Application of solid phase extraction nanofiber material in detection of hormone drugs in aquatic products

Abstract

The application discloses a kind of solid phase extraction nanofiber material detection hormone drugs in aquatic products, and relates to drug detection technical field.The specific preparation method includes: first, an activated solid phase extraction column is prepared, and 2D-MOF is prepared using ligand, metal source and alkali;Then polymer and solvent, or 2D-MOF, polymer and solvent are mixed to obtain electrospinning solution, electrospinning solution is electrospun to obtain electrospinning film, and drying is nanofiber;Activated solid phase extraction column is made using nanofiber.Hormone drugs in the extract of aquatic product sample are enriched and purified using activated solid phase extraction column, and 18 kinds of hormone drugs in aquatic products are detected based on ultra-high performance liquid chromatography-tandem mass spectrometry.The activated solid phase extraction column of the application has good enrichment recovery effect on hormone drugs, and the detection method is accurate and sensitive.

Description

Technical Field

[0001] This invention relates to the field of drug detection technology, specifically to the application of solid-phase extraction nanofiber materials in the detection of hormonal drugs in aquatic products. Background Technology

[0002] The frequent misuse of pesticides and veterinary drugs in aquaculture seriously affects food safety. Hormones are chemical substances produced by cells, glands, or organs in the body that can affect the activity of other cells. They mainly include sex hormones, progestins, and glucocorticoids. The coordinated action of various hormones is essential for maintaining the body's metabolism and function. Many hormone preparations and synthetic products have important uses in medicine, animal husbandry, and fisheries. With the development of high-density aquaculture technology, drugs are widely used for the treatment and prevention of diseases in farmed animals. They are also added to animal feed and drinking water for non-therapeutic purposes, mainly to promote growth, improve feed conversion rate, and enhance product quality. In aquaculture, hormone drugs control the reproductive activity of fish through "hormonal regulation," primarily using exogenous hormones for artificial reproduction, artificial sex control, and accelerating the breeding of broodstock in fish and other farmed aquatic animals. Generally, hormone drugs have a relatively stable structure and are not easily degraded. Excessive and inappropriate use can lead to hormone drug residues in animals, which can then enter the human body through the food chain, harming health and causing diseases such as obesity and hypertension.

[0003] Currently, methods for determining various hormonal drugs in food mainly include liquid chromatography / triple quadrupole tandem mass spectrometry, liquid chromatography-quadrupole / time-of-flight mass spectrometry, gas chromatography-mass spectrometry, and capillary electrophoresis. High-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) is a commonly used method for trace analysis due to its high sensitivity and selectivity; however, sample purification is required to reduce matrix interference, and traditional solid-phase extraction pretreatment methods are time-consuming and labor-intensive. Therefore, the pretreatment of aquatic samples is a major challenge, as the rich and complex matrix of aquatic samples easily leads to matrix effects and inaccurate direct determination of trace compounds. Therefore, there is an urgent need for a simple, rapid, and effective technique for detecting hormonal drugs. Summary of the Invention

[0004] The purpose of this invention is to provide an application of solid-phase extraction nanofiber materials for the detection of hormonal drugs in aquatic products, so as to solve the problems of strong matrix interference and time-consuming and labor-intensive methods in existing detection methods.

[0005] To solve the above-mentioned technical problems, the present invention specifically provides the following technical solution:

[0006] This invention discloses a method for preparing an activated solid-phase extraction column, comprising:

[0007] S1: The solvent is mixed to obtain a mixture, then the ligand and metal source are added and stirred, then the alkali is added and ultrasonic treatment is performed, and finally the 2D-MOF is obtained after washing.

[0008] S2: The polymer is mixed with a solvent, or 2D-MOF, polymer and solvent are mixed to obtain an electrospinning solution. The electrospinning solution is electrospinned to obtain an electrospinning film, which is then dried to obtain nanofibers. The mass ratio of the amount of 2D-MOF to polymer used is 1:1-5.

[0009] S3: First, the tablet is fed to the lower end of the solid phase extraction column, then nanofibers are placed in, and then the tablet is pressed to obtain a solid phase extraction column. The nanofibers are activated by flowing an activator through the column to obtain an activated solid phase extraction column.

[0010] Preferably, the solvent in S1 is N,N-dimethylformamide, ethanol and water, the volume ratio of N,N-dimethylformamide to ethanol is 1:0.03-0.1, the volume ratio of N,N-dimethylformamide to water is 1:0.03-0.1; the ligand is terephthalic acid, and the volume ratio of terephthalic acid to N,N-dimethylformamide is 1 mmol:40-50 ml.

[0011] Preferably, the metal source in S1 is at least one of MnCl2·4H2O, CoCl2·6H2O and NiCl2·6H2O, and the molar ratio of the amount of ligand to metal source is 1:0.5-1.5.

