WO2019196233A1 - Method for measuring contents of various endocrine disruptors in estuary sediment - Google Patents

Abstract

Disclosed is a method for measuring the contents of various endocrine disruptors in estuary sediment. Aluminum oxide and diatomaceous earth are employed for a pretreatment of an estuary sediment sample, Soxhlet extraction and a solid-phase extraction method are jointly used for purification to produce an enriched sample, and finally, LC-MS/MS is employed to detect the types and contents of endocrine disruptors in the water sample. The present invention allows the detection of as much as ten types of endocrine disruptors. The detection method of the present invention provides a high extraction efficiency, fast measurement rate, and reliable data analysis, and is applicable in the environmental detection and evaluation of the various endocrine disruptors in estuary sediment.

Description

 Method for determining content of various endocrine disruptors in estuarine sediments

Technical field

The invention relates to the technical field of trace organic matter detection, in particular to a method for determining the content of various endocrine disruptors in estuarine sediments.

Background technique

Endocrine disruptors are exogenous chemicals in the environment that interfere with the normal endocrine system of the organism and cause abnormal effects. Studies have shown that endocrine disruptors can be detected in sediments, water bodies, soils and organisms, and exhibit persistence, toxicity, accumulation and long-distance transmission in the environment. Hundreds of chemicals have been identified as endocrine disruptors.

Estuary sediments are one of the important components of the aquatic ecosystem and are considered to be aggregates of various organic pollutants. As an important host of surface migration materials and atmospheric scattered matter, estuarine sediments are good carriers for studying environmental changes in the past. Endocrine disruptors entering the water body will collect in the bottom sediment by sedimentation, which may detect various interferences.

At present, a variety of methods for detecting endocrine disruptors have been disclosed at home and abroad, but the existing detection methods are only applicable to the aqueous phase, and it is difficult to simultaneously determine various types of endocrine disruptors in complex estuarine sediments. A method for simultaneous enrichment of phenolic, estrogen and androgen-like endocrine disruptors in drinking water is disclosed in Chinese Patent No. CN107121518A, which is filed on Jan. 31, 2009, which is related to the detection of an endocrine disrupting substance in a water environment. The technology can simultaneously detect 11 kinds of endocrine disruptors in three major categories, but the anti-interference ability is poor, and it is only suitable for the detection of endocrine disruptors in drinking water environment.

The detection of endocrine disruptors in sediments requires the extraction of water samples from the matrix. The extraction method affects both the extraction efficiency of the analyte and the selection of the purification and detection techniques of the analyte. Therefore, it is critical to establish techniques for the determination of endocrine disruptors in different matrices. Considering the complexity of estuarine sediment detection, a single extraction method is not sufficient to deal with multiple endocrine disruptors composed of steroid hormones, phenolic compounds and pharmaceutical compounds in sediments. Therefore, there is a need to propose a method for efficiently extracting and determining the content of various endocrine disruptors in estuarine sediments.

Summary of the invention

In view of the deficiencies of the prior art, the technical problem to be solved by the present invention is to provide a method for determining the content of various endocrine disruptors in estuarine sediments.

The technical solution adopted by the present invention is:

The present invention provides a method for determining the content of various endocrine disruptors in estuarine sediments, comprising the steps of:

(1) taking a lyophilized estuarine sediment sample, adding alumina and diatomaceous earth for grinding, sieving, adding an internal standard to obtain a pretreated sample;

(2) taking the pretreated sample into a solvent for Soxhlet extraction, and concentrating the extract obtained by Soxhlet extraction to obtain a concentrated liquid;

(3) adding a diluent to the concentrated solution, and then adding to the activated solid phase column for extraction, then washing the solid phase column with an eluent and drying in a vacuum, followed by elution with an eluent, collecting Eluent, blow dry with nitrogen, add organic solution to volume, mix and filter to obtain sample liquid to be tested;

(4) The standard curve of endocrine disruptors to be tested was drawn by LC-MS/MS to determine the type and content of endocrine disruptors in estuarine sediments in the water samples.

Preferably, in step (1), the mass ratio of alumina: diatomaceous earth: lyophilized estuarine sediment is 1: (1~1.5): (9-11).

