CN115785646A

CN115785646A - Adsorption filler for purifying column and preparation method and application thereof

Info

Publication number: CN115785646A
Application number: CN202211491960.3A
Authority: CN
Inventors: 徐婷; 薛昆鹏; 李良翔; 任兴发; 屠炳芳; 陈超; 陈庆涛
Original assignee: Jiangsu Yuexu New Material Technology Co ltd; Welch Materials (shanghai) Inc; Zhejiang Yuexu Material Technology Co ltd
Current assignee: Jiangsu Yuexu New Material Technology Co ltd; Welch Materials (shanghai) Inc; Zhejiang Yuexu Material Technology Co ltd
Priority date: 2022-11-25
Filing date: 2022-11-25
Publication date: 2023-03-14
Also published as: CN117753373A

Abstract

The invention belongs to the field of chromatographic packing, and discloses an adsorption packing for a purification column, and a preparation method and application thereof. The composite material of polyethylene oxide and clay comprises the following raw materials in parts by weight: 4000-8000 portions of inorganic salt modified clay, 50-100 portions of quaternary ammonium salt compound and 1 portion of polyethylene oxide. The invention also provides an adsorption filler for the purification column, which can effectively and selectively adsorb impurities such as protein, lipid and the like in the extracting solution before mycotoxin detection, has smaller matrix effect and high absolute recovery rate on several typical mycotoxin targets.

Description

Adsorption filler for purification column and preparation method and application thereof

Technical Field

The invention relates to the field of chromatographic packing, in particular to an adsorption packing for a purification column and a preparation method and application thereof.

Background

The term Mycotoxin (Mycotoxin) is derived from the Greek word "Mykes" and the Latin word "Toxicum" and is a secondary metabolite produced by a toxigenic fungus under appropriate environmental conditions. Rice, wheat and corn are three kinds of cereal food which are most consumed in China, the cereal food is very easy to be polluted by mycotoxin in the processing or storage process, and the problem of fungal pollution is more and more widely concerned.

The aflatoxin is a secondary metabolite produced by aspergillus flavus, aspergillus parasiticus and the like in a high-temperature and humid environment, has toxicity, and is a highly toxic substance. Besides aflatoxin, more intensive researches are carried out on more than ten fungal toxins which are harmful to human beings, and the fungal toxins have the characteristics of strong toxicity and high pollution frequency, wherein deoxynivalenol (vomitoxin), zearalenone and patulin are the most prominent ones. The four mycotoxins not only have the effects of carcinogenesis, teratogenesis, mutagenesis and the like, but also have the effects of hepatotoxicity, toxic kidney damage, reproductive disorder, immunosuppression and the like, and pose great threat to human health. Mycotoxin infection in food has become a considerable problem, so that a rapid, efficient and sensitive detection method is established, and particularly, the simultaneous detection of multiple mycotoxins is very important.

At present, methods for detecting mycotoxins in food and agricultural products mainly comprise a thin-layer chromatography method, an enzyme-linked immunosorbent assay method, an immunoaffinity column purification-fluorescence method, an immunoaffinity column purification-high performance liquid chromatography method, a high performance liquid chromatography-tandem mass spectrometry method and the like. In order to reduce detection interference, improve detection capability of the method and prolong the service life of the instrument, the sample extracting solution needs to be purified before entering the instrument for analysis, namely, non-target substances such as fat, protein, pigment, mineral substances, cellulose and the like which are greatly existed in the sample extracting solution are removed in a certain mode. The current common purification means mainly comprise the following steps:

(1) Liquid-Liquid extraction (LLE) is a Liquid-Liquid extraction method which is based on the difference in solubility between impurities and a substance to be detected in different solvents to complete extraction and purification, and for example, a weak-polar solvent such as n-hexane, petroleum ether and the like can be used to remove polar impurities such as grease, cholesterol and the like in an extracting solution, while a polar target substance is retained. The liquid-liquid extraction method has the advantages of simplicity, low requirement on equipment, high operation speed and wide application, but has the defects of long time consumption, low efficiency, poor method pertinence, large solvent consumption and the like.

(2) Solid phase extraction (Solidphase extraction, SPE)

SPE is very similar to column chromatography in terms of separation principle, stationary phase and solvent selection, but SPE packing particle size (above 40 gm) is much larger than chromatography column packing, so that only substances with large differences in properties can be separated. The common packing material for SPE column is silica gel or chemical bond and packing material, such as-OH, -CN, -C18, -C ₆ H ₅ And the like, and the adsorption of the target substance is accomplished by a bond and a functional group. This can be done by SPE: 1) Removing matrix interferents; 2) Enriching trace components; 3) Changing the sample solvent; 4) In situ derivatization, sample desalinization, and the like. The general operational procedure of an SPE can be described as: 1) Activation ofRinsing the SPE column with a suitable solvent to remove impurities that may be present in the column and solvate the surface of the packing; 2) Loading the sample, and slowly passing the extracting solution through the SPE column under the action of negative pressure or gravity to achieve a process of full contact and distribution balance; 3) Leaching, eluting the interference substances remained on the column by using a medium-strength solvent, and continuously keeping the substances to be detected; 4) Elution, the test substance is eluted completely and collected in as small a fraction as possible.

Currently, the SPE method has been used in place of the original LLE method in many analytical methods and even in national standards. Compared with the LLE method, SPE has the following advantages: the dosage of the organic reagent is less, and the operation process is quick; the method has good selectivity and high sensitivity; the repeatability is good, and the automation is easy to realize; can process large-volume samples and has the function of concentration and enrichment. However, the SPE method also has some disadvantages: the column itself is easily blocked and the filler selection range is limited; column performance is affected by factors such as pH, solvent type, and ionic strength.

(3) Improved solid phase extraction process

The improved solid phase extraction method is similar to the traditional SPE method in principle, but has difference on the types of fillers and purification mechanism, and mainly comprises two methods: immunoaffinity columns (IAC) and Multifunctional purification columns (MFC).

The multifunctional purifying column is a more popular purifying column in recent years, and the action principle of the multifunctional purifying column is mainly physical adsorption. Polar, nonpolar and ion exchangers are usually used as adsorbents to selectively adsorb impurities such as proteins and lipids in the extract when the extract contacts with the filler, while the toxin to be detected is not retained. Compared with the traditional SPE method, the MFC operation is quicker, the processes of activation, leaching, elution and the like are not needed, the sample flux is large, the MFC is suitable for simultaneous determination of various toxins, and compared with the IAC, the MFC has the advantages of lower detection cost and high experimental efficiency, and is suitable for determination of a large number of samples; but the disadvantages of MFC are also evident: the purification effect on complex matrix is limited, larger matrix effect still exists, and the absolute recovery rate of part of target substances is poor.

Disclosure of Invention

In view of the above-mentioned disadvantages of the prior art, the present invention aims to provide an adsorption packing for a purification column, a preparation method and a use thereof, so as to solve the problems in the prior art.

In order to achieve the purpose, the invention adopts the following technical scheme.

The first aspect of the invention protects a composite material of polyethylene oxide and clay, which comprises the following raw materials in parts by weight:

4000-8000 parts of inorganic salt modified clay

50-100 parts of quaternary ammonium salt compound

1 part of polyethylene oxide.

In some embodiments, the inorganic salt modified clay may be present in an amount of 4000 to 6000 parts by weight, or 5000 to 7000 parts by weight, or 6000 to 8000 parts by weight. In a preferred embodiment, 6000 parts.

