# US 2024/0201176 A1 · Method and device for synchronous extraction and analysis of multiple trace emerging organic contaminants in hairs, and application

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- Number: US 2024/0201176 A1
- Document ID: US20240201176A1
- Status: Published application
- Kind: A1
- Application number: 18/334,365
- Filed: 2023-06-13
- Published: 2024-06-20
- Earliest claimed: 2022-12-15
- Assignees: South China Inst of Environmental Sciences Mee Ecology and Environment Emergency Inst of Ecology
- Applicants: South China Inst of Environmental Sciences Mee Ecology and Environment Emergency Inst of Ecology
- Inventors: Jing ZHENG; Rongfa XU; Bin TANG; Xi ZHUANG; Xiao YAN; Weikeng LUO; Yunjiang YU
- Web page: https://patentgrep.com/patent/US20240201176A1

## CPC

- G01N 1/28: Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00})
- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G01N 1/286: {involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F)}
- G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)}
- G01N 2001/2866: {Grinding or homogeneising}
- G01N 2030/062: {extracting sample from raw material}
- G01N 2430/00: Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
- G01N 2560/00: Chemical aspects of mass spectrometric analysis of biological material
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A method for synchronous extraction and analysis of multiple trace emerging organic contaminants in hairs, and application comprises the following steps: S1, pretreatment; S2, mixing and extraction; S3, nitrogen blowing concentration and purification; and S4, low-temperature separation. A device for synchronous extraction and analysis of multiple trace emerging organic contaminants in hairs, and application comprises comprises a treatment barrel, a pedestal and a drive assembly. This method is applied to a study on long-term exposure of multiple organic contaminants in a human body. According to the present disclosure, an extraction solution extracting system is established, 5 emerging organic contaminants in hairs can be purified and analyzed by establishing a dispersion solid-phase extraction filler consisting of Na2SO4 powders and C18 powders.

## Claims

### Claim 1

A method for synchronous extraction and analysis of multiple trace emerging organic contaminants in hairs, comprising the following steps:
S1, pretreatment: adding a hair sample to be treated together with ultrapure water into a treatment barrel (1) in a weight ratio of 1:20-40, cutting the hair sample through a cutting quartz knife (62) in the treatment barrel (1) while washing for 0.5-1 h, subsequently discharging ultrapure water and adding ultrapure whose weight is 20-40 times of the weight of the hair sample again, cutting the hair sample through the cutting quartz knife (62) in the treatment barrel (1) while washing, until the length of the hair sample is 0.5-1 cm, subsequently discharging the ultrapure water, and then grinding the hair sample for 5-10 min through a grinding plate (11) in the treatment barrel (1) to obtain hair powders;
S2, mixing and extraction: drying the hair powders obtained in step S1, placing 0.1 g of dried hair powders into a centrifuge tube, adding 1 mL of ethyl acetate and 1 mL of internal standard indicator into the centrifuge tube, standing for 12 h, and then adding 4 mL of extract solution into the centrifuge tube, wherein the extract solution is prepared from n-hexane, acetone, acetonitrile and ethyl acetate in a volume ratio of 1:1:1:1, and then starting to perform centrifugal extraction, wherein the steps for centrifugal extraction are as follows: vortex treatment is carried out for 4 min at the rotation speed of 2000 r·min−1, then ultrasonic treatment is carried out for 20 min at the temperature of 20° C., and finally centrifugal treatment is carried out for 15 min at the rotation speed of 4000 r·min−1, centrifugal extraction is repeated 3 times, and then the obtained supernatant is merged as a supernatant raw solution;
S3, nitrogen blowing concentration and purification: performing nitrogen blowing concentration on the supernatant raw solution obtained in step S2 to 1 mL, subsequently adding 20 mg of Na2SO4 powders and 100 mg of C18 powders serving as purifying fillers, then performing vortex treatment for 7 min at the rotation speed of 1200 r·min−1, then performing centrifugal treatment for 15 min at the rotation speed of 3500 r·min−1 to obtain a concentrated solution, and then transferring the concentrated solution to a glass tube; and
S4, low-temperature separation: blowing the concentrated solution obtained in step S3 with nitrogen for 1-2 h, then adding 200 μL of methanol solution with a mass fraction of ≥99.8% for redissolving, standing for 4-6 h at the temperature of −20° C., freezing and removing a precipitate to obtain the hair raw solution after low-temperature separation and performing Liquid chromatography-tandem mass spectrometry/mass spectrometry (LC-MS/MS) and Gas chromatography-tandem mass spectrometry/mass spectrometry (GC-MS/MS) detection analysis on the hair raw solution.

