- 13 C2, PFDoDA- 13 C2, PFHxS- 18 O2, PFOS- 13 C4, HFPODA- 13 C3, NEtFOSAA-d 5) .

[0273] a) Nail extraction

[0274] 10mL of 0.05mol / L NaOH-methanol was added for digestion, and the sample was oscillated at room temperature for 8 hours (overnight is recommended); ultrasonic digestion was carried out at 55°C for 30min, and centrifugation was carried out at 9000xg for 15min; the supernatant was filtered through a 0.22μm nylon membrane and transferred to a new 15mL PP centrifuge tube, and nitrogen blowing (50°C) was carried out; 10mL of methanol was added to the residue, and ultrasonic digestion was carried out at 55°C for 2h; centrifugation was carried out at 9000xg for 15min, and the filtrate was combined with the previous supernatant and continued to be nitrogen blown to near dryness (~2mL). The supernatant was concentrated, 500μL of 1M HCl was added for neutralization, and ultrapure water was added for dilution to about 50mL.

[0275] b) Hair extraction

[0276] Add 10 ml of methanol-acetonitrile solution (v:v = 1:1), ultrasonic extraction at 55°C for 2 hours, centrifugal at 9000 x g for 25 minutes, transfer the supernatant to a 50 mL PP centrifuge tube, repeat the extraction for 2 times. The supernatant is filtered through a 0.22 μm nylon membrane and transferred to a new 15 mL PP centrifuge tube, and then is blown to dryness (about 2 mL) under nitrogen (50°C). Add ultrapure water to dilute to about 50 mL.

[0277] c) Urine extraction

[0278] Add 1 ml of 1% formic acid, ultrasonic extraction at 60°C for 30 min.

[0279] 3. Purification

[0280] a) Activate the SPE column (weak anion exchange solid phase extraction column, SelectCore WAX solid phase extraction column (200 mg, 6 mL) with 6 mL of 2% ammonia water-methanol, 6 mL of methanol and 6 mL of ultrapure water in sequence;

[0281] b) The extract is passed through the column at a flow rate of 3 mL / min to 5 mL / min;

[0282] c) Vacuum drying for 10 min to remove residual water in the column;

[0283] d) Elute with 6 mL of 2% ammonia water-methanol (flow rate of 1 mL / min to 3 mL / min).

[0284] 4. Concentration

[0285] Blow to dryness under nitrogen, add sample internal standard, add 2 ng of sample internal standard (PFOA-13C2), dilute to 1 mL with methanol, mix well, filter through a 0.22 μm needle filter into a sample bottle, seal, protect from light, and store at 4°C or below.

[0286] 5. Sample detection process

[0287] Since the PFAS level in biological matrix such as hair, nails, etc. is trace or ultra-trace level, it is recommended to use ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS / MS) to analyze the sample. Establish a standard curve to calculate the average recovery and relative standard deviation of PFAS in the sample. When the recovery rate is within the range of 50-150%, it indicates that the method has good accuracy.

[0288] The standard curve was prepared by mixing a mixture solution containing 20 PFAS compound standards and 11 internal standard substances with a blank matrix to prepare a standard curve with concentrations of 0.30 ng / ml, 0.50 ng / ml, 1.00 ng / ml, 2.00 ng / ml, 4.00 ng / ml, 10.0 ng / ml, and 50.0 ng / ml. The 20 PFAS compounds are specifically perfluorobutyric acid (PFBA), perfluoropentanoic acid (PFPeA), perfluorohexanoic acid (PFHxA), perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUNDA), perfluorododecanoic acid (PFDoDA), perfluorotridecanoic acid (PFTrDA), perfluorotetradecanoic acid (PFTeDA), perfluorohexadecanoic acid (PFHxDA), perfluorobutane sulfonic acid (PFBS), perfluoropentane sulfonic acid (PFPeS), perfluorohexane sulfonic acid (PFHxS), perfluorooctane sulfonic acid (PFOS), perfluorononane sulfonic acid (PFNS), perfluorodecane sulfonic acid (PFDS), perfluoro-2-propoxypropanoic acid (HFPODA), and 11-chloro-3-oxa perfluoroundecane sulfonic acid (11Cl-PF3OUdS).

