CN108872447A - The detection method of thyroid imhibitor based on hydrophily Solid Phase Extraction - Google Patents
The detection method of thyroid imhibitor based on hydrophily Solid Phase Extraction Download PDFInfo
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Abstract
A kind of detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, prepares the solid-phase extraction column of the amine-modified hybridisation silica gel of polyethyleneimine and with acetonitrile rinse first, deionized water rinse, then with 98%（v/v）Acetonitrile solution balance elutes solid-phase extraction column with pure acetonitrile by sample to be tested by above-mentioned solid-phase extraction column, then with containing 50%（v/v）The ammonium acetate solution of the pH 2 of acetonitrile elutes solid-phase extraction column, and the pH tune of eluent causes neutrality, after filtering, makees hydrophilic chromatographic separation and detection.The hydrophily solid phase extraction method that the present invention establishes, 15.8 mg/g are up to the adsorption capacity of thyroid imhibitor, and loading volume reaches 40 mL, acetonitrile extracting solution loading can be directly used, eluent directly makees chromatography with hydrophilic, and the dry and molten step of weight is omitted, simplify sample pre-treatments step, sample pre-treatments for thyroid imhibitor in complex matrices for the first time improve the accuracy of method, reduce detection error.
Description
Technical field
The present invention relates to sample pretreatment drug detection technique field, especially a kind of first based on hydrophily Solid Phase Extraction
The detection method of shape gland inhibitor.
Background technique
Thyroid imhibitor is a kind of polar thioamide analog compound for inhibiting thyroid function, for example thiocarbamide is phonetic
Pyridine, methylthiouracil, propylthiouracil and phenylthiourea pyrimidine etc..After feeding animal, gastrointestinal contents increasing can lead to
More, water content increases in internal organ and musculature.Such drug, which is illegally added to play in animal feed, obviously increases weight
Effect.If eating these medicament residues after animal tissue to be absorbed by the body, teratogenesis carcinogenesis can be generated.Therefore, meat is eaten
The detection of thyroid imhibitor is extremely important in product.
Due to thyroid imhibitor intrinsic polarity and hydrophily, it is caused to be difficult to extract from complicated matrix.
Most common extracting solution is methanol and acetonitrile.Due to the complexity of matrix, crude extract must use silica gel, C18 or ion
Exchange Solid Phase Extraction column purification.Wherein, silica gel solid-phase extraction column is suitable for the analysis of polarity and hydrophilic compounds, is widely applied
The purification of Thyreoidine inhibitor.Since extraction mechanism is normal-phase chromatography retention mechanism, methanol and acetonitrile are not suitable for direct loading
Onto silica gel solid-phase extraction column, weight is molten into the lower solvent of polarity after extracting solution must be dried, such as methylene chloride and ring
Hexane.In addition, lower detection limit, eluent also need drying in order to obtain, lay equal stress on molten to being suitable for liquid chromatogram or gas phase
In chromatographic isolation and the solution of detection.Therefore, purification process based on silica gel solid-phase extraction column not only complex steps, and it is time-consuming
Arduously, it is easy to produce error.
Hydrophily solid phase extraction method is the sample pre-treatments skill for being based primarily upon hydrophilic interaction mechanism having recently been developed
Art, the object of analysis are polarity and hydrophilic compounds.Hydrophily solid phase extraction method is widely used in Phosphorylated Peptide
Selective enrichment, adsorbent are the key that establish solid phase extraction method.Currently, the adsorbent for being applied to hydrophily Solid Phase Extraction is deposited
In the small disadvantage of loading volume, such as silica gel [B.D. Cai, J.X. Zhu, Z.G. Shi, B.F. Yuan, Y.Q.
