CN105384872A - Preparation method and use of penicillin G sodium salt surface molecular imprinting polymer - Google Patents
Preparation method and use of penicillin G sodium salt surface molecular imprinting polymer Download PDFInfo
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- CN105384872A CN105384872A CN201510934634.9A CN201510934634A CN105384872A CN 105384872 A CN105384872 A CN 105384872A CN 201510934634 A CN201510934634 A CN 201510934634A CN 105384872 A CN105384872 A CN 105384872A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F222/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical and containing at least one other carboxyl radical in the molecule; Salts, anhydrides, esters, amides, imides, or nitriles thereof
- C08F222/10—Esters
- C08F222/12—Esters of phenols or saturated alcohols
- C08F222/14—Esters having no free carboxylic acid groups, e.g. dialkyl maleates or fumarates
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/268—Polymers created by use of a template, e.g. molecularly imprinted polymers
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
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- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/26—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2335/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical, and containing at least one other carboxyl radical in the molecule, or of salts, anhydrides, esters, amides, imides or nitriles thereof; Derivatives of such polymers
- C08J2335/02—Characterised by the use of homopolymers or copolymers of esters
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Abstract
The invention discloses a preparation method of a penicillin G sodium salt surface molecular imprinting polymer. The preparation method comprises dissolving penicillin G sodium salt in a solvent which is a mixture of dimethyl sulfoxide and methanol according to a volume ratio of 4: 1, adding a functional monomer into the solution, carrying out stirring, adding a cross-linking agent and an initiator into the mixed solution, carrying out nitrogen deoxygenation, carrying out a heating reaction process, carrying out washing to remove penicillin G sodium salt as a template molecule after the reaction and carrying out drying to obtain the penicillin G sodium salt surface molecular imprinting polymer. The preparation method has simple processes, good operationality and a low preparation cost. The penicillin G sodium salt surface molecular imprinting polymer has a fast penicillin sodium salt molecule bonding rate, good specificity, high bonding capacity and short adsorption balance time. The penicillin G sodium salt surface molecular imprinting polymer has high desired sample selectivity, good specificity and recovery and recycle of a template molecule and can be used for detecting a certain concentration of penicillin G sodium salt residues in milk.
Description
Technical field
The invention belongs to molecular engram material technical field, particularly relate to a kind of preparation method and application thereof of Benzylpenicillin sodium molecular imprinted polymer on surface.
Background technology
Benzylpenicillin sodium is β-lactam antibitics the earliest, has stronger anti-microbial effect to gram-positive microorganism and some Gram-negative bacteria.Penicillin can destroy the cell walls of bacterium and play germicidal action in the nursery stage of bacterial cell, has good dispersiveness, hypotoxicity in vivo, and the efficiency that treatment is infected is high, in clinical treatment, have vital role.Along with Penicillin antibiotics is in the generally application of livestock industry, causes the microbiotic such as penicillin remaining in food, certain harm is caused to human body.The milk that such as edible residual penicillin is too much can make groups of people produce irritated effect, and the normal microflora of enteron aisle can be caused to lack of proper care, and causes other diseases related, and pathogenic bacteria in body is developed immunity to drugs.In addition, the growth of the microbiotic meeting lactic acid bacteria inhibiting contained in the milk of diary processing, therefore unfavorable to the processing of Yoghourt and cheese.
At present, the mensuration of the β-lactam antibiticss such as PEN-G (penicillin G) mainly contains following several method: (1) microbial method, as chlorinated triphenyl base triazole method (TTC method), paper disk method (PD), detection method (SP method) is irrigated by Dell, adenosine triphosphate method, enzymic colorimetric, cylinder plate method etc.; (2) immunoassay, as Enzyme-Linked Immunospot, immunosensor, fluorescence immunoassay (FIA); (3) physical and chemical inspection method, as liquid phase chromatography, liquid chromatography-mass spectrography, vapor-phase chromatography, tlc, capillary electrophoresis, hexavalent chrome bio-removal, gel electrophoresis-bioautography, fluorimetry etc.In biological sample, the content of PEN-G is lower, the validity of example enrichment purifying is directly connected to the validity of analytical procedure, traditional liquid-liquid, solid-liquid are difficult to realize the enrichment to trace P EN-G in biological sample and purifying, are difficult to carry out qualitative and quantitative analysis to PEN-G in sample; Biological material need be processed into the form being easy to analyze by chromatography and capillary electrophoresis etc., and enzyme-linked immunosorbent assay accuracy is inadequate, often occurs false positive event, poor anti jamming capability.