[0289] The quantitative analysis of the target substance was performed by ultra-high performance liquid chromatography (ACQUITY UPLC, Waters) combined with a triple quadrupole mass spectrometer (Xevo TQ-XS, Waters); the mass spectrometer was equipped with an electrospray ion source (ESI source), and the chromatographic column was selected as a Waters BEH C18 column (100 mm x 2.1 mm, 1.7 μm).

[0290] 1) Liquid chromatography reference conditions

[0291] The mobile phase was composed of A (5 mM ammonium acetate) and B (acetonitrile). The chromatographic conditions were as follows: flow rate 0.3 mL / min, initial mobile phase 80% 5 mM ammonium acetate aqueous solution (mobile phase A) and 20% acetonitrile (mobile phase B). The elution gradient was as follows: 0 min (%B), 9 min (90% B), 12 min (90% B), 12.10 min (20% B), and 14 min (20% B). The total running time was 14 min, the column temperature was 40°C, and the injection volume was 5 μL.

[0292] 2) Mass spectrometry reference conditions

[0293] Electrospray ion source, negative ion mode; monitoring mode: multiple reaction monitoring; capillary voltage: 3000 V; vacuum interface temperature: 200 °C; desolvation gas temperature: 350 °C; nebulizer gas flow: 1.0 L / min; desolvation gas (nitrogen) flow: 15 L / min; backflush gas flow: 1.5 L / min; collision gas (argon) flow: 0.25 ml / min.

[0294] Basic information of 20 PFASs is shown in Table 1, and MS / MS parameters are shown in Table 2.

[0295] Table 1 Basic information of perfluoroalkyl substances;

[0296]

[0297]

[0298] Table 2 MS / MS parameters for detection of 20 PFASs;

[0299]

[0300]

[0301] Through determination of the standard curve, the linear equation, correlation coefficient (R 2 ) of this method was determined, see Table 3.

[0302] Table 3 Standard curve and linear range of target substances;

[0303] Target Linear range (ng / ml) Standard curve [R 2 ]]> PFBA 0.3-50 y = 0.445619 * x - 0.026488 0.9946 PFPeA 0.3-50 y = 0.331246 * x + 0.0115764 0.9962 PFHxA 0.3-50 y = 0.608971 * x + 0.0128143 0.9994 PFHpA 0.3-50 y = 0.306353 * x + 0.00493496 0.9991 PFOA 0.3-50 y = 0.623583 * x + 0.0201119 0.9983 PFNA 0.3-50 y = 0.57436 * x + 0.0110994 0.9998 PFDA 0.3-50 y = 0.443981 * x - 0.0063039 0.9997 PFUnDA 0.3-50 y = 0.579469 * x - 0.0144773 0.9980 PFDoDA 0.3-50 y = 0.562779 * x + 0.00275842 0.9996 PFTrDA 0.3-50 y = 0.484671 * x + 0.0185816 0.9999 PFTeDA 0.3-50 y = 0.396536 * x + 0.0537831 0.9976 PFHxDA 0.3-50 y = 0.265316 * x + 0.0669585 0.9940 PFBS 0.3-50 y = 0.569129 * x + 0.00189276 0.9983 PFPeS 0.3-50 y = 0.486422 * x + 0.0190044 0.9983 PFHxS 0.3-50 y = 0.505363 * x + 0.0518079 0.9944 PFOS 0.3-50 y = 0.630661 * x + 0.021904 0.9944 PFNS 0.3-50 y = 0.56953 * x - 0.00236525 0.9995 PFDS 0.3-50 y = 0.475368 * x + 0.00454779 0.9997 HFPO-DA 0.3-50 y = 0.888075 * x - 0.0154102 0.9995 11Cl-PF3OUdS 0.3-50 y = 2.91229 * x + 0.135588 0.9960

[0304] Table 4 LOD and LOQ of 20 PFASs in three matrices;

[0305]

[0306]