In some embodiments, the weight part of the quaternary ammonium salt compound may be 50 to 110 parts, 80 to 150 parts, or 120 to 200 parts. In a preferred embodiment, the amount is 100 parts.

The composite material of the invention, inorganic salt modified clay, increases the expansibility of clay, embeds the cations of quaternary ammonium salt compound between crystal layers of inorganic salt modified clay to obtain organic clay, and initiates the polymerization of polyethylene oxide between the crystal layers, which penetrates into the clay layers, to successfully prepare the composite material of polyethylene oxide and clay. The composite material has uniform particle size, and can selectively adsorb protein and lipid in mycotoxin extracting solution.

In certain embodiments, the quaternary ammonium salt compound is selected from one or more of N, N-trimethyl-2- (2-methyl-1-oxo-2-propenyloxy) ethylammonium chloride-acrylamide copolymer, 2-hydroxy-3- (trimethylamino) propylpolyethylene oxide cellulose ether chloride, alkyl dimethyl benzyl ammonium halide, and dialkyl dimethyl ammonium halide.

In some embodiments, the polyethylene oxide has a weight average molecular weight to number average molecular weight ratio of 17 to 20, a particle size of 300 to 400 mesh, and a dissolution temperature of 30 to 50 ℃.

In certain embodiments, the inorganic salt modified clay is selected from one or more of a sodium modified clay, a potassium modified clay, and a calcification modified clay.

Preferably, the inorganic salt modified clay is a mixture of sodium modified clay, potassium modified clay and calcification modified clay. The cation modified clay composed of the sodium modified clay, the potassium modified clay and the calcification modification can improve the viscosity and the lubricity of the clay, is beneficial to preparing a filler, and has the advantages of difficult adhesion of a polymer and good selective adsorption.

More preferably, the mass ratio of the sodium modified clay to the potassium modified clay to the calcification modified clay is (0.5-2): (0.5-2) 1. More preferably, the mass ratio of the sodium modified clay, the potassium modified clay and the calcification modified clay may be (0.5 to 0.8): (0.5-2): 1, and may be (0.6-1.5): 1 in (0.5-2), or (1.2-2): (0.5-2) 1. In a preferred embodiment, the ratio of 1:1:1.

in some embodiments, the inorganic salt modified clay is prepared by:

1) Dispersing clay in water, performing ultrasonic treatment, and centrifuging to obtain a suspension;

2) Adjusting the pH value of the suspension by adopting acid, and adding an oxidant for reaction;

3) And adjusting the pH value by adopting alkali, adding inorganic salt, mixing, centrifuging and drying to obtain the inorganic salt modified clay.

According to the invention, firstly, the clay is pretreated, and impurity mineral particles are removed through ultrasound and centrifugation, so that a foundation is laid for subsequently increasing the contact area between the clay and a polymer, and meanwhile, the interlamellar spacing of clay crystals is increased, so that a foundation is laid for the subsequent polymer to enter the interlamellar space of the clay crystals; the specific surface area of the clay is increased through the synergistic treatment of acidification and oxidation, organic matters contained in the clay are removed, holes are enlarged due to the removal of the organic matters, and an environment is created for subsequent hydration and expansion; and adding sodium carbonate later to ensure that the solution is changed into an alkaline environment, and then adding salt, wherein the step is to better insert cations and increase the expansibility of the clay, which comprises the inner part of the clay and the outer part of the clay, so that conditions are created for the subsequent polymer to enter a clay crystal layer, and the subsequent quaternary ammonium salt compound is convenient to enter.

Preferably, in 1), the clay is selected from one or more of kaolinite, montmorillonite, serpentine, talc and mica.

Preferably, in 2), the pH value is 2 to 7. More preferably, the pH may be 2 to 4, 3 to 6, or 5 to 7. In a preferred embodiment, the number is 6.

Preferably, in 2), the acid is selected from one or more of dilute hydrochloric acid, dilute nitric acid and dilute sulfuric acid.

More preferably, the acid is dilute hydrochloric acid. Further preferably, the concentration of the dilute hydrochloric acid is 1 to 10mol/L. Preferably, the concentration of the dilute hydrochloric acid can be 1 to 5mol/L, also can be 4 to 8mol/L, also can be 6 to 10mol/L. In a preferred embodiment, the concentration is 5mol/L.

Preferably, in 2), the oxidant is selected from one or more of hydrogen peroxide, peracetic acid and nitric acid.

More preferably, the oxidizing agent is hydrogen peroxide (H) ₂ O ₂ )。

Preferably, in 3), the pH value is 8-10. More preferably, the pH may be 8 to 8.8, 8.6 to 9.6, or 9.2 to 10. In a preferred embodiment, 10.

Preferably, in 3), the alkali is selected from one or two of sodium carbonate and sodium bicarbonate.

More preferably, the base is sodium carbonate.

Preferably, in 3), the inorganic salt is selected from one or more of sodium chloride, potassium chloride and calcium chloride.

Preferably, the mass ratio of the clay to the inorganic salt is (5-7): (12 to 30).

More preferably, when the inorganic salt is sodium chloride, the mass ratio of the clay to the sodium chloride is (5-7): (12 to 25).

More preferably, when the inorganic salt is potassium chloride, the mass ratio of the clay to the potassium chloride is (5-7): (18 to 30).

More preferably, when the inorganic salt is calcium chloride, the mass ratio of the clay to the calcium chloride is (5-7): (19 to 25).

Preferably, the mass ratio of the clay to the oxidant is 1: (2 to 20).

More preferably, the mass ratio of the clay to the oxidizing agent may be 1: (2-10), may be 1: (6 to 15), which may be 1: (12 to 20). In a preferred embodiment, the ratio is 1:12.

a second aspect of the present invention provides a method for preparing the above composite material, comprising: inorganic salt modified clay and quaternary ammonium salt compound react in solvent and then react with polyethylene oxide to obtain the composite material.

The inorganic salt modified clay is placed in the aqueous solution of the quaternary ammonium salt compound, so that the inorganic salt clay is modified from polarity to non-polarity, conditions are created for the subsequent coating of polyethylene oxide, and the adsorption of the composite material of polyethylene oxide and clay to oil, protein and lipid is increased.

In certain embodiments, the solvent is water.

In certain embodiments, the reaction temperature is 40 to 80 ℃.

Preferably, the reaction temperature can be 40-55 ℃, also can be 50-75 ℃, also can be 60-80 ℃. In a preferred embodiment, it is 60 ℃.

In some embodiments, the reaction time is 1 to 10 hours.

Preferably, the reaction time may be 1 to 6 hours, 4 to 8 hours, or 6 to 10 hours. In a preferred embodiment, the time is 5 hours.

In certain embodiments, the polymerization temperature is from 20 to 40 ℃.

Preferably, the polymerization reaction temperature can be 20-28 ℃, 26-36 ℃ or 35-40 ℃. In a preferred embodiment, 25 ℃.

In some embodiments, the polymerization reaction time is from 5 to 8 hours.

Preferably, the reaction time may be 5 to 6.2 hours, 5.8 to 7.2 hours, or 7.0 to 8 hours. In a preferred embodiment, the time period is 7 hours.

A third aspect of the invention protects the use of the composite material described above as a raw material in an adsorbent filler.

The fourth aspect of the present invention protects an adsorbent packing for a purification column, which comprises the above-described composite material, activated silica gel, and a silane coupling agent.