[0012] Preferably, the base in S1 is triethylamine, and the ratio of ligand to base is 1 mmol: 0.8-1.5 ml.

[0013] Preferably, the polymer in S2 is at least one selected from polystyrene, polyacrylonitrile, polyvinyl alcohol, and modified polystyrene; the modified polystyrene is prepared from 4-chlorobutyryl chloride, polystyrene, and 2,2'-(propane-2,2-dimethylbis(sulfonamide dimethyl))diethylamine. Nanofibers prepared using modified polystyrene exhibit good adsorption properties, enabling the extraction and enrichment of target hormone drugs from complex media, thereby achieving high recovery rates of target hormone drugs and reducing the influence of matrix effects during subsequent ultra-high performance liquid chromatography-tandem mass spectrometry detection.

[0014] Preferably, the solvent in S2 is N,N-dimethylformamide; the ratio of polymer to solvent is 1g:10-20ml.

[0015] Preferably, the activator in S3 is methanol and water, which flow sequentially through the nanofibers inside the column. The ratio of nanofibers to methanol is 1g:20-30ml, and the ratio of nanofibers to water is 1g:40-60ml.

[0016] Preferably, the stirring time in S1 is 20-40 min, and the ultrasonic treatment time is 6-10 h.

[0017] Preferably, in S2, the curing distance of electrospinning is 15-25cm, the voltage is 15-25kv, and the injection speed is 40-50uL / min.

[0018] The present invention also discloses the application of any of the above-mentioned activated solid-phase extraction columns in the preparation of purified aquatic samples.

[0019] This invention discloses a method for preparing an activated solid-phase extraction column, comprising:

[0020] S1: Mix the solvent to obtain a mixture, then add the ligand and metal source and stir for 20-40 min, then add alkali and sonicate for 6-10 h, and finally wash to obtain 2D-MOF;

[0021] S2: The polymer is mixed with a solvent, or 2D-MOF and polymer are mixed with a solvent to obtain an electrospinning solution. The electrospinning solution is electrospinned at a curing distance of 15-25cm, a voltage of 15-25kv and an injection speed of 40-50uL / min to obtain an electrospinned film. Finally, it is dried to obtain nanofibers.

[0022] S3: First, the tablet is fed to the lower end of the solid phase extraction column, then nanofibers are placed in, and then the tablet is pressed to obtain a solid phase extraction column. The nanofibers are activated by flowing an activator through the column to obtain an activated solid phase extraction column.

[0023] Preferably, the solvent in S1 is N,N-dimethylformamide, ethanol and water.

[0024] More preferably, the volume ratio of N,N-dimethylformamide to ethanol is 1:0.03-0.1.

[0025] More preferably, the volume ratio of N,N-dimethylformamide to water is 1:0.03-0.1.

[0026] Preferably, the ligand in S1 is terephthalic acid.

[0027] More preferably, the ratio of terephthalic acid to N,N-dimethylformamide is 1 mmol: 40-50 ml.

[0028] Preferably, the metal source in S1 is at least one of MnCl2·4H2O, CoCl2·6H2O, and NiCl2·6H2O.

[0029] Preferably, the molar ratio of the amount of ligand to metal source used in S1 is 1:0.5-1.5.

[0030] Preferably, the base in S1 is triethylamine.

[0031] Preferably, the ratio of ligand to base in S1 is 1 mmol: 0.8-1.5 ml.

[0032] Preferably, the solvent used for washing in S1 is ethanol and water, and the washing is performed sequentially by ethanol and water.

[0033] Preferably, the polymer in S2 is at least one of polystyrene, polyacrylonitrile, polyvinyl alcohol, and modified polystyrene.

[0034] Preferably, the solvent in S2 is N,N-dimethylformamide.

[0035] Preferably, the ratio of polymer to solvent in S2 is 1g:10-20ml.

[0036] Preferably, the mass ratio of 2D-MOF to polymer in S2 is 1:1-5.

[0037] Preferably, the activator in S3 is methanol and water, and methanol and water flow sequentially through the nanofibers inside the column.

[0038] More preferably, the ratio of nanofibers to methanol is 1g:20-30ml.

[0039] More preferably, the ratio of nanofibers to water is 1g:40-60ml.

[0040] This invention discloses a method for preparing modified polystyrene, specifically as follows:

[0041] Aluminum chloride, 4-chlorobutyryl chloride, and chloroform were mixed. This mixture was then added dropwise to a polystyrene solution and reacted at 25-35°C for 2-5 hours. After the reaction was complete, the mixture was washed, and then ethanol was added to separate the precipitate, which was then dried to obtain an intermediate. N,N-dimethylformamide and 2,2'-(propane-2,2-diylbis(sulfonamide diel))diethylamine were added to the intermediate, and the mixture was reacted at 50-60°C for 2-5 hours. Finally, the mixture was washed and dried to obtain modified polystyrene.