[0307] The sample recovery rate of three types of non-traumatic biological matrixes, urine, nail and hair, is detected to evaluate the accuracy of the method. The specific operation is as follows: add low, medium and high concentration levels of target analyte, and 3 parallel samples for each concentration. Within one day, samples of three concentration levels are added for intraday evaluation (n=3); for two consecutive days, samples of three concentration levels of target substances are added for interday evaluation (n=6). The relative standard deviations (relative standard deviations, RSDs) of PFASs recovery rate are calculated to evaluate the intraday and interday precision, repeatability and reproducibility of the method. The test results are shown in Tables 5, 6 and 7. As can be seen from the tables, the average recovery rates of the three types of non-traumatic biological matrixes, urine, nail and hair, are within 50%-150% when the added concentrations are three low, medium and high concentration gradients, which has good accuracy and meets the analysis requirements.

[0308] Table 5 Average recovery rate, intraday and interday precision of different concentrations of target substances in urine

[0309]

[0310] As can be seen from the tables, when the added concentration of each substance is 0.5 ng / ml, the average recovery rate of the method is in the range of 68.3%-132.0%, the intraday RSD is 0.9%-23.5% (n=3), and the interday RSD is 3.2%-29.4% (n=6); when the added concentration of each substance is 5 ng / ml, the average recovery rate of the method is in the range of 64.8%-123.6%, the intraday RSD is 2.0-15.1% (n=3), and the interday RSD is 3.5%-14.8% (n=6); when the added concentration of each substance is 20 ng / ml, the average recovery rate of the method is in the range of 78.5%-118.9%, the intraday RSD is 0.8-15.0% (n=3), and the interday RSD is 1.2%-14.0% (n=6). The detection method of the present application has good accuracy, precision and reproducibility, and meets the analysis requirements of 20 kinds of PFASs in urine.

[0311] Table 6 Average recovery rate, intraday and interday precision of different concentrations of target substances in nail

[0312]

[0313] As shown in the table, when the spiked concentration of each substance is 10 ng / g, the average recovery of the method is in the range of 75.3%-144.2%, the intra-day RSD is 3.2%-29.8% (n=3), and the inter-day RSD is 4.0%-24.0% (n=6); when the spiked concentration of each substance is 20 ng / g, the average recovery of the method is in the range of 72.3%-115.8%, the intra-day RSD is 0.8%-13.0% (n=3), and the inter-day RSD is 2.2%-33.1% (n=6); and when the spiked concentration of each substance is 50 ng / g, the average recovery of the method is in the range of 69.9%-120.8%, the intra-day RSD is 2.0%-25.6% (n=3), and the inter-day RSD is 1.7%-17.8% (n=6). The detection method of the present application has good accuracy, precision and reproducibility, and meets the analysis requirements of 20 kinds of PFASs in nails.

[0314] Table 7 Average recovery, intra-day and inter-day precision of different concentrations of target substances in hair

[0315]

[0316]

[0317] As shown in the table, when the spiked concentration of each substance is 20 ng / g, the average recovery of the method is in the range of 92.8%-138.0%, the intra-day RSD is 1.9%-17.4% (n=3), and the inter-day RSD is 2.6%-12.1% (n=6); when the spiked concentration of each substance is 50 ng / g, the average recovery of the method is in the range of 75.4%-125.0%, the intra-day RSD is 1.2%-26.3% (n=3), and the inter-day RSD is 1.6%-21.1% (n=6); and when the spiked concentration of each substance is 100 ng / g, the average recovery of the method is in the range of 77.6%-126.9%, the intra-day RSD is 0.4%-24.2% (n=3), and the inter-day RSD is 1.0%-24.6% (n=6). The detection method of the present application has good accuracy, precision and reproducibility, and meets the analysis requirements of 20 kinds of PFASs in hair.

[0318] Finally, it should be pointed out that the above examples are only used to illustrate the technical solutions of the present application and not to limit the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced equivalently without departing from the purpose and scope of the technical solutions of the present application, and all of them should be covered in the scope of the claims of the present application.