In some embodiments, the adsorbent packing for purification column comprises the following raw materials in parts by weight:

40 to 70 portions of composite material

20 to 50 portions of activated silica gel

5-12 parts of a silane coupling agent.

In the application, the silica gel and the polymer are compounded to adsorb partial grease, but can also adsorb a target object, and after the composite material is added, interferents such as grease and the like can be adsorbed to the maximum extent, and the target object is not adsorbed.

Preferably, the weight part of the composite material can be 40 to 58 parts, 48 to 62 parts or 55 to 70 parts. In a preferred embodiment, the amount is 40 parts, 60 parts or 70 parts.

Preferably, the activated silica gel may be 20 to 24 parts by weight, 23 to 26 parts by weight, or 25 to 30 parts by weight. In a preferred embodiment, the amount is 50 parts, 30 parts, 20 parts or 25 parts.

Preferably, the silane coupling agent may be present in an amount of 5 to 7 parts by weight, 6 to 10 parts by weight, or 9 to 12 parts by weight. In a preferred embodiment, the amount is 10 parts or 5 parts.

In certain embodiments, the silane coupling agent is selected from one or more of gamma-aminopropyltriethoxysilane (KH-550), vinyltriethoxysilane (A151), N- (. Beta. -aminoethyl) -gamma-aminopropyltrimethoxysilane (KH-792), gamma-aminoethylaminopropyltrimethoxysilane (KH-900).

In certain embodiments, the activated silica gel is prepared by a method comprising: mixing the silica gel raw powder with acid in a solvent, and drying to obtain the activated silica gel.

In some embodiments, the silica gel raw powder has a specific surface area of 300 to 680m ² /g。

In some embodiments, the silica gel raw powder has a pore size of 10 to 25nm.

In certain embodiments, the acid is selected from one or more of sulfuric acid, hydrochloric acid, nitric acid, and perchloric acid.

Preferably, the acid is sulfuric acid.

In certain embodiments, the solvent is water.

In some embodiments, the mass volume ratio of the silica gel raw powder to the acid is (1-8) g:1mL.

Preferably, the mass volume ratio of the silica gel raw powder to the acid can be (1-5) g:1mL, and may be (3 to 7) g:1mL, or (6-8) g:1mL. In a preferred embodiment, the ratio is 4g:1mL.

In certain embodiments, the mixing temperature is 40 to 80 ℃.

Preferably, the mixing temperature can be 40-60 ℃, also can be 50-70 ℃, and also can be 60-80 ℃. In a preferred embodiment, 60 ℃.

In some embodiments, the mixing time is 2 to 8 hours.

Preferably, the mixing time may be 2 to 5 hours, 4 to 6 hours, or 5 to 8 hours. In a preferred embodiment, it is 4h.

In some embodiments, the drying temperature is 5 to 20 hours.

Preferably, the drying time may be 5 to 10 hours, 8 to 15 hours, or 12 to 20 hours. In a preferred embodiment, it is 10 hours.

A fifth aspect of the present invention protects the method for preparing an adsorption packing for a purification column as described above, comprising the steps of: and mixing the raw material components, and aging to obtain the adsorption filler for the purification column.

In certain embodiments, the aging is performed in an aqueous sodium hydroxide solution.

Preferably, the concentration of the sodium hydroxide aqueous solution is 0.1-1.0 mol/L. More preferably, the concentration may be 0.1 to 0.4mol/L, 0.3 to 0.7mol/L, or 0.6 to 1.0mol/L. In a preferred embodiment, the concentration is 0.8mol/L.

In some embodiments, the aging time is from 4 to 10 hours.

Preferably, the aging time may be 4 to 10 hours, or 4 to 10 hours. In a preferred embodiment, the time is 5 hours.

A sixth aspect of the invention protects the use of an adsorbent packing for a purification column as described above as a chromatographic material.

In certain embodiments, the use of the adsorbent packing for a purification column as an adsorbent in the preparation of a chromatography material for mycotoxins.

The adsorption filler for the purification column can effectively and selectively adsorb impurities such as protein, lipid and the like in the mycotoxin pretreatment extracting solution, so that the chromatographic analysis of trace or trace substances is more accurate.

Compared with the prior art, the invention has the following beneficial effects:

1) The purifying column of the present invention compounds the composite material of polyethylene oxide and clay and activated silica gel with the adsorbing filler through silane coupling agent, and can perform effective purification pretreatment to mycotoxin.

2) The adsorption filler for the purification column is very suitable for the requirement of mycotoxin detection, and can effectively and selectively adsorb impurities such as protein, lipid and the like in the pretreatment extracting solution, so that the chromatographic analysis of trace or trace substances is more accurate, and the determination of several mycotoxins is realizedThe amount of the component to be detected in the mixture is 10 ^-6 ～10 ^-9 (mass ratio).

3) The adsorption filler for the purification column has good purification effect on complex matrixes, has smaller matrix effect and has high absolute recovery rate on several typical mycotoxin target objects.

4) The adsorption filler for the purification column shows uniformity and stability in particle size distribution and pore size distribution.

Drawings

FIG. 1 is a graph showing the recovery of the test toxin from the spiked extract by the adsorption packing for purification columns of examples 1 to 10 of the present invention.

FIG. 2 shows that the adsorbing filler for purification column of example 10 of the present invention, the fillers of comparative example 1 and comparative example 2 are against aflatoxin B, respectively ₁ Aflatoxins B ₂ Aflatoxin G ₁ And aflatoxin G ₂ The solvent effect evaluation chart was performed.

Fig. 3 is a microscopic view showing the adsorbing filler for purification column of example 10 of the present invention, and the fillers of comparative example 1 and comparative example 2.

Fig. 4 is a schematic view showing the batch stability effect of the adsorption packing for purification columns according to example 10 of the present invention.

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not to be limited to the specific embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the respective manufacturers.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

In the following examples and comparative examples of the present application, the sources and parameters of the respective raw material components used are shown in Table 1.

TABLE 1 Source of raw materials and parameters

Example 1

In this embodiment, a composite material of polyethylene oxide and clay, an adsorption filler for a purification column, and a preparation method thereof are provided, including the following steps.

The formula of the composite material of the polyethylene oxide and the clay is as follows: 60g of inorganic salt-modified clay, 2g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether and 10mg of polyethylene oxide, wherein the inorganic salt-modified clay comprises 20g of sodium-modified clay, 20g of potassium-modified clay and 20g of calcification-modified clay.

A method of making a composite of polyethylene oxide and clay comprising the steps of:

1) 700g of kaolinite are dispersed in 2L of deionized water, stirred at 7000rpm for 30min, sonicated for 15min, and then centrifuged at 5000rpm for 80min to remove precipitates and remove other mineral particles, resulting in a suspension.

2) Adjusting the pH of the suspension to 2-7 with 5mol/L dilute hydrochloric acid, adding 10mL of 10% ₂ O ₂ Heating to 80 ℃ for H ₂ O ₂ Decomposing to remove organic matter in kaolinite, and regulating pH value to 9-10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added separately ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, taking precipitate, washing the precipitate until no redundant solution exists, drying and crushing at 120 ℃, and sieving with a 500-mesh sieve to respectively prepare the sodium modified kaolinite, the potassium modified kaolinite and the calcified modified kaolinite.