[0042] Preferably, the mass ratio of aluminum chloride to 4-chlorobutyryl chloride used is 1:0.8-1.2.

[0043] Preferably, the ratio of aluminum chloride to chloroform is 1g:10-20ml.

[0044] Preferably, the polystyrene solution is composed of polystyrene and N,N-dimethylformamide.

[0045] More preferably, the ratio of polystyrene to N,N-dimethylformamide is 1g:8-15ml.

[0046] Preferably, the mass ratio of the polystyrene solution to the mixed solution is 1:0.001-0.003.

[0047] Preferably, the ratio of the intermediate to N,N-dimethylformamide is 1g:8-15ml.

[0048] Preferably, the mass ratio of the intermediate to 2,2'-(propane-2,2-dimethylbis(sulfonamide dimethyl))diethylamine is 1:0.002-0.008.

[0049] Preferably, the solvent used for washing during the preparation of the intermediate is hydrochloric acid solution and water, and the hydrochloric acid solution and water are used for washing in sequence. The hydrochloric acid solution is composed of hydrochloric acid and water, and the ratio of hydrochloric acid to water is 1g:8-15ml.

[0050] Preferably, the solvent used for washing during the preparation of modified polystyrene is ethanol.

[0051] Preferably, in the preparation of nanofibers, in addition to using modified polystyrene, poly(1,4-cyclohexanedimethyl adipate) can also be used. The synergistic use of modified polystyrene and poly(1,4-cyclohexanedimethyl adipate) prepares nanofibers, which makes the nanofibers contain a variety of functional groups, have good adsorption properties for target hormone drugs, good recovery rate after adsorption, and good matrix effect removal effect, effectively improving the accuracy of subsequent detection.

[0052] More preferably, the mass ratio of polystyrene to poly(1,4-cyclohexanedimethyl adipate) is 1:0.5-1.2.

[0053] Compared with the prior art, the present invention has the following advantages:

[0054] This invention proposes an application of solid-phase extraction (SPE) nanofiber materials for detecting hormonal drugs in aquatic products. First, an activated SPE column is prepared, and 2D-MOF is obtained using ligands, a metal source, and an alkali. Then, an electrospinning solution is prepared by mixing a polymer with a solvent, or by mixing 2D-MOF, a polymer, and a solvent. This electrospinning solution is then electrospinned to obtain an electrospinned membrane, which is dried to obtain nanofibers. The activated SPE column is then used to fabricate the activated SPE column. The activated SPE column is used to enrich and purify hormonal drugs in the extract of aquatic product samples, and 18 hormonal drugs in the aquatic products are detected based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS). The activated SPE column of this invention exhibits good enrichment and recovery effects for hormonal drugs, and the detection method is accurate and sensitive. Attached Figure Description

[0055] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.

[0056] Figure 1 SEM image of the nanofibers prepared in Example 1;

[0057] Figure 2 Here is a SEM image of the nanofibers prepared in Example 4;

[0058] Figure 3 Here is a SEM image of the nanofibers prepared in Example 5;

[0059] Figure 4 Here is a SEM image of the nanofibers prepared in Example 6;

[0060] Figure 5 Ion chromatograms of 18 hormone drugs;

[0061] Figure 6 The results show the recovery rates of 18 types of hormone drugs. Detailed Implementation

[0062] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0063] The concepts involved in this application will first be described with reference to the accompanying drawings. It should be noted that the following descriptions of various concepts are only for the purpose of making the content of this application easier to understand and do not constitute a limitation on the scope of protection of this application; furthermore, the embodiments and features in the embodiments of this application can be combined with each other unless otherwise specified. This application will now be described in detail with reference to the accompanying drawings and embodiments.

[0064] The specific meanings of the abbreviations used in the specification and claims are as follows:

[0065] Example 1:

[0066] Preparation of nanofibers: Polystyrene and N,N-dimethylformamide were mixed and stirred for 24 h to obtain an electrospinning solution. The electrospinning solution was then placed in an electrospinning apparatus and sprayed at a curing distance of 20 cm, a voltage of 20 kV, and an injection speed of 45 μL / min to obtain an electrospinned film. The electrospinned film was dried to obtain nanofibers. The ratio of polystyrene to N,N-dimethylformamide was 1 g: 7 ml.