Feng, J. Chromatogr. B 2013,942-943,31-36], pollen grain [Q. Lu, J.H. Wu, Q.W. Yu,
Y.Q. Feng, J. Chromatogr. A 2014,1367,39-47] and ampholytes modification hybridization silica gel material
[T.T. Wang, Y.M. Zhu, J.F. Ma, R.R. Xuan, H.Q. Gao, Z. Liang, L.H. Zhang,
Y.K. Zhang, J. Sep. Sci. 2015,38,87-92] loading volume be respectively less than 1.5 mL.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of hybridization silica gel material amine-modified using polyethyleneimine as inhale
Attached dose of solid-phase extraction column, purified treatment and detection method for thyroid imhibitor in animal tissue.
Above-mentioned purpose is implemented with the following technical solutions in the present invention：
The detection method of thyroid imhibitor based on hydrophily Solid Phase Extraction, feature include the following steps：
A, the solid-phase extraction column for the hybridization silica gel material that filled polyethylene imines is modified first is used into acetonitrile rinse, then uses deionized water
Rinse finally balances solid-phase extraction column with 98% acetonitrile；
B, sample to be tested is discarded useless by walking the solid-phase extraction column of rinse and balance through a with pure acetonitrile elution solid-phase extraction column
Liquid；
C, the solid-phase extraction column that pure acetonitrile elution will be walked by b, elutes solid-phase extraction column with low-concentration acetonitrile solution, collects elution
Liquid adjusts pH and causes neutral, use to be analyzed；
D, the eluent Jing Guo step c is separated and is detected using hydrophilic Interaction Chromatography.
The detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, it is characterized in that the polyethyleneimine
The preparation method of amine-modified hybridization silica gel material：The side that the hybridization silica gel material that amido is contained on surface is coupled by glutaraldehyde
Method is amine-modified on hybridization silica gel material by polyethyleneimine.
The detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, it is characterized in that the Solid Phase Extraction
The production method of column：Amine-modified 0.1~1.0 g of hybridization silica gel material of polyethyleneimine is weighed, loading aperture is 5 μm of lower sieve plates
1~10 mL Solid Phase Extraction empty tube column in, being loaded on aperture is 20 μm of upper sieve plate, and compacting is stand-by.
The detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, it is characterized in that hydrophily solid phase extracts
Take method：It with high concentration acetonitrile loading, is eluted with low-concentration acetonitrile, the aqueous solution that loading acetonitrile concentration is 90%~98%, elution
Acetonitrile concentration is the ammonium acetate solution of 10%~70% and pH 2.
The detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, it is characterized in that the hydrophilic interaction
The method of chromatographic isolation and detection：Chromatographic column is nh 2 column, and mobile phase is 95% acetonitrile solution, 0.8 mL/min of flow velocity, sample introduction
20 μ L, 30 °C of column temperature, 270 nm of wavelength.
The measuring method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, it is characterized in that being pressed down with Thyreoidine
The detection method of preparation：By the solid-phase extraction column for the hybridization silica gel material that filled polyethylene imines is modified use respectively 5 mL acetonitriles and
5 mL deionized water rinses, then solid-phase extraction column is balanced with 5 mL, 98% acetonitrile；Then it will contain the to be measured of thyroid imhibitor
1~40 mL of sample crosses column, elutes solid-phase extraction column with the pure acetonitrile of 2 mL, discards waste liquid；Contain the pH 2 of 50% acetonitrile with 1 mL again
10 mmol/L ammonium acetates elute solid-phase extraction column, collect eluent；Finally, being adjusted in the pH cause of eluent with sodium bicarbonate
Property, after crossing 0.45 μm of filter membrane, detected to hydrophilic Interaction Chromatography.
The detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, it is characterized in that being used for animal tissue
The detection method of thyroid imhibitor：The solid-phase extraction column for the hybridization silica gel material that filled polyethylene imines is modified is used 5 respectively
ML acetonitrile and 5 mL deionized water rinses, then solid-phase extraction column is balanced with 5 mL, 98% acetonitrile；Then extracting solution is used 10 respectively
ML and 40 mL cross column, elute solid-phase extraction column with the pure acetonitrile of 2 mL, discard waste liquid；10 of pH 2 with 1 mL containing 50% acetonitrile again
Mmol/L ammonium acetate elutes solid-phase extraction column, collects eluent；Finally, causing neutral, mistake with the pH that sodium bicarbonate adjusts eluent
After 0.45 μm of filter membrane, detected to hydrophilic Interaction Chromatography.