4) Respectively placing 20g of sodium modified kaolinite, 20g of potassium modified kaolinite and 2g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether in 500mL of distilled water, carrying out water bath reaction at 60 ℃ for 5h, cooling to room temperature (25 ℃) after the reaction is finished, adding 10mg of polyethylene oxide for carrying out polymerization reaction for 7h, and carrying out ultrasonic stirring to obtain reaction products; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (called composite kaolin polymer for short).

The formula of the adsorption filler for the purification column comprises: 40wt% of a composite of polyethylene oxide and clay (referred to as composite kaolin polymer for short), 50wt% of activated silica gel and 10wt% of KH-550.

The preparation method of the adsorption filler for the purification column comprises the following steps:

stirring the composite kaolin polymer, the activated silica gel and the silane coupling agent (KH-550) in a reaction kettle for 2 hours by using a magnetic stirrer, wherein the stirring speed is 100r/min, so as to obtain a mixed solution; aging 600mL of the mixed solution in 1000mL of 0.5mol/L sodium hydroxide solution for 5h to obtain microspheres, separating the microspheres from the aged solution by using a screen, washing the microspheres with a washing solution such as deionized water for 5 times, and drying the microspheres at 50-65 ℃ to obtain the adsorption filler for the purification column, wherein the adsorption filler is marked as adsorption filler No. 1.

The activated silica gel is obtained by the following method:

taking the specific surface area of 300-680 m ² 200g of silica gel with the pore diameter distribution of 10-25 nm, mixing with 50mL of concentrated sulfuric acid and 200mL of distilled water in a 1000mL two-neck flask, magnetically stirring and soaking in a constant-temperature water bath at 60 ℃ for 4 hours, washing with distilled water, filtering to neutrality, drying in an oven at 75 ℃ for 10 hours, and cooling for later use.

Example 2

In this embodiment, a composite material of polyethylene oxide and clay, an adsorption filler for a purification column, and a preparation method thereof are provided, which include the following steps.

The formula of the composite material of the polyethylene oxide and the clay is as follows: 60g of inorganic salt modified clay, 1g of dialkyl dimethyl ammonium halide and 10mg of polyethylene oxide, wherein the inorganic salt modified clay comprises 20g of sodium modified clay, 20g of potassium modified clay and 20g of calcification modified clay.

1) 700g of montmorillonite was dispersed in 2L of deionized water, stirred at 7000rpm for 30min, sonicated for 15min, and then centrifuged at 5000rpm for 80min to remove precipitates and remove other mineral particles, resulting in a suspension.

2) The pH of the suspension is initially adjusted to 5 with 5mol/L dilute hydrochloric acid, 15ml of 10% are added ₂ O ₂ Heating to 80 ℃ for H ₂ O ₂ Decomposing to remove organic substances in montmorillonite, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added separately ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution is left, oven drying and pulverizing at 120 deg.C, and sieving with 500 mesh sieve to obtain sodium modified montmorillonite, potassium modified montmorillonite and calcified modified montmorillonite respectively.

4) Respectively 20g of sodium modified montmorillonite, potassium modified montmorillonite and calcified modified montmorillonite and 1g of dialkyl dimethyl ammonium halide are placed in 500mL of distilled water to react for 5 hours in a water bath at 60 ℃, 10mg of polyethylene oxide is added after the reaction is finished and cooled to room temperature (25 ℃), and the polymerization reaction is carried out for 7 hours by ultrasonic stirring to obtain a reaction product; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression and drying, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (called composite montmorillonite polymer for short).

The formula of the adsorption filler for the purification column comprises: 40wt% of a composite of polyethylene oxide and clay (abbreviated as composite montmorillonite polymer), 50wt% of activated silica gel and 10wt% of A151.

stirring the composite montmorillonite polymer, the activated silica gel and the silane coupling agent (A151) in a reaction kettle for 2 hours by a magnetic stirrer at the stirring speed of 100r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 5h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorption filler for the purification column, wherein the adsorption filler is marked as adsorption filler # 2.

The activated silica gel was prepared in the same manner as in example 1.

Example 3

The formula of the composite material of polyethylene oxide and clay is as follows: 60g of inorganic salt modified clay, 1g of alkyldimethylbenzyl ammonium halide and 10mg of polyethylene oxide, wherein the inorganic salt modified clay comprises 20g of sodium modified clay, 20g of potassium modified clay and 20g of calcification modified clay.

1) 700g of serpentine is dispersed in 2L of deionized water, stirred at 10000rpm for 50min, sonicated for 15min, and then centrifuged at 8000rpm for 40min to remove precipitates and remove other mineral particles, thus obtaining a suspension.

2) The suspension is initially adjusted to pH 5 with 5mol/L dilute hydrochloric acid and 15mL of 10% H are added ₂ O ₂ Heating to 90 ℃ for H ₂ O ₂ Decomposing to remove organic substances in serpentine, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added respectively ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution is left, oven drying at 120 deg.C, pulverizing, sieving with 500 mesh sieve, and respectively making into sodium modified snakeSerpentine, sylvite modified serpentine, and calcified modified serpentine.

4) Respectively placing 20g of sodium modified serpentine, potassium modified serpentine and calcified modified serpentine and 1g of alkyl dimethyl benzyl ammonium halide in 500mL of distilled water, carrying out water bath reaction at 60 ℃ for 5h, cooling to room temperature (25 ℃) after the reaction is finished, adding 10mg of polyethylene oxide, and carrying out ultrasonic stirring for polymerization reaction for 7h to obtain a reaction product; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (called composite serpentine polymer for short).

The formula of the adsorption filler for the purification column comprises: 60wt% composite of polyethylene oxide and clay (composite serpentine polymer), 30wt% activated silica gel and 10wt% A151.

stirring the composite serpentine polymer, the activated silica gel and the silane coupling agent (A151) in a reaction kettle for 2 hours by a magnetic stirrer at the stirring speed of 180r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorbent filler for purification column labeled as adsorbent filler # 3.

The activated silica gel was prepared as in example 1.

Example 4

The formula of the composite material of polyethylene oxide and clay is as follows: 60g of inorganic salt-modified clay, 1g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether and 10mg of polyethylene oxide, wherein the inorganic salt-modified clay comprises 20g of sodium-modified clay, 20g of potassium-modified clay and 20g of calcification-modified clay.

A method of making a composite of polyethylene oxide and clay comprising the steps of: 1) 700g talc was dispersed in 2L deionized water, stirred at 10000rpm for 50min, sonicated for 15min, and then centrifuged at 8000rpm for 40min to remove precipitates to remove other mineral particles, resulting in a suspension.

2) The suspension is initially adjusted to pH 5 with 5mol/L dilute hydrochloric acid, 15mL of 10% ₂ O ₂ Heating to 90 ℃ for H ₂ O ₂ Decomposing to remove organic matter in the talc, and adjusting the pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added separately ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution exists, drying and crushing at 120 ℃, and sieving with a 500-mesh sieve to obtain the sodium modified talc, the potassium modified talc and the calcification modified talc respectively.

4) Respectively placing 20g of sodium modified talc, 20g of potassium modified talc and 1g of chloridized-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether in 500mL of distilled water, carrying out water bath reaction at 60 ℃ for 5h, cooling to room temperature after the reaction is finished, then adding 10mg of polyethylene oxide, and carrying out ultrasonic stirring for polymerization reaction for 7h to obtain a reaction product; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (composite talc polymer for short).