Molecular imprinting (MolecularImprintingTechnology, MIT) refers to a kind of new technology that can be used for synthesizing polymkeric substance specific target molecules to specific recognition ability.MIT is a kind of new technology grown up on the basis of the multi-door subject crossing such as chemistry, Materials science, biological chemistry.The polymkeric substance obtained through MIT is called molecularly imprinted polymer (MolecularImprintedPolymer, MIP).Compared with natural molecule recognition system, molecularly imprinted polymer not only possesses the molecule distinguishability that can compare favourably with it, also there is the high good physics and chemistry character of long service life, good stability, affinity and selectivity, thus present its advantage in fields such as antibodies mimic, bionic catalysis, biosensor, drug release, particularly have important application at separation field.
The traditional method preparing molecular engram material is entrapping method, the method embedded deeply template molecule or tension makes eluted template molecular ratio more difficult, and most of imprinted sites is embedded in polymeric inner, its adsorptive capacity is reduced, utilization ratio reduces, also can cause the destruction of molecular recognition site in process of lapping, the polymer beads obtained in addition is irregular, all poor as its current capacity and column efficiency during chromatographic stationary phases.
Summary of the invention
The present invention is to solve above-mentioned deficiency, provides a kind of preparation method and application thereof of Benzylpenicillin sodium molecular imprinted polymer on surface.
Above-mentioned purpose of the present invention is realized by following technical scheme: a kind of preparation method of Benzylpenicillin sodium molecular imprinted polymer on surface, that is: Benzylpenicillin sodium is dissolved in certain solution, described certain solvent is dimethyl sulfoxide (DMSO) (DMSO) and methyl alcohol volume ratio is the mixture of 4:1; Then, add function monomer and stir, then add linking agent, initiator, nitrogen deoxygenation, reacting by heating; Template molecule Benzylpenicillin sodium is washed away after reaction terminates, dry, obtain Benzylpenicillin sodium molecular imprinted polymer on surface.
As preferably, described function monomer is α-methacrylic acid (MAA).
As preferably, described linking agent is ethylene glycol dimethacrylate (EGDMA).
As preferably, described initiator is Diisopropyl azodicarboxylate (AIBN).
The performance study experimental technique of Benzylpenicillin sodium molecular imprinted polymer on surface of the present invention: comprise the following steps:
The preparation of a, Benzylpenicillin sodium standardized solution:
Benzylpenicillin sodium is dissolved in certain solution, is mixed with the Benzylpenicillin sodium standardized solution of different concns.
As preferably, described solvent adopts methanol solvate.
The drafting of b, typical curve:
By the Benzylpenicillin sodium solution of different concns, under 264nm wavelength, not contain the methanol solution of Benzylpenicillin sodium solution for blank, measure absorbancy respectively.Take absorbancy as ordinate zou, the concentration of penicillin solution is X-coordinate, drawing standard curve.
C, binding kinetics are tested:
Take MIP and NIP respectively, add Benzylpenicillin sodium solution, vibrate under room temperature, take out mixed solution in 24h centrifugal, get supernatant liquor ultraviolet spectrophotometer and measure its absorbancy, calculate the concentration of different adsorption time Benzylpenicillin sodium according to typical curve.Calculate polymkeric substance to the binding capacity Q (μ g/mg) of Benzylpenicillin sodium.
Q＝(C
o-C
1)·V/m
Wherein C
0, C
1represent the concentration (μ g/mL) of template molecule in solution before and after absorption respectively; V represents the volume (mL) of solution; M represents the quality (mg) of the molecularly imprinted polymer added.Draw the kinetic curve of binding capacity and binding time relation.Determine the binding capacity Q of molecularly imprinted polymer, imprinting factor α and best combination starting time.
α＝Q(MIP)/Q(NIP)
Wherein Q (MIP) represents the saturated binding capacity of molecularly imprinted polymer; Q (NIP) represents the saturated binding capacity of non-molecularly imprinted polymer.The size of imprinting factor α represents the Specific adsorption character of MIP, and its numerical value is larger, and specificity is better, and imprinting effect is stronger.