Advantages of the present invention and achievement are：Polyethyleneimine is prepared polyethyleneimine by the method that glutaraldehyde is coupled to repair
The hybridization silica gel material of decorations establishes hydrophily solid phase extraction method using the hydrophily of polyethyleneimine, is applied to for the first time multiple
The sample pretreatment of thyroid imhibitor in miscellaneous matrix.The amine-modified hybridization silica gel material of polyethyleneimine is to thyroid imhibitor
Adsorption capacity is up to 15.8 mg/g, the significantly larger than hybridization silica gel material of ampholytes modification（30 μ g/g, i.e. 0.03 mg/
g）, and maximum loading volume reaches 40 mL, higher than the method for the hydrophily Solid Phase Extraction polar micromolecules reported before.This
Invent the hydrophily solid phase extraction method for the hybridization silica gel material amine-modified based on polyethyleneimine established, directly mentioning with acetonitrile
Liquid loading is taken, eluent is directly analyzed with hydrophilic Interaction Chromatography, and step dry twice and that weight is molten is omitted, enormously simplifies sample
Product pre-treatment step improves the accuracy of method, reduces error.
Detailed description of the invention
Fig. 1 is hybridization silica gel material（A）The amine-modified hybridization silica gel material with polyethyleneimine（B）Scanning electron microscope (SEM) photograph；
The adsorption rate and loading acetonitrile concentration variation diagram of Fig. 2 thyroid imhibitor；
The rate of recovery and elution acetonitrile concentration variation diagram of Fig. 3 thyroid imhibitor；
The rate of recovery and pH variation diagram of Fig. 4 thyroid imhibitor；
The rate of recovery and loading volume variation diagram of Fig. 5 thyroid imhibitor；
The rate of recovery and elution volume variation diagram of Fig. 6 thyroid imhibitor；
The adsorption capacity and initial concentration variation diagram of Fig. 7 deracil.
Specific embodiment
The invention will be further described with reference to embodiments.
Embodiment 1：The solid-phase extraction column based on the amine-modified hybridization silica gel material of polyethyleneimine is made according to the following steps.
（1）The preparation of the amine-modified hybridization silica gel material of polyethyleneimine
In 50 mL plastic centrifuge tubes, 160 mg cetyl trimethylammonium bromides are weighed, 2.24 mL tetraethoxy-silicanes are added
Alkane, 2.36 mL aminopropyl triethoxysilanes, 4.35 mL dehydrated alcohols add 0.64 mL water after mixing dissolution, mix
Afterwards, 40 °C of water-baths are immediately placed in and react 24 h.After the reaction was completed, respectively with 100 mmol/L phosphorus of 15 mL methanol and 15 mL
Hydrochlorate（pH 8.0）It rinses 2~3 times, adds 2 mL 10%（v/v）It is dissolved in 100 mmol/L phosphate（pH 8.0）Penta 2
Aldehyde solution, 90 °C of 0.5 h of reaction, it can be observed that color becomes brown, then with 15 mL, 100 mmol/L phosphate（pH
8.0）After flushing, 0.625 mL is added and is dissolved in 100 mmol/L phosphate（pH 8.0）0.05 g/mL polyethyleneimine it is molten
Liquid and 5 mg/mL NaCNBH3, after 90 °C of 0.5 h of reaction, after being rinsed respectively with 15 mL distilled water and 15 mL methanol, 60
°C drying 10 h it is stand-by.After the material of above-mentioned drying is ground, wait load.Scanning electron microscope such as Fig. 1（A）Shown, diameter is less than 200
The sphere interconnected of nm constitutes porous structure, can provide high-specific surface area for the modification of polyethyleneimine.It is interconnected
Spherome surface is significantly covered with layer of material, shows that polyethyleneimine is successfully modified on hybridization silica gel material（Figure
1B）.