The formula of the adsorption filler for the purification column comprises: 60wt% of a composite of polyethylene oxide and clay (abbreviated as composite talc polymer), 30wt% of activated silica gel and 10wt% of A151.

stirring the composite talc polymer, the activated silica gel and the silane coupling agent (A151) in a reaction kettle for 2 hours by using a magnetic stirrer, wherein the stirring speed is 180r/min, so as to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorbent filler for purification column labeled as adsorbent filler No. 4.

The activated silica gel was prepared in the same manner as in example 1.

Example 5

In this embodiment, a composite material of polyethylene oxide and clay, an additional filler for a purification column, and a method for producing the same are provided.

The formula of the composite material of the polyethylene oxide and the clay is as follows: 60g of inorganic salt-modified clay, 1g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether and 10mg of polyethylene oxide, wherein the inorganic salt-modified clay comprises 20g of sodium-modified clay, 20g of potassium-modified clay and 20g of calcification-modified clay.

A method of making a composite of polyethylene oxide and clay comprising the steps of: 1) Dispersing 700g mica in 2L deionized water, stirring at 10000rpm for 50min, performing ultrasonic treatment for 15min, centrifuging at 8000rpm for 40min, and removing precipitate to remove other mineral particles to obtain suspension.

2) The suspension is initially adjusted to pH 5 with 5mol/L dilute hydrochloric acid, 15mL of 10% ₂ O ₂ Heating to 90 ℃ for H ₂ O ₂ Decomposing to remove organic matter in mica, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added, respectively ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution exists, drying and crushing at 120 ℃, and sieving with a 500-mesh sieve to respectively obtain sodium modified mica, potassium modified mica and calcification modified mica.

4) Respectively 20g of sodium modified mica, potassium modified mica and calcified modified mica and 1g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether are put into 500mL of distilled water to react for 5h in a water bath at 60 ℃, 10mg of polyethylene oxide is added after the reaction is finished and cooled to room temperature (25 ℃), and the mixture is ultrasonically stirred to carry out polymerization reaction for 7h, so that a reaction product is obtained; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (called composite mica polymer for short).

The formula of the adsorption filler for the purification column comprises: 60wt% of a composite material of polyethylene oxide and clay (for short, composite mica polymer for purification column), 30wt% of activated silica gel and 10wt% of A151.

stirring the composite mica polymer, the activated silica gel and the silane coupling agent (A151) in a reaction kettle for 2 hours by a magnetic stirrer at the stirring speed of 180r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorption filler for the purification column, wherein the adsorption filler is marked as adsorption filler # 5.

The activated silica gel was prepared in the same manner as in example 1.

Example 6

The formula of the composite material of polyethylene oxide and clay is as follows: 60g of inorganic salt-modified clay comprising 20g of sodium-modified clay, 20g of potassium-modified clay and 20g of calcified modified clay, 1g of chlorinated-2-hydroxy-3- (trimethylamino) propylpolyethylene oxide cellulose ether and 10mg of polyethylene oxide.

1) 700g of kaolinite was dispersed in 2L of deionized water, stirred at 9000rpm for 50min, sonicated for 15min, and then centrifuged at 6000rpm for 70min to remove precipitates and remove other mineral particles, resulting in a suspension.

2) Adjusting the pH of the suspension to 6 with 5mol/l (strength) dilute hydrochloric acid, adding 15mL of 10% ₂ O ₂ Heating to 95 ℃ for H ₂ O ₂ Decomposing to remove organic substances in kaolinite, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added separately ₂ Stirring at room temperature for 48 hr to perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution is present, and oven-drying at 120 deg.CCrushing, and sieving with a 500-mesh sieve to obtain sodium modified kaolinite, potassium modified kaolinite and calcified modified kaolinite respectively.

4) Respectively 20g of sodium modified kaolinite, 20g of potassium modified kaolinite and 1g of calcified modified kaolinite and 1g of chlorinated 2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether are put into 500mL of distilled water to react for 5 hours in a water bath at 60 ℃, after the reaction is finished, the mixture is cooled to room temperature, 10mg of polyethylene oxide is added, and the mixture is ultrasonically stirred to carry out polymerization reaction for 7 hours, so as to obtain a reaction product; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (called composite kaolinite polymer for short).

The formula of the adsorption filler for the purification column is as follows: 70wt% of a composite of polyethylene oxide and clay (referred to simply as a composite kaolinite polymer), 20wt% of activated silica gel and 10wt% of KH-792.

stirring the compound kaolinite polymer, the activated silica gel and the silane coupling agent (KH-792) in a reaction kettle for 5 hours by using a magnetic stirrer at the stirring speed of 150r/min to obtain a mixed solution; aging the mixed solution in 0.8mol/L sodium hydroxide solution for 5h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorption filler for the purification column, wherein the adsorption filler is labeled as adsorption filler No. 6.

The activated silica gel was prepared in the same manner as in example 1.

Example 7

1) 700g of mica was dispersed in 2L of deionized water, stirred at 9000rpm for 50min, sonicated for 15min, and then centrifuged at 6000rpm for 70min to remove precipitates and remove other mineral particles, resulting in a suspension.

2) Adjusting the pH of the suspension to 5 with 5mol/L (concentration) dilute hydrochloric acid, adding 15mL of 10% ₂ O ₂ Heating to 95 ℃ to cause H ₂ O ₂ Decomposing to remove organic matter in mica, and adjusting pH to 10 with sodium carbonate.

4) Respectively 20g of sodium modified mica, 20g of potassium modified mica and 20g of calcified modified mica and 1g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether are put into 500mL of distilled water to be subjected to water bath reaction at 60 ℃ for 3h, after the reaction is finished, the mixture is cooled to room temperature (25 ℃), 10mg of polyethylene oxide is added, and the mixture is subjected to ultrasonic stirring to perform polymerization reaction for 7h to obtain a reaction product; and centrifugally separating a reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the product by using a 300-mesh sieve to obtain the composite material (the composite mica polymer for short) of the polyethylene oxide and the clay.

The formula of the adsorption filler for the purification column comprises: 70wt% of a composite material of polyethylene oxide and clay (abbreviated as composite mica polymer), 25wt% of activated silica gel and 5wt% of KH-792.

stirring the composite mica polymer, the activated silica gel and the silane coupling agent (KH-792) in a reaction kettle for 6 hours by using a magnetic stirrer at the stirring speed of 150r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorption filler for the purification column, wherein the adsorption filler is marked as adsorption filler No. 7.

The activated silica gel was prepared in the same manner as in example 1.

Example 8

1) 700g of serpentine was dispersed in 2L of deionized water, stirred at 9000rpm for 70min, sonicated for 15min, and then centrifuged at 6000rpm for 70min to remove precipitates and remove other mineral particles, resulting in a suspension.

2) Adjusting the pH of the suspension to 7 with 5mol/L dilute hydrochloric acid, adding 15mL of 10% ₂ O ₂ Heating to 95 ℃ to cause H ₂ O ₂ Decomposing to remove organic substances in serpentine, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added respectively ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution exists, drying and crushing at 120 ℃, and sieving with a 500-mesh sieve to obtain sodium modified serpentine, potassium modified serpentine and calcified modified serpentine respectively.

4) Respectively placing 20g of sodium modified serpentine, potassium modified serpentine and calcified modified serpentine and 1g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether in 500mL of distilled water, carrying out water bath reaction for 3h at 60 ℃, cooling to room temperature (25 ℃) after the reaction is finished, adding 10mg of polyethylene oxide, and carrying out ultrasonic stirring for polymerization reaction for 7h to obtain a reaction product; and centrifugally separating a reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the product by using a 300-mesh sieve to obtain the composite material of the polyethylene oxide and the clay (the composite serpentine polymer for short).