Claims

1. A recognition system for PFAS (phosphoprotein oxidase inhibitors) in a non-invasive human biomatrix, characterized in that: The method for establishing this system includes the following steps: S1: Screening key enterprises, industries, and regions for characteristic pollutants PFAS; S2: Screening of PFAS based on ecological risk, human health risk, and PFAS properties of PMT / PBT substances; S3: Establish a biological detection list based on the accumulation characteristics of PFAS in various non-invasive biological matrices; S4: Conduct PFAS biomonitoring and identify PFAS that require priority control based on the human biomonitoring data.

2. The recognition system for non-invasive human biomatrix PFAS according to claim 1, characterized in that: S1 includes the following specific steps: S101: Data collection to form a list of characteristic pollutants PFAS of key enterprises, industries and regions (I); Through in-depth communication and coordination with local environmental management departments and industry enterprises, we systematically sorted out the enterprises' pollutant discharge permit information, and conducted a comprehensive analysis and evaluation of the collected enterprise data. Finally, we selected key enterprises that produce, use, and discharge PFAS within the assessment area and formed a list of characteristic pollutants PFAS of key enterprises (List I). S102: Conduct on-site investigations and compile a list of key enterprises' characteristic pollutants PFAS II; After completing the initial screening, focus on the PFAS contained in the raw and auxiliary materials of the enterprise's production process, the potential intermediate products, conversion products and by-products in the process flow, the PFAS components present in the products, and the PFAS emitted under abnormal operating conditions. After summarizing and organizing the above information, a list of characteristic pollutants PFAS of key enterprises is formed II. S103: Detection and analysis to form a PFAS inventory of key industry characteristic pollutants III; Representative environmental and food samples were collected, and PFAS analysis was conducted on the collected samples to determine the PFAS list III of key enterprises, industries, and regions. S2 includes the following specific steps: Construct a comprehensive scoring method to screen characteristic pollutant PFAS; The following indicators are used to construct an evaluation system for PFAS: (1) Environmental exposure level (O2) of PFAS, expressed as environmental detection concentration (O2). C ) and detection frequency (O DF Characterization; (2) The ecological risk (E) of PFAS is characterized by the ecological risk entropy (RQ); the formula for calculating RQ is: In the formula, MEC is the measured concentration of PFAS in the environmental medium, and PNEC is the predicted no-effect concentration of PFAS. (3) The human health risks of PFAS are characterized by the lifetime carcinogenic risk index (ILCR) and hazard entropy (HI), respectively, and the calculation formulas are as follows: ILCR=ADD ing ×SF ing +ADD der ×SF der +ADD inh ×SF inh In the formula, ADD ing ADD der and ADD inh These are the daily exposure doses via the digestive tract, skin contact, and inhalation routes, respectively. SF ing SF der and SF inh These are carcinogenic slope factors that occur via the digestive tract, skin contact, and inhalation routes, respectively. RfD ing 、RfD der and RfD inh These are the long-term intake reference doses for exposure via the digestive tract, skin contact, and inhalation routes, respectively. (4) The persistence (P) of PFAS was characterized by the biodegradation coefficient (BioWIN) obtained based on the EPISUite model; the bioaccumulation (B) was characterized by the bioaccumulation factor (BCF) obtained based on the EPISUite model; and the migration capacity (M) was characterized by the organic carbon-water partition coefficient (Koc) obtained based on the EPISUite model. The geometric classification method is adopted, and the classification criteria are defined using a geometric series to classify the various index parameters of pollutants. C O DF BioWIN, BCF, Koc, RQ, ILCR, and HQ are each divided into 5 equal intervals based on size, and each corresponds to 5 levels: a n =a min ×q n In the formula, a n This represents the upper limit of the nth level interval for each evaluation parameter; a min The minimum value of each evaluation parameter; q is a geometric constant; n＝1，2，3，4，5； The USEPA's reporting requirement method was used to assign scores to each level, specifically by converting the data using the 2 / 3 cumulative rank method. The total score for each indicator was 100 points, with level 1 having the highest score. The specific scoring is shown in Table 1. The environmental exposure score for pollutants was the arithmetic mean of the environmental detection concentration score and the detection rate score. In the process of assigning human health risk scores for pollutants, if the pollutant was a carcinogen, the carcinogenic risk score was used; if it was a non-carcinogen, the non-carcinogenic risk score was used. Finally, a list of industry-specific PFAS pollutants IV was obtained.