（2）The filling of solid-phase extraction column
Weigh step（1）Amine-modified 0.18 g of hybridization silica gel material of the polyethyleneimine of processing is packed into sieve plate under 5 μm of apertures of band
3 mL solid-phase extraction columns in, after inserting 20 μm of aperture upper sieve plates, compacting is stand-by.
Embodiment 2：Using three kinds of thyroid imhibitor standard items as sample, loading acetonitrile concentration is optimized.
Using the solid-phase extraction column based on the amine-modified hybridization silica gel material of polyethyleneimine made by embodiment 1, respectively
With 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL 98%（v/v）Acetonitrile solution balances solid-phase extraction column, by three kinds
The standard items of thyroid imhibitor are dissolved in 60%, 70%, 80%, 85%, 90%, 95% and 98%（v/v）In acetonitrile solution, concentration
For 2.0 μ g/mL, 1 mL of loading, the sample of outflow is collected, is analyzed after crossing 0.45 μm of filter membrane with hydrophilic Interaction Chromatography.Chromatography point
Analysis condition：Analytical column is Shimadzu Inertsil-NH2（4.6×150 mm, 3 μm）, mobile phase 95%（v/v）Aqueous acetonitrile
Liquid, 0.8 mL/min of flow velocity, sample introduction 20 μ L, 30 °C of column temperature, 270 nm of Detection wavelength.As a result as shown in Fig. 2, adsorption rate with
The increase of acetonitrile concentration and increase, when acetonitrile concentration be greater than 90% when, adsorption rate reaches maximum value, that is, illustrates that sample is inhaled completely
It is attached on solid-phase extraction column.Therefore, the concentration of loading acetonitrile can be 90%~98%, and the present invention uses 98%（v/v）Aqueous acetonitrile
Liquid dissolves as sample or extracts solution.
Embodiment 3：Using three kinds of thyroid imhibitor standard items as sample, elution acetonitrile concentration is optimized.
Using the solid-phase extraction column based on the amine-modified hybridization silica gel material of polyethyleneimine made by embodiment 1, respectively
With 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL 98%（v/v）Acetonitrile solution balances solid-phase extraction column, by three kinds
The standard items of thyroid imhibitor are dissolved in 98%（v/v）In acetonitrile solution, concentration is 2.0 μ g/mL, 1 mL of loading, with 2
After the pure acetonitrile elution of mL, with 1 mL 50%, 60%, 70%, 80% and 90%（v/v）Acetonitrile solution is eluted, and elution is collected
Liquid is detected after crossing 0.45 μm of filter membrane with hydrophilic Interaction Chromatography.Chromatographiccondition：Analytical column is Shimadzu Inertsil-NH2
（4.6×150 mm, 3 μm）, mobile phase 95%（v/v）Acetonitrile solution, 0.8 mL/min of flow velocity, 20 μ L of sample introduction, column temperature
30 °C, 270 nm of Detection wavelength.As a result as shown in figure 3, the rate of recovery is reduced with the increase of acetonitrile concentration, work as aqueous acetonitrile
Liquid concentration is 50%（v/v）When, the rate of recovery reaches maximum value.Therefore, using 50%（v/v）Acetonitrile solution is as eluent.
Embodiment 4：Using three kinds of thyroid imhibitor standard items as sample, the pH of elution solution is optimized.