The formula of the adsorption filler for the purification column is as follows: 70wt% of complex serpentine polymer, 25wt% of activated silica gel and 5wt% of KH-792.

stirring the composite serpentine polymer, the activated silica gel and the silane coupling agent (KH-792) for 6 hours in a reaction kettle by using a magnetic stirrer at the stirring speed of 150r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorbent filler labeled as adsorbent filler No. 8 for the purification column.

The activated silica gel was prepared in the same manner as in example 1.

Example 9

1) 700g talc was dispersed in 2L deionized water, stirred at 9000rpm for 70min, sonicated for 15min, and then centrifuged at 6000rpm for 70min to remove precipitates and remove other mineral particles to give a suspension.

2) Adjusting the pH of the suspension to 7 with 5mol/L dilute hydrochloric acid, adding 15mL of 10% ₂ O ₂ Heating to 95 ℃ to cause H ₂ O ₂ Decomposing to remove organic substances in the talc, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added, respectively ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution is left, drying and crushing at 120 ℃, and sieving with a 500-mesh sieve to obtain modified talc, potassium modified talc and calcium modified talc respectively.

4) Respectively placing 20g of sodium modified talc, 20g of potassium modified talc and 1g of chloridized-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether in 500mL of distilled water, carrying out water bath reaction at 60 ℃ for 3h, cooling to room temperature (25 ℃) after the reaction is finished, adding 10mg of polyethylene oxide, and carrying out ultrasonic stirring to carry out polymerization reaction for 7h to obtain a reaction product; and centrifugally separating the reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the dried reaction product by using a 300-mesh sieve to obtain the composite material of polyethylene oxide and clay (the composite talc polymer for short).

The formula of the adsorption filler for the purification column is as follows: 70wt% of composite talc polymer, 25wt% of activated silica gel and 5wt% of KH-900.

stirring the composite talcum polymer, the activated silica gel and the silane coupling agent (KH-900) in a reaction kettle for 6 hours by a magnetic stirrer at the stirring speed of 150r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorption filler for the purification column, wherein the adsorption filler is marked as adsorption filler # 9.

The activated silica gel was prepared in the same manner as in example 1.

Example 10

The formula of the composite material of the polyethylene oxide and the clay is as follows: 60g of inorganic salt-modified clay comprising 20g of sodium-modified clay, 20g of potassium-modified clay and 20g of calcified modified clay, 1g of chlorinated-2-hydroxy-3- (trimethylamino) propylpolyethylene oxide cellulose ether and 10mg of polyethylene oxide.

1) 700g of montmorillonite was dispersed in 2L of deionized water, stirred at 9000rpm for 70min, sonicated for 15min, and then centrifuged at 6000rpm for 70min to remove precipitates and remove other mineral particles, resulting in a suspension.

2) Adjusting the pH of the suspension to 7 with 5mol/L dilute hydrochloric acid, adding 15mL of 10% ₂ O ₂ Reacting at 95 ℃ to cause H ₂ O ₂ Decomposing to remove organic substances in montmorillonite, and adjusting pH to 10 with sodium carbonate.

3) 300mL of 3M NaCl, 300mL of 3M KC1 and 300mL of 0.5M CaCl were added respectively ₂ Stirring at room temperature for 48h to fully perform ion exchange, centrifuging, collecting precipitate, washing the precipitate until no excess solution is left, drying and crushing at 120 ℃, and sieving with a 500-mesh sieve to obtain sodium modified montmorillonite, potassium modified montmorillonite and calcified modified montmorillonite respectively.

4) Respectively placing 20g of sodium modified montmorillonite, 20g of potassium modified montmorillonite and 20g of calcified modified montmorillonite and 1g of chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether into 500mL of distilled water, carrying out water bath reaction at 60 ℃ for 3h, cooling to room temperature (25 ℃) after the reaction is finished, adding 10mg of polyethylene oxide, and carrying out ultrasonic stirring for polymerization reaction for 7h to obtain a reaction product; and centrifugally separating a reaction product, washing the reaction product for many times by using distilled water, placing the reaction product in a vacuum drier at 70 ℃ for decompression to be dry, and grinding the reaction product and sieving the product by using a 300-mesh sieve to obtain the composite material (the composite montmorillonite polymer for short) of the polyethylene oxide and the clay.

The formula of the adsorption filler for the purification column is as follows: 70wt% of a composite material of polyethylene oxide and clay (abbreviated as composite montmorillonite polymer), 25wt% of activated silica gel and 5wt% of KH-900.

stirring the composite montmorillonite polymer, the activated silica gel and the silane coupling agent (KH-900) in a reaction kettle for 6 hours by a magnetic stirrer at the stirring speed of 150r/min to obtain a mixed solution; aging the mixed solution in 0.5mol/L sodium hydroxide solution for 4h to obtain microspheres, separating the microspheres from the solution, washing with washing solution for 5 times, and drying to obtain adsorption filler for the purification column, wherein the adsorption filler is marked as adsorption filler No. 10.

The activated silica gel was prepared in the same manner as in example 1.

Comparative example 1

This comparative example 1 is different from example 10 in that the sodium-modified montmorillonite, potassium-modified montmorillonite and calcified modified montmorillonite obtained in step 3) of example 10 were used as fillers directly.

Comparative example 2

The difference between this comparative example 2 and example 10 is that the formulation was made up of 70wt% of the composite montmorillonite polymer and 30wt% of the filler obtained in example 10 without adding the silane coupling agent KH-900.

Performance testing

After the adsorption filler for the purification columns of examples 1-10, the filler of comparative example 1 and the filler of comparative example 2 are used for purifying the matrix, the content of various mycotoxins in the matrix is determined by adopting an ultra-high performance liquid chromatography-mass spectrometry combined method, and the recovery rate is determined.

The matrix is selected from rice, vegetable oil, vinegar, chilli sauce, soybean, soy sauce, melon seeds, fermented soya beans, hawthorn slices, apples, hawthorn juice, etc.

1.1 measurement of the content of Aflatoxin B1, aflatoxin B2, aflatoxin G1 and Aflatoxin G2

1.1.1 preparation of solution

Standard stock solutions: accurately weighing a proper amount of standard substance, dissolving the standard substance with acetonitrile, and preparing into a standard stock solution of 100 mu g/mL.

Mixing standard intermediate liquid: each 100. Mu.L of the standard stock solution was accurately transferred, and the volume was adjusted to 10mL with acetonitrile, and the concentration was 1. Mu.g/mL.

Mixing standard working solution: accurately transferring 0.1mL of mixed standard intermediate solution, and diluting to 10mL with acetonitrile to a constant volume of 100ng/mL.

Acetonitrile-water solution (84 + 16): 840mL of acetonitrile is added to 160mL of water and mixed well.

1.1.2 extraction step

Weighing 5g of sample in a 50mL centrifuge tube, adding 20mL of acetonitrile-water solution (84 + 16), uniformly mixing in a vortex manner, placing in an ultrasonic/vortex oscillator or a shaking table for oscillation for 20min, centrifuging at 6000r/min for 10min, and taking the supernatant as a sample extracting solution for later use.

1.1.3, purifying step with multifunctional purifying column

1) 10mL of the sample extract obtained in step 1.1.2 was added to a glass tube.