D, adsorption isothermal curve:
Take MIP, be placed in centrifuge tube, the Benzylpenicillin sodium solution of different concns.After the 20h that vibrates under room temperature reaches adsorption equilibrium, mixed solution is centrifugal, get supernatant liquor and measure absorbancy, calculate the saturated binding capacity Q of molecularly imprinted polymer, draw adsorption isothermal line.
E, specific recognition are tested:
Choose potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride as target molecule.Take MIP and NIP respectively, be placed in test tube, add Benzylpenicillin sodium solution.After vibration reaches adsorption equilibrium under room temperature, by centrifugal for each mixed solution, get supernatant liquor, measure absorbancy, the change of strength of solution before and after establishing criteria curve calculation combines.Adopt selective factor B (SelectinGFactor, β) to describe the selective recognition ability of MIP, calculate template imprinting factor-alpha
1with the imprinting factor α of template analogue
2, according to formulae discovery selective factor B β:
β＝α
1/α
2
The size of selective factor B β represents the selection separating power of MIP to template analogue, and β numerical value is larger, more easily makes template molecule and template analogue separate.
Benzylpenicillin sodium molecular imprinted polymer on surface of the present invention detects the method that the Benzylpenicillin sodium in milk in certain limit remains, and comprises the following steps:
(1) extraction of milk sample:
Get milk sample and be placed in centrifuge tube, add Extraction solvent mixing, centrifugal, repeat aforesaid operations, united extraction liquid, add another kind of extraction solution, regulate pH, heated and stirred, obtains the milk after extraction.
As preferably, described Extraction solvent is acetonitrile.
Be phosphate buffer soln (pH) as preferred described extraction solution.
(2) Method validation:
A, accuracy and withinrun precision:
Milk sample all to be assigned in 9 centrifuge tubes and to mark, respectively to the standardized solution mixing wherein adding Benzylpenicillin sodium, adding the molecularly imprinted polymer of correct amount in the solution respectively, after at room temperature concussion reaches adsorption equilibrium, by solution centrifugal.Filter, dry, weigh.According to of poor quality before and after polymkeric substance, calculate the rate of recovery and relative standard deviation RSD.
The rate of recovery (%)=measured amount/add-on × 100%
B, detectability LOD:
With the blank sample i.e. Benzylpenicillin sodium of sample not with milk, calculate standard deviation S D by the method for accuracy in Method validation and withinrun precision, then using the estimated value of 3 times of blank SD as detectability.
LOD=3S (S is the standard deviation of blank sample)
(3) mensuration of milk sample:
Add molecularly imprinted polymer MIP in centrifuge tube, add the milk sample after extraction, room temperature is shaken, centrifugal, filters, dry, weighs, and calculates the content of Benzylpenicillin sodium.
The present invention's advantage is compared with prior art: Benzylpenicillin sodium molecular imprinted polymer on surface preparation process of the present invention is simple, workable, preparation cost is cheap, fast to benzylpenicillin sodium molecular juction sum velocity, high specificity, binding capacity are high, time of equilibrium adsorption is short.Prepared Benzylpenicillin sodium molecularly imprinted polymer is high to target sample selectivity, and specificity is strong, and template molecule is recyclable the advantage such as to reuse, and can be used for the Benzylpenicillin sodium detected in milk in certain limit and remains.
Accompanying drawing explanation
Fig. 1 is the Benzylpenicillin sodium curve of adsorption kinetics figure of the molecular imprinted polymer on surface of embodiment of the present invention gained.
Fig. 2 is the Benzylpenicillin sodium adsorption isothermal curve figure of the molecular imprinted polymer on surface of embodiment of the present invention gained.
Fig. 3 is that the molecular imprinted polymer on surface of embodiment of the present invention gained is to Benzylpenicillin sodium, potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride specific recognition histogram.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
A preparation method for Benzylpenicillin sodium molecular imprinted polymer on surface, that is: be dissolved in certain solution by Benzylpenicillin sodium, and described certain solvent is dimethyl sulfoxide (DMSO) (DMSO) and methyl alcohol volume ratio is the mixture of 4:1; Then, add function monomer and stir, then add linking agent, initiator, nitrogen deoxygenation, reacting by heating; Template molecule Benzylpenicillin sodium is washed away after reaction terminates, dry, obtain Benzylpenicillin sodium molecular imprinted polymer on surface.
As preferably, described function monomer is α-methacrylic acid (MAA).
As preferably, described linking agent is ethylene glycol dimethacrylate (EGDMA).