Using the solid-phase extraction column based on the amine-modified hybridization silica gel material of polyethyleneimine made by embodiment 1, respectively
With 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL 98%（v/v）Acetonitrile solution balances solid-phase extraction column, by three kinds
The standard items of thyroid imhibitor are dissolved in 98%（v/v）In acetonitrile solution, concentration is 2.0 μ g/mL, 1 mL of loading, with 2
The pure acetonitrile of mL elutes and then contains 50% with 1 mL respectively（v/v）The 10 mmol/L second that the pH of acetonitrile is 2,3,4,5,6 and 8
Acid ammonium solution is eluted, and eluent is collected, and sodium bicarbonate is added or second acid for adjusting pH causes neutrality, after crossing 0.45 μm of filter membrane
It is detected for hydrophilic Interaction Chromatography.Chromatographiccondition：Analytical column is Shimadzu Inertsil-NH2（4.6×150 mm, 3 μm）,
Mobile phase is 95%（v/v）Acetonitrile solution, 0.8 mL/min of flow velocity, sample introduction 20 μ L, 30 °C of column temperature, 270 nm of Detection wavelength.
As a result as shown in figure 4, the rate of recovery is reduced with the increase of pH value, after reaching minimum value, increase with the increase of pH value.
When pH is 2, the rate of recovery reaches maximum value.Therefore, using 50%（v/v）Acetonitrile/10 mmol/L ammonium acetates（pH 2）As washing
De- liquid.
Embodiment 5：Using three kinds of thyroid imhibitor standard items as sample, loading volume is optimized.
Using the solid-phase extraction column based on the amine-modified hybridization silica gel material of polyethyleneimine made by embodiment 1, respectively
With 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL 98%（v/v）Acetonitrile solution balances solid-phase extraction column, keeps three
Each 2.0 μ g of standard items of kind thyroid imhibitor is constant, with 98%（v/v）Acetonitrile solution is diluted to 1,5,10,20,40 and 80
ML, respectively by solid-phase extraction column, with the pure acetonitrile elution of 2 mL and then respectively with 1 mL 50%（v/v）Acetonitrile/10 mmol/
L ammonium acetate（pH 2）It is eluted, collects eluent, sodium bicarbonate is added and adjusts pH cause neutrality, crosses after 0.45 μm of filter membrane for parent
The detection of water action chromatography.Chromatographiccondition：Analytical column is Shimadzu Inertsil-NH2（4.6×150 mm, 3 μm）, flowing
It is mutually 95%（v/v）Acetonitrile solution, 0.8 mL/min of flow velocity, sample introduction 20 μ L, 30 °C of column temperature, 270 nm of Detection wavelength.As a result
As shown in figure 5, loading volume is very big on rate of recovery influence, the maximum loading volume of propylthiouracil and methylthiouracil is
10 mL, and the maximum loading volume of deracil reaches 40 mL, i.e. propylthiouracil and methylthiouracil run through body
Product is 10 mL, and the volume that runs through of deracil is 40 mL.Therefore, the loading volume of propylthiouracil and methylthiouracil
10 mL are selected, the loading volume of deracil selects 40 mL.
Embodiment 6：Using three kinds of thyroid imhibitor standard items as sample, elution volume is optimized.
Using the solid-phase extraction column based on the amine-modified hybridization silica gel material of polyethyleneimine made by embodiment 1, respectively
With 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL 98%（v/v）Acetonitrile solution balances solid-phase extraction column, by three kinds
The standard items of thyroid imhibitor are dissolved in 98%（v/v）In acetonitrile solution, concentration is 2.0 μ g/mL, 1 mL of loading, with 2
The pure acetonitrile of mL elutes and then respectively with 0.5,0.75,1,1.5 and 2 mL 50%（v/v）Acetonitrile/10 mmol/L ammonium acetates（pH
2）It is eluted, collects eluent, sodium bicarbonate is added and adjusts pH cause neutrality, crosses after 0.45 μm of filter membrane for hydrophilic Interaction Chromatography
Detection.Chromatographiccondition：Analytical column is Shimadzu Inertsil-NH2（4.6×150 mm, 3 μm）, mobile phase 95%（v/
v）Acetonitrile solution, 0.8 mL/min of flow velocity, sample introduction 20 μ L, 30 °C of column temperature, 270 nm of Detection wavelength.As a result as shown in fig. 6,
The rate of recovery increases with the increase of elution volume, and when elution volume is greater than 1 mL, the rate of recovery reaches maximum value and keeps not
Become.Therefore, minimum detection limit, elution volume select 1 mL in order to obtain.