3. The recognition system for non-invasive human bio-matrix PFAS according to claim 2, characterized in that: The PNEC is calculated based on the following method: Acute toxicity data (LC50, or half-lethal concentration) and chronic toxicity data (NOEC, or no-effect concentration) for each pollutant were obtained from the USEPA-ECOTOX toxicity database. For new pollutants for which no PNEC data is available, the PNEC of the aquatic environment is calculated using the method for calculating PNEC in the EU Technical Guidance Document on Risk Assessment of Chemical Substances (TGD) and the results of standard acute toxicology tests. Standard acute toxicology test results include LC / EC50 and no-effect concentration (NOEC) for organisms belonging to three different trophic levels: algae, daphnia, and fish. Different trophic levels were selected to more comprehensively assess the impact of chemicals on all components of the ecosystem. When calculating PNEC, the toxicological results of the most sensitive species were used. When using acute toxicity LC / EC50 data, the PNEC needs to be divided by the evaluation coefficient (AF) with a value of 1000. When using chronic toxicity NOEC data, if NOEC data for 1, 2, and 3 of the 3 trophic levels are available, the corresponding AF values ​​are 100, 50, and 10, respectively.

4. The recognition system for non-invasive human bio-matrix PFAS according to claim 3, characterized in that: The formulas for calculating daily exposure doses via the digestive tract, skin contact, and inhalation routes are as follows: In the formula, ADD inh The daily average exposure to pollutants in ambient air inhaled through the respiratory tract, in mg / (kg·d); C a The concentration of pollutants in ambient air inhaled through the respiratory tract, in mg / m³ 3 ; IR stands for respiratory volume, m 3 / h; ET represents daily exposure time, in hours (h / d). EF represents the exposure frequency, in days per year (d / a). ED represents the duration of exposure, a; BW stands for body weight, in kg; AT represents the average exposure time, expressed in days (d). Daily exposure via the digestive tract includes daily exposure via oral food, daily exposure via oral drinking of surface or groundwater, and daily exposure via oral soil: The formula for calculating the average daily exposure to food ingested orally is as follows: In the formula, ADD oral-food The daily average exposure to contaminants in food via oral ingestion is expressed in mg / (kg·d). C f1 The concentration of contaminants in food ingested orally, in mg / kg; IR f1 Food intake, kg / meal; EF f1 Meals / a is the frequency of food retention. ED represents the duration of exposure, a; BW stands for body weight, in kg; AT represents the average exposure time, expressed in days (d). The formula for calculating the average daily exposure to surface water or groundwater via oral consumption is as follows: In the formula, ADD oral-water The daily average exposure to pollutants in surface water or groundwater via oral consumption is expressed in mg / (kg·d). C w The concentration of pollutants in surface water or groundwater intended for oral consumption is expressed in mg / L. IR w Surface water or groundwater intake, in L / d; EF represents the exposure frequency, in days per year (d / a). ED represents the duration of exposure, a; BW stands for body weight, in kg; AT represents the average exposure time, expressed in days (d). The formula for calculating the average daily exposure to soil through oral ingestion is as follows: In the formula, ADD oral-soil The daily average exposure to pollutants in the soil via oral ingestion is expressed in mg / (kg·d). C s The concentration of pollutants in the soil ingested orally is expressed in mg / kg. IR s Soil intake, mg / d; CF is the quality conversion factor, 1×10⁻⁶. 6 kg / mg; EF represents the exposure frequency, in days per year (d / a). ED represents the duration of exposure, a; BW stands for body weight, in kg; AT represents the average exposure time, expressed in days (d). Daily exposure to direct skin contact includes daily exposure to direct skin contact with soil and daily exposure to direct skin contact with surface water or groundwater. The formula for calculating the average daily exposure to soil through direct skin contact is as follows: In the formula, ADD dermal-soil Average daily exposure to pollutants in soil through skin contact, mg / (kg·d); C s This refers to the concentration of pollutants in soil upon skin contact, in mg / kg. CF is the quality conversion factor, 1×10 -6 kg / mg; SA s The surface area of ​​skin in contact with soil, in cm² 2 / event; AF is the skin-to-soil adhesion factor, mg / cm³. 2 ; ABS d It is a dimensionless factor that is the skin's absorption factor for pollutants. EF represents the frequency of skin contact with soil, in events per year. ED represents the duration of exposure, a; BW stands for body weight, in kg; AT represents the average exposure time, expressed in days (d). The formula for calculating the average daily exposure to surface water or groundwater through skin contact is as follows: In the formula, ADD dermal-water The average daily exposure to pollutants in surface water or groundwater via skin contact, in mg / (kg·d); C w The concentration of pollutants in surface water or groundwater that comes into contact with the skin is measured in mg / L. SA w The surface area of ​​skin in contact with surface water or groundwater, in cm² 2 ; PC represents the skin permeability constant of contaminants, in cm / h; CF is the volume conversion factor, 1×10 -3 L / cm 3 ; ET represents daily exposure time, in hours (h / d). EF represents the exposure frequency, in days per year (d / a). ED represents the duration of exposure, a; BW stands for body weight, in kg; AT represents the average exposure time, in days.