Preferably, the internal standard in step (1) is estradiol-D2, ethinyl estradiol-D4 and progesterone-D9.

Preferably, the solvent in the step (2) is a methanol/acetone solution. The methanol/acetone solution used in the present invention refers to a mixed solution of methanol and acetone. Preferably, the solvent in the step (2) is a solution in which methanol and acetone are mixed at a volume ratio of 1:1, that is, methanol/acetone (1:1, v/v).

Further, in the step (2), the mass ratio of the sample to be tested: the solvent is 1: (40 to 50).

Preferably, the eluent in step (3) is an acetonitrile/methyl tert-butyl ether/ammonium hydroxide solution. In the present invention, acetonitrile/methyl tert-butyl ether/ammonium hydroxide refers to a solution formed by mixing acetonitrile, methyl tert-butyl ether and ammonium hydroxide, and preferably the eluent in step (3) is acetonitrile: methyl uncle Butyl ether: A solution of ammonium hydroxide in a volume ratio of 35:5:1, namely acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/v/v).

Preferably, in step (3), the solid phase cartridge is a C18 solid phase extraction cartridge, and the solid phase cartridge is activated by acetonitrile, acetonitrile/methyl tert-butyl ether/ammonium hydroxide, pure water/acetonitrile solution in sequence. . In the present invention, the pure water/acetonitrile solution refers to an aqueous solution of acetonitrile, and preferably acetonitrile, acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/v/v) is used in the step (3). Pure water/acetonitrile (9:1, v/v) activates the solid phase column.

Preferably, in the step (3): the solid phase column is a C18 solid phase extraction cartridge, and the column is activated by acetonitrile, acetonitrile/methyl t-butyl ether/ammonium hydroxide and pure water/acetonitrile solution in sequence. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, the eluent is acetonitrile/methyl tert-butyl ether/ammonium hydroxide solution, and the organic solution is pure water/methanol solution. Preferably, the diluent in the step (3) is pure water/acetonitrile (9:1, v/v), the eluent is pure water/acetonitrile (9:1, v/v), and the vacuum drying time is 5 min. For acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/v/v), the nitrogen blowing temperature is set to 40 ° C when the nitrogen is blown dry, and the organic solution is pure water at constant volume. Methanol (7:3, v/v) was filtered using a 0.22 μm cartridge PTFE filter.

Preferably, the operating parameters of the LC-MS/MS in the step (4) are: the column type is 30 mm×2.1 mm, the Particle Sz. 1.7 μm C18 core-shell reversed phase column, the column temperature is 40 ° C, and the injection is performed. a flow rate of 0.25mL / min, injection volume lOuL, organic mobile phase containing 0.2% NH ₄ OH to a / pure water, the aqueous phase containing 0.2% NH ₄ OH in water / a mixed solution of an organic solution a For acetonitrile/methanol solution, gradient elution procedure: 0 to 3 min, 95% aqueous phase linearly reduced to 5% for 2 min, returned to 95% at 5 min for 5 min; the endocrine disruptor was diclofenac, bisphenol A, at least one of estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol, the mass spectrum is scanned in negative ion mode, the capillary voltage is 4500V, the drying temperature is 600 ° C, and the atomizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, and progesterone, the mass spectrum is scanned in a negative ion mode with a capillary voltage of 5500 V, a drying temperature of 550 ° C, and a nebulizer pressure of 20 psi. In the present invention, A/pure water refers to an aqueous solution of A, and acetonitrile/methanol refers to a mixed solution of acetonitrile and methanol. Preferably, the organic mobile phase in step (4) is A/pure water containing 0.2% NH ₄ OH (95). : 5, v / v) solution, the aqueous phase is a pure water / A (95: 5, v / v) mixed solution containing 0.2% NH ₄ OH, the organic solution A is acetonitrile / methanol (3: 2, v /v) solution, wherein A/pure water (95:5, v/v) refers to A: pure water in a volume ratio of 95:5, pure water / A (95:5, v/v) refers to Pure water: A has a volume ratio of 95:5.