Embodiment 7：Using deracil as sample, absorption of the amine-modified hybridization silica gel material of polyethyleneimine to it is determined
Capacity.
Deracil is dissolved in 40 mL（V）98%（v/v）In acetonitrile solution, initial concentration is in 3 to 121 mg/L
（C0）Between, it is then loaded to and is filled with 0.23 g（m）In the solid-phase extraction column of the amine-modified hybridization silica gel material of polyethyleneimine,
It is C with the concentration that hydrophilic Interaction Chromatography detects after collecting effluxt, adsorption capacity is calculated according to following formula：Qe=(C0-
Ct)V/m.Relationship between adsorption capacity and initial concentration is as shown in fig. 7, there it can be seen that adsorption capacity is dense with originating
The increase of degree and increase, when initial concentration be greater than 109 mg/L when, adsorption capacity reaches maximum value and remains unchanged.Therefore, gather
The hybridization silica gel material of aziridine modification is 15.8 mg/g to the maximum adsorption capacity of deracil, significantly larger than both sexes electricity
Solve adsorbance of the hybridization silica gel material to melamine of matter modification（30 μg/g）.
Concrete application embodiment：
Embodiment 8：In order to verify the solid phase based on the amine-modified hybridization silica gel material of polyethyleneimine made by filling embodiment 1
Extraction column is to the selective adsorption capacity of thyroid imhibitor, by the solid-phase extraction column of brand-new to thyroid imhibitor in pork liver
Extracting solution does Solid phase extraction processing, then carries out analysis measurement to thyroid imhibitor with hydrophilic Interaction Chromatography.
The preparation of pork liver extracting solution：After the homogenate of 25 g pork livers, 30 mL 98% are added（v/v）In acetonitrile solution, it is added three
Kind thyroid imhibitor standard items make 5 μ g/kg of deracil concentration, propylthiouracil and methylthiouracil concentration point
Not Wei 20 μ g/kg, after balancing 30 min, ultrasonic extraction 10 min, subsequent 10000 rpm is centrifuged 10 min, and extracting solution is shifted
Into another centrifuge tube, 20 mL 98% are added in pork liver slurry after extraction（v/v）Acetonitrile solution, ultrasonic extraction
After 10 min, 10000 rpm are centrifuged 10 min, finally, merging two kinds of extracting solutions, for use.
Extraction conditions：Using the solid phase based on the amine-modified hybridization silica gel material of polyethyleneimine made by filling embodiment 1
Extraction column, respectively with 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL 98%（v/v）Acetonitrile solution balances solid phase extraction
Take column；Then extracting solution is crossed into column with 10 mL and 40 mL respectively, elutes solid-phase extraction column with the pure acetonitrile of 2 mL, discards waste liquid；
Again with 1 mL 50%（v/v）Acetonitrile/10 mmol/L ammonium acetates（pH 2）Solid-phase extraction column is eluted, eluent is collected；Finally, with
The pH that sodium bicarbonate adjusts eluent causes neutrality, after crossing 0.45 μm of filter membrane, detects to hydrophilic Interaction Chromatography.
The result shows that the spiked levels of propylthiouracil and methylthiouracil are respectively 20 in pork liver extracting solution
The spiked levels of μ g/kg, deracil are 5 μ g/kg, and the rate of recovery is respectively 94.5%, 100.4% and 100.8%, and RSD is respectively
9.6%, 6.9% and 4.5%, this method is suitble to the detection and analysis of the thyroid imhibitor in animal tissue.