### Claim 2 (depends on 1)

The method according to claim 1, wherein a method for preparing the internal standard indicator in the step S2 is as follows: mixing 0.1 mL of 50 μg/mL first internal standard raw solution, 0.1 mL of 50 μg/mL second internal standard raw solution, 0.05 mL of 100 μg/mL third internal standard raw solution, 0.05 mL of 100 μg/mL fourth internal standard raw solution and 0.05 mL of 100 μg/mL fifth internal standard raw solution and then adding 0.65 mL of methanol solution to obtain 1 mL of 5 μg/mL mixed internal standard, wherein the first internal standard raw solution is formed by mixing MPFOS and MPFOA in a volume ratio of 1:1, the second internal raw solution is formed by mixing d18-TCIPP, d15-TDCIPP, d15-TPhP and d12-TCEP in a volume ratio of 1:1:1:1, the third internal standard raw solution is formed by mixing d4-DEHP with d4-DNBP in a volume ratio of 1:1, the fourth internal standard raw solution is formed by mixing 13C4-mmBP, 13C4—OH-mEHP, 13C-mEP, 13C-mBzP, d4-mmOP, 50xo-MEHP-13C and MEHP-13C4 in a volume ratio of 1:1:1:1:1:1:1, and the fifth internal standard raw solution is formed by mixing BDE118, BDE128 and 13C12-BDE209 in a volume ratio of 1:1:1; and then diluting the mixed internal standard with a methanol solution to obtain an 100 ng/mL internal standard indicator.

### Claim 3 (depends on 2)

A device for use in the method according to claim 2, comprising a treatment barrel (1) for washing, cutting and grinding a hair sample, a pedestal (2) for supporting the treatment barrel (1), and a drive assembly (3) located above the treatment barrel (1);
wherein, the middle inside the treatment barrel (1) is provided with a rotating outer cylinder (4), the inside of the rotating outer cylinder (4) is slidaly sleeved with a rotating inner cylinder (5) that synchronously rotates with the rotating outer cylinder (4), the external circumference of the rotating outer cylinder (4) is provided with a plurality of groups of hair cutting assemblies (6) at equal intervals, the hair cutting assembly (6) includes two auxiliary quartz knives (61) and two cutting quartz knives (62), the auxiliary quartz knives (61) and the cutting quartz knives (62) are alternately arranged, the auxiliary quartz knives (61) are arranged at the bottom and middle of the outer wall of the rotating outer cylinder (4), the upper and lower sides of the auxiliary quartz knives (61) arranged in the middle of the outer wall of the rotating outer cylinder (4) are both provided with sliding grooves (41), a connection block (51) is arranged at the position of the sliding groove (41) corresponding to the outer wall of the rotating inner cylinder (5), the connection block (51) up and down slides along the sliding groove (41), the outer end of the connection block (51) is fixedly connected with the cutting quartz knives (62), the auxiliary quartz knives (61) and the cutting quartz knives (62) both extend to the inner wall close to the treatment barrel (1), the auxiliary quartz knives (61) and the cutting quartz knives (62) on the same group of cutting assemblies (6) are seamlessly engaged and aligned with each other through the blades (63), the blades (63) provided on the cutting quartz knives (62) are alternately engaged and aligned with the blades (63) provided correspondingly on the auxiliary quartz knives (61) when sliding up and down, the bottom of the treatment barrel (1) is provided with a grinding plate (11), and the two sides of the bottoms of the auxiliary quartz knives (61) located at the bottom of the outer wall of the rotating outer cylinder (4) are provided with grinding portions (42) that are matched with the grinding plate (11) to grind the hair samples;
the top of the treatment barrel (1) is provided with a top cover (12), the top of the rotating inner cylinder (5) penetrates through the top cover (12) and then is connected with the output end of a rotating motor (31) of the drive assembly (3), the output end of the rotating motor (31) is provided with a sleeve (32) that is in up and down sliding connection with the rotating inner cylinder (5), the outer wall of the rotating inner cylinder (5) located above the top cover (12) is provided with a cylindrical telescopic gear lever (52), the telescopic gear lever (52) includes an inner rod (53) and an outer rod (54) that are slidably sleeved, the upper surface of the top cover (12) close to the edge is circumferentially provided with a plurality of bumps (13) at equal intervals, the top surface of the bump (13) is arc-shaped, the inner rod (53) corresponds to the bump (13) after extending to lift the telescopic gear lever (52), one side of the upper surface of the top cover (12) is provided with an opening (14) for delivering hair samples and ultrapure water, and the bottom of the outer side wall of the treatment barrel (1) is provided with a drainage outlet (15).