Preferably, the endocrine disrupting agent is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-ethinyl estradiol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, corpus luteum At least one of the ketones.

The invention combines Soxhlet extraction and solid phase extraction to extract endocrine disruptors in estuarine sediments, overcomes the shortcomings of using Soxhlet extraction method and the large amount of reagents, and achieves efficient enrichment and reduction of target substances. The loss of the target; suitable for the determination of a variety of endocrine disruptors in the estuarine sediments, high detection accuracy, reliable results, and practical.

DRAWINGS

1 is a schematic flow chart of the determination of endocrine disruptors in estuarine sediments according to the present invention;

2 is a graph showing measurement results of different Soxhlet extraction times in Example 4;

3 is a graph showing measurement results of different solid phase extraction columns in Example 5.

The implementation, functional features, and advantages of the present invention will be further described in conjunction with the embodiments.

detailed description

It is understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Example 1:

Referring to Figure 1, the present embodiment provides a method for determining the content of various endocrine disruptors in estuarine sediments, including the following steps:

(1) Precipitation of estuarine sediments: Weigh 5g of lyophilized estuarine sediment samples, add 10 ug/L of 10 kinds of target endocrine disruptor mixed standard solution, add alumina and diatomite for grinding, among which alumina : diatomaceous earth: the mass ratio of the lyophilized estuary sediment is 1:1:9, the obtained solid particles are sieved, and the internal standard is added to complete the pretreatment;

(2) Soxhlet extraction: 1 part by mass of the sample pretreated in the step (1) is added to the cellulose tube, and 40 parts by mass of a methanol/acetone (1:1, v/v) solution is added, and Soxhlet extraction is performed for 8 hours. Then, the obtained extract is rotary evaporated to about 1 to 2 mL;

(3) Activation of solid phase cartridge: Take C18 solid phase extraction cartridge and use 5 mL of acetonitrile, acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/v/v) And solid water / acetonitrile (9:1, v / v) solution to activate the solid phase column;

The concentrated liquid obtained by the rotary evaporation in the step (2) is diluted with 15 mL of pure water/acetonitrile (9:1, v/v), and then added to the activated solid phase column in the step (3) for extraction, and then The column was rinsed by adding 10 mL of pure water/acetonitrile (9:1, v/v) and vacuum dried for 5 min, followed by three portions of 5 mL acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/ v/v) elution, collecting the eluent, drying with nitrogen, setting the drying temperature of the nitrogen blowing instrument to 40 ° C, adding organic solution pure water / methanol (7:3, v / v) to volume, using 0.22 μm The tubular PTFE membrane is filtered to obtain a sample liquid to be tested;

(4) Using LC-MS/MS to draw a standard curve of endocrine disruptors in the sample liquid to be tested, thereby determining the type and content of endocrine disruptors in the estuarine sediments of the water sample. The operating parameters of LC-MS/MS are: The column model is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed-phase column, column temperature is 40 ° C, injection flow rate is 0.25 mL / min, injection volume is 10 uL, organic mobile phase is 0.2 % NH ₄ OH to a / water (95: 5, v / v ) solution, the aqueous phase containing 0.2% NH ₄ OH in water / a (95: 5, v / v) mixed solution, the organic solution A is an acetonitrile/methanol (3:2, v/v) solution, gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, and returned to 95% at 5 min for 5 min; When the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol, the mass spectrum is scanned in a negative ion mode, the capillary voltage is 4500 V, and dried. The temperature is 600 ° C, the atomizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, and progesterone, the mass spectrum uses negative ions. Scanning, capillary voltage was 5500V, the drying temperature is 550 deg.] C, an atomizer pressure was 20psi.

The results of the recovery, matrix effect, method detection limit and linearity of the detection method of this example were examined as shown in Table 1.