[0067] Preparation of the activated solid-phase extraction column: First, a tablet was fed to the lower end of the solid-phase extraction column using a solid metal cylinder. Then, nanofibers were placed inside, followed by another tablet, and the column was compressed using the solid metal cylinder to obtain the solid-phase extraction column. The nanofibers inside the column were then activated by sequentially passing methanol and water through them to obtain the activated solid-phase extraction column. The ratio of nanofibers to methanol was 1 g: 25 ml, and the ratio of nanofibers to water was 1 g: 50 ml.

[0068] Example 2:

[0069] Preparation of nanofibers: The preparation of nanofibers in this example differs from that in Example 1 in that polystyrene is replaced with polyacrylonitrile, while other conditions and parameters are the same as in Example 1.

[0070] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 1, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 1.

[0071] Example 3:

[0072] Preparation of nanofibers: The preparation of nanofibers in this example differs from that in Example 1 in that polystyrene is replaced with polyvinyl alcohol, while other conditions and parameters are the same as in Example 1.

[0073] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 1, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 1.

[0074] Example 4:

[0075] Preparation of 2D-MOF: First, N,N-dimethylformamide, ethanol, and water were mixed to obtain a mixture. Then, terephthalic acid and MnCl2·4H2O were added, and the mixture was stirred for 30 min. After stirring, triethylamine was added, and the mixture was sonicated for 8 h. Finally, the mixture was washed with ethanol and water sequentially and dried to obtain 2D-MOF. The volume ratio of N,N-dimethylformamide to ethanol in the mixture was 1:0.0625, the volume ratio of N,N-dimethylformamide to water was 1:0.0625, the volume ratio of terephthalic acid to N,N-dimethylformamide was 1 mmol:42.67 ml, the molar ratio of terephthalic acid to MnCl2·4H2O was 1:1, and the volume ratio of terephthalic acid to triethylamine was 1 mmol:1.07 ml.

[0076] Preparation of nanofibers: 2D-MOF, polystyrene, and N,N-dimethylformamide were mixed and stirred for 24 hours to obtain an electrospinning solution. The electrospinning solution was then placed in an electrospinning apparatus and sprayed at a curing distance of 20 cm, a voltage of 20 kV, and an injection speed of 45 μL / min to obtain an electrospinned film. The electrospinned film was dried to obtain nanofibers. The mass ratio of 2D-MOF to polystyrene was 1:1.5, and the ratio of polystyrene to N,N-dimethylformamide was 1 g:13.33 ml.

[0077] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 1, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 1.

[0078] Example 5:

[0079] Preparation of 2D-MOF: The preparation of 2D-MOF in this example differs from that in Example 4 in that MnCl2·4H2O is replaced with CoCl2·6H2O, while other conditions and parameters are the same as in Example 4.

[0080] Preparation of nanofibers: The preparation of nanofibers in this embodiment differs from that in Example 4 in that the 2D-MOF used is the 2D-MOF prepared in this embodiment, while other conditions and parameters are the same as in Example 4.

[0081] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 4, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 4.

[0082] Example 6:

[0083] Preparation of 2D-MOF: The preparation of 2D-MOF in this example differs from that in Example 4 in that MnCl2·4H2O is replaced with NiCl2·6H2O, while other conditions and parameters are the same as in Example 4.

[0084] Preparation of nanofibers: The preparation of nanofibers in this embodiment differs from that in Example 4 in that the 2D-MOF used is the 2D-MOF prepared in this embodiment, while other conditions and parameters are the same as in Example 4.

[0085] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 4, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 4.

[0086] Example 7:

[0087] The preparation of 2D-MOF is the same as in Example 6.

[0088] Preparation of modified polystyrene: Aluminum chloride, 4-chlorobutyryl chloride, and chloroform were mixed to obtain a mixed solution. This mixed solution was then added dropwise to a polystyrene solution, and the reaction was carried out at 30°C for 3 hours. After the reaction, the mixture was washed successively with hydrochloric acid and water, and then ethanol was added to separate the precipitate, which was then dried to obtain an intermediate. N,N-dimethylformamide and 2,2'-(propane-2,2-diylbis(sulfonamide diel))diethylamine were added to the intermediate, and the reaction was carried out at 55°C for 3 hours. After the reaction, the mixture was washed with ethanol and dried to obtain modified polystyrene. The mass ratio of aluminum chloride to 4-chlorobutyryl chloride used is 1:1.06; the mass ratio of aluminum chloride to chloroform is 1g:15ml; the polystyrene solution is composed of polystyrene and N,N-dimethylformamide, with a mass ratio of polystyrene to N,N-dimethylformamide of 1g:10ml; the mass ratio of polystyrene solution to mixed solution is 1:0.0015; the hydrochloric acid solution is composed of hydrochloric acid and water, with a mass ratio of hydrochloric acid to water of 1g:10ml; the mass ratio of intermediate to N,N-dimethylformamide is 1g:10ml; and the mass ratio of intermediate to 2,2'-(propane-2,2-dimethylbis(sulfonamide dimethyl))diethylamine is 1:0.005.