Embodiment 9：The deracil concentration being added in the extracting solution of pork liver is 100 μ g/kg, propylthiouracil and first
Base deracil concentration is respectively 400 μ g/kg, remaining condition is the same as embodiment 8.The result shows that in pork liver extracting solution, propyl
The spiked levels of deracil and methylthiouracil are respectively 400 μ g/kg, and the spiked levels of deracil are 100 μ g/
Kg, the rate of recovery are respectively 96.0%, 102.8% and 97.7%, and RSD is respectively 5.4%, 3.6% and 7.7%, and this method is suitble to animal groups
The detection and analysis of thyroid imhibitor in knitting.
Embodiment 10：In order to further verify the hydrophily solid phase extraction method established by the present invention that is based on to other first shapes
The measurement effect of gland inhibitor, the phenylthiourea pyrimidine concentration being added in the extracting solution of pork liver are 20 μ g/kg, remaining condition
With embodiment 8.The result shows that the spiked levels of phenylthiourea pyrimidine are 20 μ g/kg, and the rate of recovery is in pork liver extracting solution
93.7%, RSD 6.5%, this method are suitble to the detection and analysis of the thyroid imhibitor in animal tissue.
Claims (7)
1. the detection method of the thyroid imhibitor based on hydrophily Solid Phase Extraction, feature include the following steps：
A, the solid-phase extraction column for the hybridization silica gel material that filled polyethylene imines is modified first is used into acetonitrile rinse, then uses deionized water
Rinse finally balances solid-phase extraction column with 98% acetonitrile；
B, sample to be tested is discarded useless by walking the solid-phase extraction column of rinse and balance through a with pure acetonitrile elution solid-phase extraction column
Liquid；
C, the solid-phase extraction column that pure acetonitrile elution will be walked by b, elutes solid-phase extraction column with low-concentration acetonitrile solution, collects elution
Liquid adjusts pH and causes neutral, use to be analyzed；
D, the eluent Jing Guo step c is separated and is detected using hydrophilic Interaction Chromatography.
2. the detection method of the thyroid imhibitor according to claim 1 based on hydrophily Solid Phase Extraction, it is characterized in that
The preparation method of the amine-modified hybridization silica gel material of the polyethyleneimine：The hybridization silica gel material of amido is contained by penta in surface
The method of dialdehyde coupling is amine-modified on hybridization silica gel material by polyethyleneimine.
3. the detection method of the thyroid imhibitor according to claim 1 based on hydrophily Solid Phase Extraction, it is characterized in that
The production method of the solid-phase extraction column：Amine-modified 0.1~1.0 g of hybridization silica gel material of polyethyleneimine is weighed, aperture is packed into
In 1~10 mL Solid Phase Extraction empty tube column for 5 μm of lower sieve plates, it is loaded on the upper sieve plate that aperture is 20 μm, compacting is stand-by.
4. the detection method of the thyroid imhibitor according to claim 1 based on hydrophily Solid Phase Extraction, it is characterized in that
Hydrophily solid phase extraction method：It with high concentration acetonitrile loading, is eluted with low-concentration acetonitrile, loading acetonitrile concentration is 90%~98%
Aqueous solution, the ammonium acetate solution that elution acetonitrile concentration is 10%~70% and pH 2.
5. the detection method of the thyroid imhibitor according to claim 1 based on hydrophily Solid Phase Extraction, it is characterized in that
The method of the hydrophilic Interaction Chromatography separation and detection：Chromatographic column is nh 2 column, and mobile phase is 95% acetonitrile solution, flow velocity 0.8
ML/min, sample introduction 20 μ L, 30 °C of column temperature, 270 nm of wavelength.
6. the detection method of the thyroid imhibitor according to claim 1 based on hydrophily Solid Phase Extraction, it is characterized in that
Detection method for thyroid imhibitor：The solid-phase extraction column for the hybridization silica gel material that filled polyethylene imines is modified is distinguished
Solid-phase extraction column is balanced with 5 mL acetonitriles and 5 mL deionized water rinses, then with 5 mL, 98% acetonitrile；Then it will contain thyroid gland
The sample to be tested of inhibitor crosses column with 1~40 mL respectively, elutes solid-phase extraction column with the pure acetonitrile of 2 mL, discards waste liquid；Again with 1
The 10 mmol/L ammonium acetates of pH 2 of the mL containing 50% acetonitrile elute solid-phase extraction column, collect eluent；Finally, with sodium bicarbonate tune
The pH for saving eluent causes neutrality, after crossing 0.45 μm of filter membrane, detects to hydrophilic Interaction Chromatography.