Table 1 Parameters of the detection method of this embodiment

Compound	Recovery rate %	Matrix effect %	Detection limit (ng/g)	Linearity R 2
Negative ion mode
Diclofenac	91	33	0.35	0.991
Bisphenol A	108	30	0.29	0.994
Estradiol	98	26	0.22	0.992
Estrone	99	7	0.02	0.993
Ethynylestradiol	96	27	0.12	0.993
Octylphenol	87	33	0.30	0.992
Nonylphenol	88	29	0.81	0.990
Positive ion mode
Pultamone	92	18	0.12	0.999
Testosterone	96	12	0.06	0.997
Progesterone	94	−5	0.07	0.999

The results show that the endocrine disruptor to be tested has a good linear relationship under the instrumental conditions of the present embodiment, the linear correlation coefficient R ² >0.99, and the method detection limit of the substance to be detected is low, and the detection limit of the method is 0.02 ~ 0.35 ng / g. The detection method of the invention has the advantages of accurate analysis, high recovery rate and strong anti-interference ability, and the detection limit of the low concentration level ensures the possibility of detecting the trace endocrine disruptors of the estuarine sediment by the analytical method.

Example 2:

This embodiment provides a method for determining the content of various endocrine disruptors in estuarine sediments, including the following steps:

(1) Sediment samples collected from an estuary in South China, sediment samples from cold-dry estuaries, and 5 g of lyophilized estuarine sediment samples were weighed, and 10 standards of endocrine disruptor mixed standard solution were added at a concentration of 100 ug/L. The aluminum and diatomaceous earth are ground, wherein the mass ratio of alumina: diatomaceous earth: lyophilized estuary sediment is 1:1.5:11, the obtained solid particles are sieved, and the internal standard is added to complete the pretreatment;

(2) Soxhlet extraction: 1 part by mass of the sample pretreated in the step (1) is added to the cellulose tube, and 40 parts by mass of a methanol/acetone (1:1, v/v) solution is added, and Soxhlet extraction is performed for 8 hours. Then, the obtained extract is rotary evaporated to about 1 to 2 mL;

(3) Activation of solid phase cartridge: Take C18 solid phase extraction cartridge and use 5 mL of acetonitrile, acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/v/v) And solid water / acetonitrile (9:1, v / v) solution to activate the solid phase column;

The concentrated liquid obtained by the rotary evaporation in the step (2) is diluted with 15 mL of pure water/acetonitrile (9:1, v/v), and then added to the activated solid phase column in the step (3) for extraction, and then The column was rinsed by adding 10 mL of pure water/acetonitrile (9:1, v/v) and vacuum dried for 5 min, followed by three portions of 5 mL acetonitrile/methyl tert-butyl ether/ammonium hydroxide (35:5:1, v/ v/v) elution, collecting the eluent, drying with nitrogen, setting the drying temperature of the nitrogen blowing instrument to 40 ° C, adding organic solution pure water / methanol (7:3, v / v) to volume, using 0.22 μm The tubular PTFE membrane is filtered to obtain a sample liquid to be tested;

(4) Using LC-MS/MS to draw a standard curve of endocrine disruptors in the sample liquid to be tested, thereby determining the type and content of endocrine disruptors in the estuarine sediments of the water sample. The operating parameters of LC-MS/MS are: The column model is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed-phase column, column temperature is 40 ° C, injection flow rate is 0.25 mL / min, injection volume is 10 uL, organic mobile phase is 0.2 % NH ₄ OH to a / water (95: 5, v / v ) solution, the aqueous phase containing 0.2% NH ₄ OH in water / a (95: 5, v / v) mixed solution, the organic solution A is an acetonitrile/methanol (3:2, v/v) solution, gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, and returned to 95% at 5 min for 5 min; When the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol, the mass spectrum is scanned in a negative ion mode, the capillary voltage is 4500V, and dried. The temperature is 600 ° C, the atomizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, and progesterone, the mass spectrum uses negative ions. Scanning, capillary voltage was 5500V, the drying temperature is 550 deg.] C, an atomizer pressure was 20psi. The results of analysis and analysis by the above detection steps are shown in Table 2.

Table 2 Test results of sediments in a estuary in South China

Compound	Concentration (ng/g)	Compound	Concentration (ng/g)
Diclofenac	not detected	Octylphenol	not detected
Bisphenol A	93.32	Nonylphenol	332.84
Estradiol	105.24	Pultamone	not detected
Estrone	74.16	Testosterone	3.84
Ethynylestradiol	45.27	Progesterone	1.53

The experimental results show that the detection method of the invention has been successfully applied to the detection of endocrine disruptors in sediments of an estuary in South China.