[0089] Preparation of nanofibers: The preparation of nanofibers in this embodiment differs from that in Example 6 in that polystyrene is replaced with modified polystyrene, while other conditions and parameters are the same as in Example 6.

[0090] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 6, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 6.

[0091] Example 8:

[0092] The preparation of 2D-MOF is the same as in Example 6.

[0093] The preparation of modified polystyrene is the same as in Example 7.

[0094] Preparation of nanofibers: The preparation of nanofibers in this example differs from that in Example 7 in that the mass ratio of 2D-MOF to modified polystyrene is 1:2, while other conditions and parameters are the same as in Example 7.

[0095] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 7, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 7.

[0096] Example 9:

[0097] The preparation of 2D-MOF is the same as in Example 6.

[0098] The preparation of modified polystyrene is the same as in Example 7.

[0099] Preparation of nanofibers: 2D-MOF, modified polystyrene, poly(1,4-cyclohexanedimethyl adipate), and N,N-dimethylformamide were mixed and stirred for 24 h to obtain an electrospinning solution. The electrospinning solution was then placed in an electrospinning apparatus and sprayed at a curing distance of 20 cm, a voltage of 20 kV, and an injection speed of 45 μL / min to obtain an electrospinned film. The electrospinned film was dried to obtain nanofibers. The mass ratio of 2D-MOF to modified polystyrene was 1:1.5, the mass ratio of modified polystyrene to N,N-dimethylformamide was 1 g:13.33 ml, and the mass ratio of modified polystyrene to poly(1,4-cyclohexanedimethyl adipate) was 1:0.6.

[0100] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 7, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 7.

[0101] Example 10:

[0102] The preparation of 2D-MOF is the same as in Example 6.

[0103] The preparation of modified polystyrene is the same as in Example 7.

[0104] Preparation of nanofibers: The preparation of nanofibers in this example differs from that in Example 9 in that the mass ratio of modified polystyrene to poly(1,4-cyclohexanedimethyl adipate) is 1:1, while other conditions and parameters are the same as in Example 9.

[0105] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this embodiment is different from that in Example 9, except that the nanofibers are prepared in this embodiment, while other conditions and parameters are the same as in Example 9.

[0106] Comparative Example 1:

[0107] The preparation of 2D-MOF is the same as in Example 6.

[0108] The preparation of modified polystyrene is the same as in Example 7.

[0109] Preparation of nanofibers: The preparation of nanofibers in this example differs from that in Example 7 in that the mass ratio of 2D-MOF to modified polystyrene is 1:0.2, while other conditions and parameters are the same as in Example 7.

[0110] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this comparative example is different from that in Example 7, except that the nanofibers are the nanofibers prepared in this example, and other conditions and parameters are the same as in Example 7.

[0111] Comparative Example 2:

[0112] The preparation of 2D-MOF is the same as in Example 6.

[0113] The preparation of modified polystyrene is the same as in Example 7.

[0114] Preparation of nanofibers: The preparation of nanofibers in this comparative example differs from that in Example 9 in that modified polystyrene was not used, while other conditions and parameters are the same as in Example 9.

[0115] Preparation of activated solid phase extraction column: The preparation of activated solid phase extraction column in this comparative example is different from that in Example 9, except that the nanofibers are the nanofibers prepared in this example, and the other conditions and parameters are the same as in Example 9.

[0116] Experimental Example 1:

[0117] Nanofiber morphology characterization: The nanofibers prepared in Examples 1 and 4-6 were examined by scanning electron microscopy (SEM) to obtain SEM images. The SEM image of the nanofibers prepared in Example 1 is shown below. Figure 1 As shown, the SEM image of the nanofibers prepared in Example 4 is as follows. Figure 2 As shown, the SEM image of the nanofibers prepared in Example 5 is as follows. Figure 3 As shown, the SEM image of the nanofibers prepared in Example 6 is as follows. Figure 4 As shown, the nanofibers exhibit a beaded structure and good hydrophobicity.

[0118] Experimental Example 2:

[0119] Preparation of hormone drug test samples: Eighteen hormone drug standards found in aquatic products were dissolved in methanol to obtain a mixed standard intermediate solution. This mixed standard intermediate solution was then dissolved in a methanol-acetonitrile aqueous solution to obtain a mixed standard solution. The mixed standard solution was passed through an activated solid-phase extraction (SPE) column, followed by elution with acidified methanol. The eluent was collected as the hormone drug test sample. The concentration of the 18 hormone drug standards in the mixed standard intermediate solution was 1 µg / mL, and the concentration of the 18 hormone drug standards in the mixed standard solution was 100 ng / mL. The acidified methanol consisted of acetic acid and methanol, with a volume ratio of 1:99. The activated SPE column was the one prepared in Examples 1-3.