### Claim 4 (depends on 3)

The device according to claim 3, wherein a total of 6 groups of hair cutting assemblies (6) are provided, the tail ends of the auxiliary cutting quartz knife (61) and the cutting quartz knife (62) are aligned, the surface of the cutting quartz knife (62) is provided with a plurality of permeable pores (64), the surface of the auxiliary cutting quartz knife (61) is provided with a slot (65), a total of 6 bumps are provided, the auxiliary cutting quartz knife (61) is connected with a clamp groove (44) arranged on the outer wall of the rotating outer cylinder (4) through a fixation block (43), the top and bottom of the fixation block (43) are respectively connected with the top surface and bottom surface of the clamp groove (44) through a first spring rod (45), the two sides of the outer wall of the rotating inner cylinder (5) are provided with limit strips (55), the limit strips (55) are both slidably connected with first limit grooves (46) arranged at the two sides of the inside of the rotating outer cylinder (4) and second limit grooves (33) arranged at the two sides of the inner wall of the sleeve (32), the bottom of the rotating inner cylinder (5) is provided with a weighting block (56), the bottom of the weighting block (56) is connected with the inner bottom of the rotating outer cylinder (4) through a second spring rod (57).

### Claim 5 (depends on 3)

The device according to claim 3, wherein the two sides of the upper surface of the pedestal (2) are symmetrically provided with struts (21), the top of the strut (21) is fixedly connected with the rotating motor (31) through one connection rod (22), the junction between the connection rod (22) and the rotating motor (31) is provided with a damping spacer (34), the telescopic rods (35) are arranged below the two connection rods (22), the bottoms of the two telescopic rods (35) are provided with a limit plate (36) of the inner rod (53) for downward pressing the telescopic gear lever (52), the tail end of the outer rod (54) is provided with a limit hole (58), the tail end of the inner rod (53) is provided with a limit protrusion (59) abutted against the limit hole (58), 3 insulation warehouses (23) are arranged inside the pedestal (2), temperature adjusters (24) are arranged inside the insulation warehouses (23), and the two sides of the upper surface of the pedestal (2) are each provided with an observation beaker (25).