Comparative Example 1: Comparative Example 1 was the same as Example 1, except that step (3) was removed, and then the concentrate obtained by rotary evaporation in the step (2) was added to pure water/methanol (7:3, v/v). Constant volume, mix the filter membrane to obtain the sample liquid to be tested, and then proceed to step (4).

Comparative Example 2: Comparative Example 1 was the same as Example 1, except that the step (2) was specifically: adding the sample pretreated in the step (1) to a methanol/acetone (1:1, v/v) solution for ultrasonication. After 30 min, the supernatant was obtained by centrifugation and rotary evaporated to about 1 to 2 mL.

The endocrine disruptors in the estuarine sediment were measured by the detection methods in Example 1 and Comparative Example 1-2, and the results are shown in Table 3.

Table 3 Measurement results of the detection methods of Example 1 and Comparative Example 1-2

Compound	Example 1 Recovery %	Comparative Example 1 Recovery %	Comparative Example 2 Recovery %
Diclofenac	91	78	82
Bisphenol A	108	90	116
Estradiol	98	86	87
Estrone	99	87	93
Ethynylestradiol	96	89	85
Octylphenol	87	83	81
Nonylphenol	88	81	75
Pultamone	92	74	106
Testosterone	96	79	72
Progesterone	94	118	91

The experimental results show that the recovery rate of the test substance is significantly better than that of Comparative Example 1 and Comparative Example 2 when using the detection method of the present invention, indicating that the combined effect of Soxhlet extraction and solid phase extraction is significantly better than the separate use of the two. .

Example 3: Soxhlet extraction time optimization

In order to fully utilize the time and achieve efficient extraction of various endocrine disruptors, the present invention optimizes the Soxhlet extraction efficiency and investigates the effects of different extraction times (4h, 6h, 8h and 10h) on the recovery rate. The results are shown in Figure 2. . The experimental results show that with the increase of Soxhlet extraction time, the recovery rate of target endocrine disruptors is continuously improved, when the extraction time is 8 At h, the recovery rate of each component slows down and basically tends to be constant, so the Soxhlet extraction time is 8 h, which ensures high recovery rate and saves time.

Example 4: Optimization of solid phase extraction column

Different extraction columns have different enrichment effects on different types of endocrine disruptors. Since the present invention relates to various types of endocrine disruptors, in order to achieve efficient extraction and enrichment of various endocrine disruptors, the present invention compares three commonly used solids according to experience. The effect of the selection of phase extraction columns (HLB, MCX and C18) on the enrichment efficiency of the target is shown in Figure 3. The results showed that when C18 column was used for extraction, the enrichment effect was the best, and the effective enrichment of many types of endocrine disruptors could be achieved.

The above are only the preferred embodiments of the present invention, and are not intended to limit the scope of the invention, and the equivalent structure or equivalent process transformations made by the description of the present invention and the drawings are directly or indirectly applied to other related technical fields. The same is included in the scope of patent protection of the present invention.

Claims (28)

1.  A method for determining a plurality of endocrine disrupting substances in a estuarine sediment, characterized by comprising the steps of:

   (1) taking a lyophilized estuarine sediment sample, adding alumina and diatomaceous earth for grinding, sieving, adding an internal standard to obtain a pretreated sample;

   (2) taking the pretreated sample into a solvent for Soxhlet extraction, and concentrating the extract obtained by Soxhlet extraction to obtain a concentrated liquid;

   (3) adding a diluent to the concentrated solution, and then adding to the activated solid phase column for extraction, then washing the solid phase column with an eluent and drying in a vacuum, followed by elution with an eluent, collecting Eluent, dried by nitrogen, Adding organic solution to volume, mixing and filtering to obtain sample liquid to be tested;