5. A synchronous detection method based on the recognition system of PFAS (non-invasive human biomatrix) according to any one of claims 1 to 4, characterized in that: The steps include the following: The biological matrix to be tested was obtained, and the sample was extracted, purified and concentrated in sequence to obtain the test sample. The test sample was then analyzed using ultra-high performance liquid chromatography-mass spectrometry. The reference conditions for liquid chromatography are as follows: The mobile phase consists of A (5mM ammonium acetate) and B (acetonitrile); The chromatographic conditions were as follows: flow rate 0.3 mL / min, initial mobile phase was 80% 5 mM ammonium acetate aqueous solution (mobile phase A) and 20% acetonitrile (mobile phase B); The elution gradients were: 0 min (%B), 9 min (90%B), 12 min (90%B), 12 min (20%B), 10 min (20%B), and 14 min (20%B). The total run time was 14 min, the column temperature was 40℃, and the injection volume was 5 μL. The reference conditions for mass spectrometry are: Electrospray ion source, negative ion mode; Monitoring method: Multiple response monitoring; Capillary voltage: 3000V; Vacuum interface temperature: 200℃; Desolvent gas removal temperature: 350℃; Atomizing gas flow rate: 1.0 L / min; Desolvent gas (nitrogen) flow rate: 15 L / min; Backflush air flow rate: 1.5 L / min; Collision gas (argon) flow rate: 0.25 ml / min.

6. The synchronous detection method according to claim 5, characterized in that: The biological matrix is ​​fingernail, and the sample preparation method is as follows: For sample preparation, the nail sample was placed in a 50ml polypropylene (PP) centrifuge tube and ultrasonically cleaned with ultrapure water for 10 minutes; then cleaned twice with acetone and allowed to air dry naturally. The sample was pulverized using a ball mill at 5000 rpm for 3–5 minutes. Extraction: Accurately weigh 0.1g of nail sample into a 50mL polypropylene (PP) centrifuge tube, and add 2ng of extraction internal standard; Add 10 mL of 0.05 mol / L NaOH-methanol for digestion, and shake at room temperature for 8 hours; sonicate at 55 °C for 30 min, centrifuge at 9000 × g for 15 min, filter the supernatant through a 0.22 μm nylon membrane and transfer it to a new 15 mL PP centrifuge tube, and blow with nitrogen (50 °C); add 10 mL of methanol to the residue, sonicate at 55 °C for 2 h, centrifuge at 9000 × g for 15 min, filter and combine with the previous supernatant, and continue to blow with nitrogen until nearly dry (~2 mL); Concentrate the supernatant, add 500 μL of 1M HCl to neutralize, and dilute with ultrapure water to about 50 mL. Purification was performed by activating the SPE column (weak anion exchange solid phase extraction column, SelectCoreWAX solid phase extraction column (200mg, 6mL)) sequentially with 6mL of 2% ammonia-methanol, 6mL of methanol, and 6mL of ultrapure water. The extract was passed through the column at a flow rate of 3 mL / min to 5 mL / min; Vacuum dry for 10 minutes to remove residual moisture from the column; Elute with 6 mL of 2% ammonia water and methanol (flow rate of 1 mL / min to 3 mL / min); Concentrate the solution, blow it with nitrogen until nearly dry, add 2 ng of internal standard (PFOA-13C2), dilute to 1 mL with methanol, mix well, filter through a 0.22 μm syringe filter into a sample vial, seal, protect from light, and store at 4°C or below.