2) A rubber head of a purification column containing the adsorbing filler for purification columns of examples 1 to 10, the filler of comparative example 1 and the filler of comparative example 2 was inserted into a test tube from the top end of the test tube, and the purification column was pressed down to the bottom end of the test tube to purify a sample extract.

3) And taking out the purified sample extracting solution to a centrifugal tube to obtain the purified extracting solution.

4) 5mL of purified extract was taken, blown dry with nitrogen, reconstituted with 1mL of the initial mobile phase (mobile phase conditions starting at 0min, say 68% of phase A plus 32% of phase B), vortexed for 30s to dissolve the residue, and the residue was analyzed on a millipore filter.

1.1.4, chromatography-Mass Spectrometry conditions

Ultra high performance liquid chromatography (UPLC) conditions

A chromatographic column: ultimate UHPLC XB-C18,1.8 μm, 2.1X 100mm

Protecting the column core: ultimate UHPLC XB-C18, ultra-high pressure column core 2.1X 5mm

Mobile phase: phase A is 5mmol/L ammonium acetate solution; phase B acetonitrile-methanol solution (50+50)

Column temperature: at 40 ℃; flow rate: 0.3mL/min; sample introduction volume: 1 μ L

Gradient elution procedure: see table 1.

TABLE 1 gradient elution procedure

Time/min	A phase/%)	B phase/%)
			0.00	68	32
0.50	68	32
			3.00	55	45
4.00	55	45
			4.20	0	100
4.80	0	100
			5.00	68	32
7.00	68	32

Conditions of Mass Spectrometry

Liquid instrument model: AB Sciex TRIPLE QUAD 4500

An ion source: electrospray ion source (ESI);

the scanning mode comprises the following steps: scanning positive ions;

the detection mode is as follows: multiple Reaction Monitoring (MRM) mode;

ion spray voltage: 5500V; ion source temperature: 500 ℃;

air curtain air (CUR) 10psi; atomizing gas (GS 1) 55psi; supplemental gas (GS 2) 50psi.

Other mass spectral parameters are shown in table 2.

TABLE 2 multiple reaction monitoring mode (MRM) parameters

1.2 content determination of zearalenone

1.2.1 preparation of solutions

Standard stock solutions: accurately weighing a proper amount of standard substance, dissolving the standard substance with acetonitrile, and preparing into standard stock solution with the concentration of 100 mu g/mL.

Standard working solution: accurately transferring 100 mu L of standard stock solution, and diluting to 10mL with acetonitrile, wherein the concentration is 1 mu g/mL.

Acetonitrile-water solution (9 + 1): 900mL of acetonitrile was added to 100mL of water and mixed well.

1.2.2 extraction step

Weighing 5g of sample in a 50mL centrifuge tube, adding 1g of sodium chloride, adding 20mL of acetonitrile-water solution (9 + 1), vortexing for 15min, centrifuging for 5min at 6000r/min, and taking the supernatant for later use.

1.2.3, purification step with multifunctional purification column

Same as step 1.1.3 in 1.1.

1.2.4 conditions of chromatography-Mass Spectrometry

Ultra high performance liquid chromatography (UPLC) conditions

A chromatographic column: ultimate UHPLC XB-C18,1.8 μm, 2.1X 100mm

Mobile phase: phase A is water; phase B of acetonitrile

Column temperature: 40 ℃; flow rate: 0.2mL/min; sample injection volume: 1 μ L

Gradient elution procedure: see table 3.

TABLE 3 gradient elution procedure

Conditions of Mass Spectrometry

Liquid quality instrument model: AB Sciex TRIPLE QUAD 4500

An ion source: electrospray ion source (ESI);

the scanning mode comprises the following steps: scanning negative ions;

the detection mode comprises the following steps: multiple Reaction Monitoring (MRM) mode;

ion spray voltage: -4500V; ion source temperature: 500 ℃;

curtain air (CUR) 10psi; atomizing gas (GS 1) 55psi; auxiliary gas (GS 2) 50psi.

Other mass spectral parameters are shown in table 4.

TABLE 4 multiple reaction monitoring mode (MRM) parameters

1.3 content determination of patulin

1.3.1 preparation of solutions

1.3.2 extraction step

Weighing 4g of sample in a 50mL centrifuge tube, adding 20mL of acetonitrile, performing ultrasonic treatment for 5min, centrifuging at 6000r/min for 5min, and taking supernatant as sample extracting solution for later use.

1.3.3, purification step with multifunctional purification column

1) 10mL of the sample extract obtained in step 1.3.2 was added to a glass tube, followed by 50. Mu.L of acetic acid.

2) A rubber head of the purification cartridge containing the adsorbing filler for purification cartridges of examples 1 to 10, the filler of comparative example 1 and the filler of comparative example 2 was inserted into a test tube from the top end of the test tube, and the purification cartridge was pressed down to the bottom end of the test tube to purify a sample extract.

3) And taking out the sample extracting solution purified at the upper part of the purifying column into a centrifugal tube to obtain the purified extracting solution.

4) 5mL of the purified extract was taken, 20. Mu.L of acetic acid was added, nitrogen was blown dry, redissolved with 1mL of the initial mobile phase (95% phase A and 5% phase B), the residue was vortexed for 30 seconds, and the cells were analyzed on a microfiltration membrane.

1.3.4 chromatographic conditions

Ultra high performance liquid chromatography (UPLC) conditions

A chromatographic column: ultimate UHPLC AQ-C18,1.8 μm, 2.1X 100mm

Protecting the column core: ultimate UHPLC AQ-C18, ultra-high pressure column core of 2.1 × 5mm

Mobile phase: a is water; b is acetonitrile

Column temperature: 30 ℃; flow rate: 0.3mL/min; sample introduction volume: 1 μ L

Gradient elution procedure: see table 5.

TABLE 5 gradient elution procedure

Conditions of Mass Spectrometry

Liquid instrument model: AB Sciex TRIPLE QUAD 4500

An ion source: electrospray ion source (ESI);

the scanning mode is as follows: scanning negative ions;

ion spray voltage: -4500V; ion source temperature: 500 ℃;

Other mass spectral parameters are shown in table 6.

TABLE 6 multiple reaction monitoring mode (MRM) parameters

FIG. 1 is a graph showing the results of examining the recovery rates of the toxins to be tested in the labeling extract solutions by using the adsorption packing for purification columns of examples 1 to 10 and the packing of comparative example 1 and comparative example 2.

The main components of the multifunctional purifying column of a certain domestic commercial brand 1 are silica gel and anion-cation polymer; reference 1, reference 1 is: high performance liquid chromatography measures vomitoxin content [ J ] in whole wheat flour, grain and oil food technology, 2018, 26 (5): 53-57.

The main components of the multifunctional purifying column of a certain commercial brand 2 in China are activated carbon, anionic and cationic polymer, copure 228.

As can be seen from FIG. 1, aflatoxin B was treated with the adsorbing fillers for purification columns of examples 1-10 ₁ (AFB ₁ ) Aflatoxin B ₂ (AFB ₂ ) Group, aflatoxins G ₁ (AFG ₁ ) Aflatoxin G ₂ (AFG ₂ ) Zearalenone (ZEN) and Patulin (PAT) all had good recovery rates and better universality, while comparative example 1 and comparative example 2 were less universal, and the recovery rates and universality of commercial purification column 1 and commercial purification column 2 were inferior to those of the examples, and had greater adsorption to each mycotoxin.