7. the detection method of the thyroid imhibitor according to claim 6 based on hydrophily Solid Phase Extraction, it is characterized in that
Detection method for animal tissue's thyroid imhibitor：The solid phase for the hybridization silica gel material that filled polyethylene imines is modified extracts
It takes column to use 5 mL acetonitriles and 5 mL deionized water rinses respectively, then balances solid-phase extraction column with 5 mL, 98% acetonitrile；Then it will mention
It takes liquid to cross column with 10mL and 40 mL respectively, elutes solid-phase extraction column with the pure acetonitrile of 2 mL, discard waste liquid；Contain 50% second with 1 mL again
The 10 mmol/L ammonium acetates of the pH 2 of nitrile elute solid-phase extraction column, collect eluent；Finally, adjusting eluent with sodium bicarbonate
PH cause neutral, after crossing 0.45 μm of filter membrane, detected to hydrophilic Interaction Chromatography.
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| CN201811041739.1A CN108872447B (en) | 2018-09-07 | 2018-09-07 | Hydrophilic solid phase extraction-based thyroid inhibitor detection method |
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| CN111495337A (en) * | 2020-04-22 | 2020-08-07 | 宁波工程学院 | Adsorbent for extracting fluoroquinolone medicines based on hydrophilicity and preparation method and extraction method thereof |
| CN111921509A (en) * | 2020-07-16 | 2020-11-13 | 深圳职业技术学院 | Solid phase extraction column and preparation method thereof and lactoferrin detection method |
| CN115166115A (en) * | 2022-07-20 | 2022-10-11 | 宁波工程学院 | Sample pretreatment method for detecting various mercury morphological contents in fish meat and detection method |
| CN115290806A (en) * | 2022-08-01 | 2022-11-04 | 宁波工程学院 | Method for solid-phase extraction of biogenic amine through hydrophilicity and cation exchange |
| WO2023129732A1 (en) * | 2021-12-31 | 2023-07-06 | Laboratory Corporation Of America Holdings | Methods and systems for hydrophilic-phase extraction |
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| CN111495337A (en) * | 2020-04-22 | 2020-08-07 | 宁波工程学院 | Adsorbent for extracting fluoroquinolone medicines based on hydrophilicity and preparation method and extraction method thereof |
| CN111495337B (en) * | 2020-04-22 | 2023-08-11 | 宁波工程学院 | Adsorbent for hydrophilic extraction of fluoroquinolone drugs, preparation method and extraction method thereof |
| CN111921509A (en) * | 2020-07-16 | 2020-11-13 | 深圳职业技术学院 | Solid phase extraction column and preparation method thereof and lactoferrin detection method |
| CN111921509B (en) * | 2020-07-16 | 2021-04-20 | 深圳职业技术学院 | Solid phase extraction column and preparation method thereof and lactoferrin detection method |
| WO2023129732A1 (en) * | 2021-12-31 | 2023-07-06 | Laboratory Corporation Of America Holdings | Methods and systems for hydrophilic-phase extraction |
| CN115166115A (en) * | 2022-07-20 | 2022-10-11 | 宁波工程学院 | Sample pretreatment method for detecting various mercury morphological contents in fish meat and detection method |
| CN115290806A (en) * | 2022-08-01 | 2022-11-04 | 宁波工程学院 | Method for solid-phase extraction of biogenic amine through hydrophilicity and cation exchange |
| CN115290806B (en) * | 2022-08-01 | 2024-05-28 | 宁波工程学院 | Method for solid phase extraction of biogenic amine by hydrophilic effect and cation exchange |
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