7. The synchronous detection method according to claim 5, characterized in that: The biological matrix is ​​hair, and the sample preparation method is as follows: Sample preparation: hair samples were placed in 50ml polypropylene (PP) centrifuge tubes, ultrasonically cleaned with ultrapure water for 10 min; washed twice with acetone, and air-dried naturally. The sample was pulverized using a ball mill at 5000 rpm for 3–5 minutes. Extraction: Accurately weigh 0.1g of hair sample into a 50mL polypropylene (PP) centrifuge tube, and add 2ng of extraction internal standard; Add 10 mL of methanol-acetonitrile solution (v:v = 1:1), sonicate at 55 °C for 2 hours, centrifuge at 9000 × g for 25 minutes, transfer the supernatant to a 50 mL PP centrifuge tube, and repeat the extraction twice; filter the supernatant through a 0.22 μm nylon membrane and transfer it to a new 15 mL PP centrifuge tube, blow it with nitrogen (50 °C) until nearly dry (~2 mL), and dilute with ultrapure water to about 50 mL; Concentrate the supernatant, add 500 μL of 1M HCl to neutralize, and dilute with ultrapure water to about 50 mL. Purification was performed by activating the SPE column (weak anion exchange solid phase extraction column, SelectCoreWAX solid phase extraction column (200mg, 6mL)) sequentially with 6mL of 2% ammonia-methanol, 6mL of methanol, and 6mL of ultrapure water. The extract was passed through the column at a flow rate of 3 mL / min to 5 mL / min; Vacuum dry for 10 minutes to remove residual moisture from the column; Elute with 6 mL of 2% ammonia water and methanol (flow rate of 1 mL / min to 3 mL / min); Concentrate the solution, blow it with nitrogen until nearly dry, add 2 ng of internal standard (PFOA-13C2), dilute to 1 mL with methanol, mix well, filter through a 0.22 μm syringe filter into a sample vial, seal, protect from light, and store at 4°C or below.

8. The synchronous detection method according to claim 5, characterized in that: The biological matrix is ​​urine. The sample preparation method is as follows: 1 ml of urine sample is placed in a 50 mL polypropylene (PP) centrifuge tube, and 2 ng of internal standard is added for extraction. Extract by adding 1 ml of 1% formic acid and ultrasonic extraction at 60°C for 30 min. Purification was performed by activating the SPE column (weak anion exchange solid phase extraction column, SelectCoreWAX solid phase extraction column (200mg, 6mL)) sequentially with 6mL of 2% ammonia-methanol, 6mL of methanol, and 6mL of ultrapure water. The extract was passed through the column at a flow rate of 3 mL / min to 5 mL / min; Vacuum dry for 10 minutes to remove residual moisture from the column; Elute with 6 mL of 2% ammonia water and methanol (flow rate of 1 mL / min to 3 mL / min); Concentrate the solution, blow it with nitrogen until nearly dry, add 2 ng of internal standard (PFOA-13C2), dilute to 1 mL with methanol, mix well, filter through a 0.22 μm syringe filter into a sample vial, seal, protect from light, and store at 4°C or below.

9. The synchronous detection method according to any one of claims 6 to 8, characterized in that: The extracted internal standard comprises a mixed solution of 11 internal standards, specifically: PFBA- 13 C4, PFHxA- 13 C2, PFOA- 13 C4, PFNA- 13 C5, PFDA 13 C2, PFUNDA- 13 C2, PFDoDA- 13 C2、PFHxS- 18 O2, PFOS 13 C4, HFPODA- 13 C3, NETFOSAA-d5.

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