[0120] The recovery rate of hormone drugs after adsorption by activated solid-phase extraction column was determined by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) to detect hormone drugs in the test samples, and the recovery rates of 18 hormone standards were calculated to compare the adsorption efficiency of activated solid-phase extraction column.

[0121] The detection conditions for ultra-high performance liquid chromatography are as follows:

[0122] The chromatographic column was a ZORBAX Eclipse Plus C18 column (2.1 × 100 mm, 1.8-Micron); the column temperature was 325℃; the flow rate was 0.3 mL / min; the injection volume was 5 μL; mobile phase A consisted of water, ammonium acetate, and formic acid, with a volume ratio of 1:0.001:0.00; mobile phase B was methanol; and the gradient elution program was shown in Table 1.

[0123] Table 1 Gradient Elution Table

[0124]

[0125] The mass spectrometry detection conditions are as follows:

[0126] The ion source was a dual-jet electrospray ionization source (Dual AJF ESI), with positive ion mode and multiple reaction monitoring (MRM) mode. The nebulizer pressure was 275 kPa; the gas temperature was 350 °C; the gas flow rate was 5 L / min; the sheath gas temperature was 300 °C; the sheath gas flow rate was 11 L / min; and the capillary voltage was 4000 V. The mass spectrometry parameters of 18 hormone drugs are shown in Table 2, and the ion chromatograms are shown below. Figure 5 As shown.

[0127] Table 2 Mass spectrometry parameters of 18 hormone drugs

[0128]

[0129] After using the activated solid-phase extraction columns prepared in Examples 1-3 to adsorb and purify hormone drugs, the recovery rate of hormone drugs was detected as follows: Figure 6 As shown, the 18 hormone drugs showed the best recovery rate after adsorption using the activated solid-phase extraction column prepared in Example 1, with recovery rates ranging from 75.7% to 114.7%.

[0130] Experimental Example 3:

[0131] The recovery rate of megestrol acetate after adsorption of hormone drugs by the activated solid-phase extraction column was determined using the same method as in Example 2, except that the activated solid-phase extraction column was the one prepared in Examples 1-10 and Comparative Examples 1-2.

[0132] Table 3 Results of recovery rate of medroxyprogesterone acetate (a hormone drug)

[0133]

[0134] The recovery rates of megestrol acetate (MEA) after adsorption and purification of hormonal drugs using the activated solid-phase extraction columns prepared in Examples 1-9 and Comparative Examples 1-2 are shown in Table 3. Compared with Examples 2-3, the activated solid-phase extraction column prepared in Example 1 showed the best recovery rate and adsorption efficiency. Compared with Examples 4-6, Example 1 demonstrates that the preparation of 2D-MOF using NiCl2·4H2O, and its subsequent use in the preparation of nanofibers and activated solid-phase extraction columns, can improve the recovery rate of the prepared activated solid-phase extraction column, achieving the best recovery effect. Compared with Example 7, Example 6 demonstrates that the use of modified polystyrene can... To further improve the effect of the prepared activated solid-phase extraction column and increase the recovery rate after adsorption; Example 7, compared with Example 8, shows that increasing the amount of modified polystyrene within a certain range can improve the recovery rate after adsorption by the prepared activated solid-phase extraction column, that is, improve the adsorption efficiency; Example 7, compared with Example 9, shows that using poly(1,4-cyclohexanedimethyl adipate) to prepare nanofibers based on modified polystyrene and using them for subsequent preparation of activated solid-phase extraction columns can improve the recovery rate after adsorption by the prepared activated solid-phase extraction column; Example 9, compared with Example 10, shows that poly(1,4-cyclohexanedimethyl adipate) Increasing the amount of modified polystyrene used within a certain range can further improve the recovery rate of the activated solid-phase extraction column after adsorption. Compared with Comparative Example 1, Example 7 shows that the amount of modified polystyrene used needs to be within a suitable range. Too little will reduce the recovery rate of the activated solid-phase extraction column after adsorption. Compared with Comparative Example 2, Example 9 shows that modified polystyrene and poly(1,4-cyclohexanedimethyl adipate) need to be used together. Using poly(1,4-cyclohexanedimethyl adipate) alone has no significant effect on improving the recovery rate of the activated solid-phase extraction column after adsorption.