### Claim 6 (depends on 1)

A device for use in the method according to claim 1, comprising a treatment barrel (1) for washing, cutting and grinding a hair sample, a pedestal (2) for supporting the treatment barrel (1), and a drive assembly (3) located above the treatment barrel (1);
wherein, the middle inside the treatment barrel (1) is provided with a rotating outer cylinder (4), the inside of the rotating outer cylinder (4) is slidaly sleeved with a rotating inner cylinder (5) that synchronously rotates with the rotating outer cylinder (4), the external circumference of the rotating outer cylinder (4) is provided with a plurality of groups of hair cutting assemblies (6) at equal intervals, the hair cutting assembly (6) includes two auxiliary quartz knives (61) and two cutting quartz knives (62), the auxiliary quartz knives (61) and the cutting quartz knives (62) are alternately arranged, the auxiliary quartz knives (61) are arranged at the bottom and middle of the outer wall of the rotating outer cylinder (4), the upper and lower sides of the auxiliary quartz knives (61) arranged in the middle of the outer wall of the rotating outer cylinder (4) are both provided with sliding grooves (41), a connection block (51) is arranged at the position of the sliding groove (41) corresponding to the outer wall of the rotating inner cylinder (5), the connection block (51) up and down slides along the sliding groove (41), the outer end of the connection block (51) is fixedly connected with the cutting quartz knives (62), the auxiliary quartz knives (61) and the cutting quartz knives (62) both extend to the inner wall close to the treatment barrel (1), the auxiliary quartz knives (61) and the cutting quartz knives (62) on the same group of cutting assemblies (6) are seamlessly engaged and aligned with each other through the blades (63), the blades (63) provided on the cutting quartz knives (62) are alternately engaged and aligned with the blades (63) provided correspondingly on the auxiliary quartz knives (61) when sliding up and down, the bottom of the treatment barrel (1) is provided with a grinding plate (11), and the two sides of the bottoms of the auxiliary quartz knives (61) located at the bottom of the outer wall of the rotating outer cylinder (4) are provided with grinding portions (42) that are matched with the grinding plate (11) to grind the hair samples;
the top of the treatment barrel (1) is provided with a top cover (12), the top of the rotating inner cylinder (5) penetrates through the top cover (12) and then is connected with the output end of a rotating motor (31) of the drive assembly (3), the output end of the rotating motor (31) is provided with a sleeve (32) that is in up and down sliding connection with the rotating inner cylinder (5), the outer wall of the rotating inner cylinder (5) located above the top cover (12) is provided with a cylindrical telescopic gear lever (52), the telescopic gear lever (52) includes an inner rod (53) and an outer rod (54) that are slidably sleeved, the upper surface of the top cover (12) close to the edge is circumferentially provided with a plurality of bumps (13) at equal intervals, the top surface of the bump (13) is arc-shaped, the inner rod (53) corresponds to the bump (13) after extending to lift the telescopic gear lever (52), one side of the upper surface of the top cover (12) is provided with an opening (14) for delivering hair samples and ultrapure water, and the bottom of the outer side wall of the treatment barrel (1) is provided with a drainage outlet (15).

### Claim 7 (depends on 6)

The device according to claim 6, wherein a total of 6 groups of hair cutting assemblies (6) are provided, the tail ends of the auxiliary cutting quartz knife (61) and the cutting quartz knife (62) are aligned, the surface of the cutting quartz knife (62) is provided with a plurality of permeable pores (64), the surface of the auxiliary cutting quartz knife (61) is provided with a slot (65), a total of 6 bumps are provided, the auxiliary cutting quartz knife (61) is connected with a clamp groove (44) arranged on the outer wall of the rotating outer cylinder (4) through a fixation block (43), the top and bottom of the fixation block (43) are respectively connected with the top surface and bottom surface of the clamp groove (44) through a first spring rod (45), the two sides of the outer wall of the rotating inner cylinder (5) are provided with limit strips (55), the limit strips (55) are both slidably connected with first limit grooves (46) arranged at the two sides of the inside of the rotating outer cylinder (4) and second limit grooves (33) arranged at the two sides of the inner wall of the sleeve (32), the bottom of the rotating inner cylinder (5) is provided with a weighting block (56), the bottom of the weighting block (56) is connected with the inner bottom of the rotating outer cylinder (4) through a second spring rod (57).

### Claim 8 (depends on 6)

The device according to claim 6, wherein the two sides of the upper surface of the pedestal (2) are symmetrically provided with struts (21), the top of the strut (21) is fixedly connected with the rotating motor (31) through one connection rod (22), the junction between the connection rod (22) and the rotating motor (31) is provided with a damping spacer (34), the telescopic rods (35) are arranged below the two connection rods (22), the bottoms of the two telescopic rods (35) are provided with a limit plate (36) of the inner rod (53) for downward pressing the telescopic gear lever (52), the tail end of the outer rod (54) is provided with a limit hole (58), the tail end of the inner rod (53) is provided with a limit protrusion (59) abutted against the limit hole (58), 3 insulation warehouses (23) are arranged inside the pedestal (2), temperature adjusters (24) are arranged inside the insulation warehouses (23), and the two sides of the upper surface of the pedestal (2) are each provided with an observation beaker (25).

### Claim 9 (depends on 1)

Application of the method according to claim 1, wherein the method for synchronous extraction and analysis of multiple trace emerging organic contaminants in hairs is applied to a study on long-term exposure of multiple organic contaminants in a human body, and the organic contaminants are PBDEs, OPFRs, PFASs, PAEs and mPAEs.