1.4 evaluation of matrix Effect

Evaluation was performed by an extraction and addition method, and a specific calculation method is shown in formula (1).

If ME (%) < 100%, indicating an inhibitory effect;

if ME (%) > 100%, indicating an enhancement effect;

it is reported in the literature that when the ME value is between 80 and 120%, it indicates that there is no significant matrix effect, the effect of which is negligible.

And (4) converting a standard curve established by an external standard method to evaluate the matrix effect.

The results of the above experiments and FIG. 1 were calculated to show that the ME values of examples 1 to 10 were between 80% and 120%. And the ME values of the comparative examples 1 and 2, the certain domestic commercial brand 1 and the certain domestic commercial brand 2 are not 80-120 percent, and obvious matrix effect exists.

1.5 evaluation of solvent Effect

The organic proportion of the redissolving solvent (the initial mobile phase) will have a varying degree of solvent effect on the analyte.

FIG. 2 shows the adsorption packing for purification column of example 10 and the packing of comparative examples 1 and 2 against aflatoxin B ₁ Aflatoxins B ₂ Aflatoxin G ₁ And aflatoxin G ₂ The solvent effect evaluation chart was performed. (Aflatoxin B from left to right ₁ Aflatoxins B ₂ Aflatoxin G ₁ And aflatoxin G ₂ )

As can be seen from fig. 2, the solution purified by the adsorption filler in the purification column of example 10 has no obvious solvent effect, while the solution in comparative example 1 and comparative example 2 and certain national commercial brand 1 and certain national commercial brand 2 have obvious interference of solvent effect.

1.6 accuracy, precision

Samples with negative or as low as possible mycotoxin content were weighed out and subjected to a three-level, six-parallel labeling experiment, and the results are shown in tables 7 and 8, in which the purification column after column packing was pretreated with the adsorption packing 10# for purification column of example 10 and the packing of comparative example 1, respectively.

Table 7 accuracy and precision results of example 10

Table 8 accuracy and precision results for comparative example 1

As is clear from tables 7 and 8, the recovery rate of 85% to 120% and the daytime precision of 1.2% to 5% using the adsorbent packing for purification column of example 10 are superior to those of comparative example 1.

1.7 filler morphology testing

FIG. 3 is a microscopic view of an adsorption packing for a purification column of example 10, a packing of comparative example 1, a packing of comparative example 2, a multifunctional purification column of a certain domestic commercial brand 1, and a multifunctional purification column of a certain domestic commercial brand 2.

As is clear from FIG. 3, the adsorbent filler for purification columns of example 10 had a uniform distribution of microspheres and a small impurity content (small irregularly shaped particles), and the particles having a small particle size were distributed in the range of 3 μm to 5 μm and the particles having a large particle size were distributed in the range of 10 μm to 15 μm.

1.8 batch reproducibility test of adsorption Filler for purification column

FIG. 4 is a baseline spectrum of a purification analysis of zearalenone using an adsorption packing for purification columns prepared in example 10 for the continuous synthesis of 6 batches.

As can be seen from FIG. 4, the peak points of the adsorption packing for 6 batches of purification columns almost completely overlapped.

The above results fully indicate that the adsorbing material of the present invention has good batch stability, and further indicate that the adsorbing filler for purification column used in the present study has excellent stability and reproducibility.

In conclusion, the adsorption filler for the purification column has the advantages that the microspheres with small particle size and large particle size are uniformly distributed, and the adsorption filler for the purification column can treat aflatoxin B ₁ Aflatoxins B ₂ Aflatoxin G ₁ Aflatoxin G ₂ The method has the advantages of large adsorption of various mycotoxins such as Zearalenone (ZEN) and Patulin (PAT), small matrix effect and solvent effect interference, high recovery rate, low RSD (soluble protein) and high stability, and is particularly suitable for mycotoxin detection.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Those skilled in the art can modify or change the above-described embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be made by those skilled in the art without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. The composite material of the polyethylene oxide and the clay is characterized by comprising the following raw materials in parts by weight:

4000-8000 parts of inorganic salt modified clay

50 to 100 portions of quaternary ammonium salt compound

1 part of polyethylene oxide.

2. The composite material according to claim 1, characterized by comprising at least one of the following technical features:

the inorganic salt modified clay is selected from one or more of sodium modified clay, potassium modified clay and calcification modified clay; preferably, the inorganic salt modified clay is a mixture of sodium modified clay, potassium modified clay and calcification modified clay; more preferably, the mass ratio of the sodium modified clay to the potassium modified clay to the calcification modified clay is (0.5-2): (0.5-2) 1;

the quaternary ammonium salt compound is selected from one or more of N, N, N-trimethyl-2- (2-methyl-1-oxo-2-propenyl oxy) ethyl ammonium chloride-acrylamide copolymer, chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether, alkyl dimethyl benzyl ammonium halide and dialkyl dimethyl ammonium halide;

the ratio of the weight average molecular weight to the number average molecular weight of the polyethylene oxide is 17-20, the particle size is 300-400 meshes, and the dissolving temperature is 30-50 ℃.

3. The composite material of claim 1, wherein the inorganic salt modified clay is prepared by a method comprising:

3) And (3) adjusting the pH value by adopting alkali, adding inorganic salt, mixing, centrifuging and drying to obtain the inorganic salt modified clay.

4. The composite material according to claim 3, characterized by comprising at least one of the following technical features:

1) Wherein the clay is one or more selected from kaolinite, montmorillonite, serpentine, talc and mica;

2) The oxidant is one or more selected from hydrogen peroxide, peracetic acid and nitric acid;

2) Wherein the pH value is 2-7;

3) Wherein the pH value is 8-10;

3) Wherein the inorganic salt is selected from one or more of sodium chloride, potassium chloride and calcium chloride;

the mass ratio of the clay to the inorganic salt is (5-7): (12-30);

the mass ratio of the clay to the oxidant is 1: (2 to 20).

5. A method for preparing a composite material according to any one of claims 1 to 4, characterized in that it comprises the following steps:

inorganic salt modified clay and quaternary ammonium salt compound react in solvent and then react with polyethylene oxide to obtain the composite material.

6. The method of claim 5, wherein the reaction temperature is 40 to 80 ℃;

and/or the polymerization reaction temperature is 20-40 ℃;

and/or, the solvent is selected from water.

7. Use of a composite material according to any one of claims 1 to 4 as a raw material in an adsorbent filler.

8. An adsorbent packing for a purification column, comprising the composite material according to any one of claims 1 to 4, an activated silica gel, and a silane coupling agent; preferably, the adsorption filler for the purification column comprises the following raw materials in parts by weight:

40 to 70 portions of composite material

20 to 50 portions of activated silica gel

5-12 parts of a silane coupling agent.

9. The adsorptive filler for purification columns according to claim 8, wherein said silane coupling agent is selected from one or more of γ -aminopropyltriethoxysilane, vinyltriethoxysilane, N- (β -aminoethyl) - γ -aminopropyl-tri (ethyl) oxysilane, γ -aminoethylaminopropyl-trimethoxysilane;

and/or the preparation method of the activated silica gel comprises the following steps: mixing the silica gel raw powder with acid, and drying to obtain the activated silica gel.

10. The method for preparing an adsorptive filler for a purification column according to claim 8 or 9, comprising the steps of:

and mixing the raw material components, and aging to obtain the adsorption filler for the purification column.

11. Use of the adsorption packing for purification columns according to claim 8 or 9 as chromatography material.