[0135] Experiment Example 4:

[0136] Preparation of blank sample matrix: Add ethyl acetate to the blank aquatic sample, vortex for 15 min, then centrifuge to collect the supernatant, dry it with nitrogen, then add methanol-acetonitrile aqueous solution to redissolve it, filter it with a 0.22 μm organic filter membrane, pass it through an activated solid-phase extraction column, then elute the solid-phase extraction column with acidified methanol, collect the eluent to obtain the purified blank aquatic sample, which is the blank sample matrix. The blank aquatic sample was used in a ratio of 1g to 10ml of ethyl acetate. The methanol-acetonitrile aqueous solution was composed of methanol, acetonitrile, and water, with a volume ratio of 1:0.21 for methanol and 1:1 for acetonitrile and water. The blank aquatic sample was used in a ratio of 1g to 1ml of methanol-acetonitrile aqueous solution for redissolution. The blank aquatic sample was used in a ratio of 1g to 0.5ml of acidified methanol. The acidified methanol was composed of acetic acid and methanol, with a volume ratio of 1:99 for acetic acid and methanol. The blank aquatic sample was a black carp sample. The solid-phase extraction column was the solid-phase extraction column prepared in Examples 6-10 and Comparative Examples 1-2.

[0137] To investigate the matrix effect, 18 hormonal drug standards were prepared into mixed standard solutions using both blank sample matrix and methanol-acetonitrile aqueous solution as solvents. The concentrations of the 18 hormonal drug standards in the mixed standard solutions were 0, 1, 2, 5, 10, 20, 50, and 100 ng / mL. Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) was used for detection. The matrix effect was evaluated by the percentage slope (SR) of the standard curves for both the blank matrix solution and the methanol-acetonitrile aqueous solution. A SR closer to 100% indicated a smaller matrix effect; SR < 100% indicated a matrix inhibition effect; and SR > 100% indicated a matrix enhancement effect.

[0138] Table 4. SR value results for hydrocortisone-based hormone drugs.

[0139]

[0140] The SR values ​​for hydrocortisone hormone drugs are shown in Table 4. Compared with Example 7, Example 6 shows that the use of modified polystyrene can further improve the purification effect of the prepared activated solid-phase extraction column on aquatic samples and reduce the matrix effect. Compared with Example 8, Example 7 shows that increasing the amount of modified polystyrene within a certain range can improve the purification effect of the prepared activated solid-phase extraction column on aquatic samples and reduce the matrix effect. Compared with Example 9, Example 7 shows that using poly(1,4-cyclohexanedimethyl adipate) to prepare nanofibers based on modified polystyrene and using them for subsequent preparation of activated solid-phase extraction columns can improve the purification effect of the prepared activated solid-phase extraction column on aquatic samples and further reduce the matrix effect. Compared with Example 10, Example 9 shows that poly(1,4-cyclohexanedimethyl adipate) Increasing the amount of modified polystyrene used within a certain range can improve the purification effect of the prepared activated solid-phase extraction column on aquatic samples and reduce the matrix effect. Compared with Comparative Example 1, Example 7 shows that the amount of modified polystyrene used needs to be within a suitable range. Too little modified polystyrene will reduce the purification effect of the prepared activated solid-phase extraction column on aquatic samples and enhance the matrix effect. Compared with Comparative Example 2, Example 9 shows that modified polystyrene and poly(1,4-cyclohexanedimethyl adipate) need to be used together. Using poly(1,4-cyclohexanedimethyl adipate) alone has no significant effect on reducing the matrix effect.

[0141] Experimental Example 5:

[0142] To determine the linearity, limit of detection (LOD), and limit of quantitation (LOQ), 18 hormone drug standards were prepared into a mixed standard solution using acetonitrile-water solution. The concentrations of the 18 hormone drug standards in the mixed standard solution were a series of 0, 1, 2, 5, 10, 20, 50, and 100 ng / mL. Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) was used for detection, and external standard method was used for quantification. A standard curve was plotted with the concentration of the 18 hormone drugs in the mixed standard solution as the abscissa (X, μg / L) and the corresponding peak area (Y) as the ordinate. Using the standard spiking method, the mixed standard solution was added to blank milk samples with progressively decreasing spiking amounts, and then detection was performed. The LOD and LOQ were calculated.

[0143] Table 5. Results of linearity, detection limit, and quantitation limit determination.

[0144]

[0145] The detection and calculation results are shown in Table 5. The 18 hormone drugs showed good linearity in the range of 1-100 μg / L, with a correlation coefficient (R0). 2All values ​​were greater than 0.99. The limits of detection for the 18 hormone drugs were 0.15-2.1 μg / kg, and the limits of quantitation were 0.3-4.2 μg / kg.

[0146] Experimental Example 6:

[0147] The recoveries and relative standard deviations were determined by preparing a mixed standard solution of 18 hormone drugs using acetonitrile-water solution. This mixed standard solution was then added to a blank aquatic sample to achieve a hormone drug concentration of 5 μg / kg, resulting in a spiked sample. Next, the purified aquatic sample to be tested was prepared. The preparation of the purified aquatic sample to be tested was the same as the preparation of the blank sample matrix in Example 4, except that the blank aquatic sample was replaced with the spiked sample, and the solid-phase extraction column was the one prepared in Example 6. The recoveries and relative standard deviations after spiked preparation of the 18 hormone drugs were calculated based on the standard curve.