   (4) Using LC-MS/MS A standard curve of endocrine disruptors in the sample liquid to be tested is drawn to determine the type and content of endocrine disruptors in the estuarine sediments of the water sample.
2. According to claim 1 The method for determining a plurality of endocrine disrupting substances in a estuarine sediment, characterized in that the mass ratio of alumina: diatomaceous earth: lyophilized estuary sediment in step (1) is 1: (1~1.5): (9~11).
3. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 1, wherein the internal standard in the step (1) is estradiol-D2, ethinyl estradiol-D4 and progesterone- D9.
4. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 1, wherein the solvent in the step (2) is a methanol/acetone solution.
5. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 4, wherein the mass ratio of the sample to be tested in the step (2) is 1: (40 to 50).
6. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 1, wherein the eluent in the step (3) is an acetonitrile/methyl tert-butyl ether/ammonium hydroxide solution.
7. The method for determining a plurality of endocrine disrupting substances in a estuary sediment according to claim 1, wherein in the step (3), the solid phase column is a C18 solid phase extraction column, and acetonitrile, acetonitrile/A is sequentially used. The solid phase cartridge was activated by a solution of tert-butyl ether/ammonium hydroxide and pure water/acetonitrile.
8. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 1, wherein in the step (3): the solid phase small column is a C18 solid phase extraction column, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
9. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 2, wherein in the step (3): the solid phase small column is a C18 solid phase extraction cartridge, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
10. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 3, wherein in the step (3): the solid phase small column is a C18 solid phase extraction cartridge, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
11. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 4, wherein in the step (3): the solid phase small column is a C18 solid phase extraction cartridge, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
12. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 5, wherein in the step (3): the solid phase small column is a C18 solid phase extraction column, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
13. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 6, wherein in the step (3): the solid phase small column is a C18 solid phase extraction cartridge, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
14. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 7, wherein in the step (3): the solid phase column is a C18 solid phase extraction cartridge, and acetonitrile and acetonitrile are sequentially used. The column is activated by methyl tert-butyl ether/ammonium hydroxide and pure water/acetonitrile solution. The diluent is pure water/acetonitrile solution, the eluent is pure water/acetonitrile, and the eluent is acetonitrile/methyl tert-butyl. The ether/ammonium hydroxide solution, the organic solution is a pure water/methanol solution.
15. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 1, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
16. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 2, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
17. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 3, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
18. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 4, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
19. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 5, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
20. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 6, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
21. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 7, wherein the operating parameter of the LC-MS/MS in the step (4) is: the column type is 30 mm × 2.1 mm, Particle Sz. 1.7 μm C18 core-shell reversed phase column with a column temperature of 40 ° C, a sample flow rate of 0.25 mL/min, an injection volume of 10 uL, and an organic mobile phase of A/pure aqueous solution containing 0.2% NH ₄ OH. The aqueous phase is a pure water/A mixed solution containing 0.2% NH ₄ OH, the A is an acetonitrile/methanol solution, and a gradient elution procedure: 0 to 3 min, 95% of the aqueous phase is linearly reduced to 5% for 2 min, at Return to 95% at 5 min for 5 min; the endocrine disruptor is at least one of diclofenac, bisphenol A, estradiol, estrone, ethinyl estradiol, octylphenol, nonylphenol. Negative ion mode scanning, capillary voltage is 4500V, drying temperature is 600 ° C, nebulizer pressure is 20 psi; when the endocrine disruptor is at least one of primidone, testosterone, progesterone, mass spectrometry is scanned by negative ion mode, capillary The voltage is 5500V, the drying temperature is 550 ° C, and the atomizer pressure is 20 psi.
22. The method according to claim 1, wherein the endocrine disruptor is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.
23. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 2, wherein the endocrine disrupting agent is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.
24. The method according to claim 3, wherein the endocrine disruptor is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.
25. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 4, wherein the endocrine disrupting agent is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.
26. The method for determining a plurality of endocrine disrupting substances in a estuarine sediment according to claim 5, wherein the endocrine disrupting agent is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.
27. The method according to claim 6, wherein the endocrine disruptor is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.
28. The method according to claim 7, wherein the endocrine disruptor is diclofenac, bisphenol A, 17β-estradiol, estrone, 17α-acetylene female At least one of a diol, 4-octylphenol, 4-nonylphenol, primidone, testosterone, and progesterone.