[0148] Table 6. Results of recovery rate and relative standard deviation after adding 18 kinds of hormone drugs.

[0149]

[0150] The results are shown in Table 6. The recoveries of the 18 hormone drugs ranged from 76.7% to 94.6%, and the relative deviations ranged from 1.9% to 10.1%, indicating that the method has high accuracy and can effectively detect hormone drugs in aquatic products.

[0151] The embodiments and / or implementation methods described above are merely preferred embodiments and / or implementation methods for implementing the technology of the present invention, and are not intended to limit the implementation methods of the technology of the present invention in any way. Any person skilled in the art can make some modifications or alterations to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as the technology or embodiments that are substantially the same as the present invention.

[0152] This document uses specific examples to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the methods and core ideas of this application. The above descriptions are only preferred embodiments of this application. It should be noted that due to the limitations of written expression, while there are objectively infinite specific structures, those skilled in the art can make several improvements, modifications, or changes without departing from the principles of this application, and can also combine the above technical features in an appropriate manner. These improvements, modifications, changes, or combinations, or the direct application of the inventive concept and technical solution to other situations without modification, should all be considered within the scope of protection of this application.

Claims

1. A method for preparing an activated solid-phase extraction column, comprising: S1: The solvent is mixed to obtain a mixture, then the ligand and metal source are added and stirred, then the alkali is added and ultrasonic treatment is performed, and finally the 2D-MOF is obtained after washing. S2: 2D-MOF, polymer and solvent are mixed to obtain electrospinning solution. The electrospinning solution is electrospinned to obtain electrospinning film, which is dried to obtain nanofibers; the mass ratio of 2D-MOF to polymer is 1:1-5. S3: First, the tablet is fed to the lower end of the solid phase extraction column, then the nanofibers are placed in, and then the tablet is pressed to obtain the solid phase extraction column. The nanofibers are activated by flowing an activator through the column to obtain the activated solid phase extraction column. The polymer in S2 is modified polystyrene, which is prepared from 4-chlorobutyryl chloride, polystyrene, and 2,2'-(propane-2,2-dimethylbis(sulfonamide dimethyl))diethylamine; The preparation of modified polystyrene is as follows: aluminum chloride, 4-chlorobutyryl chloride and chloroform are mixed; then added dropwise to a polystyrene solution and reacted at 25-35℃ for 2-5 hours; after the reaction is completed, the mixture is washed, and then ethanol is added to separate the precipitate and dry it to obtain an intermediate; N,N-dimethylformamide and 2,2'-(propane-2,2-dimethylbis(sulfonamide dimethyl))diethylamine are added to the intermediate and reacted at 50-60℃ for 2-5 hours. Finally, the mixture is washed and dried to obtain modified polystyrene.

2. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, The solvent in S1 is N,N-dimethylformamide, ethanol, and water, with a volume ratio of N,N-dimethylformamide to ethanol of 1:0.03-0.1 and a volume ratio of N,N-dimethylformamide to water of 1:0.03-0.1; the ligand is terephthalic acid, with a volume ratio of terephthalic acid to N,N-dimethylformamide of 1 mmol:40-50 ml.

3. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, The metal source in S1 is at least one of MnCl2·4H2O, CoCl2·6H2O and NiCl2·6H2O, and the molar ratio of the amount of ligand to metal source is 1:0.5-1.

5.

4. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, The base in S1 is triethylamine, and the ratio of ligand to base is 1 mmol: 0.8-1.5 ml.

5. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, In S2, the polymer is modified polystyrene and poly(1,4-cyclohexanedimethyl adipate) in a mass ratio of 1:0.5-1.

2.

6. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, The solvent in S2 is N,N-dimethylformamide; the ratio of polymer to solvent is 1g:10-20ml.

7. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, The activator in S3 is methanol and water. Methanol and water flow sequentially through the nanofibers inside the column. The ratio of nanofibers to methanol is 1g:20-30ml, and the ratio of nanofibers to water is 1g:40-60ml.

8. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, The stirring time in S1 is 20-40 min, and the ultrasonic treatment time is 6-10 h.

9. The method for preparing an activated solid-phase extraction column according to claim 1, characterized in that, In S2, the curing distance of electrospinning is 15-25cm, the voltage is 15-25kv, and the injection speed is 40-50uL / min.

10. The application of the activated solid-phase extraction column prepared by any of the preparation methods of claims 1-9 in the preparation of purified aquatic samples.

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