As preferably, described initiator is Diisopropyl azodicarboxylate (AIBN).
The performance study experimental technique of Benzylpenicillin sodium molecular imprinted polymer on surface of the present invention: comprise the following steps:
The preparation of a, Benzylpenicillin sodium standardized solution:
Benzylpenicillin sodium is dissolved in certain solution, is mixed with the Benzylpenicillin sodium standardized solution of different concns.
As preferably, described solvent adopts methanol solvate.
The drafting of b, typical curve:
By the Benzylpenicillin sodium solution of different concns, under 264nm wavelength, not contain the methanol solution of Benzylpenicillin sodium solution for blank, measure absorbancy respectively.Take absorbancy as ordinate zou, the concentration of penicillin solution is X-coordinate, drawing standard curve.
C, binding kinetics are tested:
Take MIP and NIP respectively, add Benzylpenicillin sodium solution, vibrate under room temperature, take out mixed solution in 24h centrifugal, get supernatant liquor ultraviolet spectrophotometer and measure its absorbancy, calculate the concentration of different adsorption time Benzylpenicillin sodium according to typical curve.Calculate polymkeric substance to the binding capacity Q (μ g/mg) of Benzylpenicillin sodium.
Q＝(C
o-C
1)·V/m
Wherein C
0, C
1represent the concentration (μ g/mL) of template molecule in solution before and after absorption respectively; V represents the volume (mL) of solution; M represents the quality (mg) of the molecularly imprinted polymer added.Draw the kinetic curve of binding capacity and binding time relation.Determine the binding capacity Q of molecularly imprinted polymer, imprinting factor α and best combination starting time.
α＝Q(MIP)/Q(NIP)
Wherein Q (MIP) represents the saturated binding capacity of molecularly imprinted polymer; Q (NIP) represents the saturated binding capacity of non-molecularly imprinted polymer.The size of imprinting factor α represents the Specific adsorption character of MIP, and its numerical value is larger, and specificity is better, and imprinting effect is stronger.
D, adsorption isothermal curve:
Take MIP, be placed in centrifuge tube, the Benzylpenicillin sodium solution of different concns.After the 20h that vibrates under room temperature reaches adsorption equilibrium, mixed solution is centrifugal, get supernatant liquor and measure absorbancy, calculate the saturated binding capacity Q of molecularly imprinted polymer, draw adsorption isothermal line.
E, specific recognition are tested:
Choose potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride as target molecule.Take MIP and NIP respectively, be placed in test tube, add Benzylpenicillin sodium solution.After vibration reaches adsorption equilibrium under room temperature, by centrifugal for each mixed solution, get supernatant liquor, measure absorbancy, the change of strength of solution before and after establishing criteria curve calculation combines.Adopt selective factor B (SelectinGFactor, β) to describe the selective recognition ability of MIP, calculate template imprinting factor-alpha
1with the imprinting factor α of template analogue
2, according to formulae discovery selective factor B β:
β＝α
1/α
2
The size of selective factor B β represents the selection separating power of MIP to template analogue, and β numerical value is larger, more easily makes template molecule and template analogue separate.
Benzylpenicillin sodium molecular imprinted polymer on surface of the present invention detects the method that the Benzylpenicillin sodium in milk in certain limit remains, and comprises the following steps:
(1) extraction of milk sample:
Get milk sample and be placed in centrifuge tube, add Extraction solvent mixing, centrifugal, repeat aforesaid operations, united extraction liquid, add another kind of extraction solution, regulate pH, heated and stirred, obtains the milk after extraction.
As preferably, described Extraction solvent is acetonitrile.
Be phosphate buffer soln (pH) as preferred described extraction solution.
(2) Method validation:
A, accuracy and withinrun precision:
Milk sample all to be assigned in 9 centrifuge tubes and to mark, respectively to the standardized solution mixing wherein adding Benzylpenicillin sodium, adding the molecularly imprinted polymer of correct amount in the solution respectively, after at room temperature concussion reaches adsorption equilibrium, by solution centrifugal.Filter, dry, weigh.According to of poor quality before and after polymkeric substance, calculate the rate of recovery and relative standard deviation RSD.
The rate of recovery (%)=measured amount/add-on × 100%
B, detectability LOD:
With the blank sample i.e. Benzylpenicillin sodium of sample not with milk, calculate standard deviation S D by the method for accuracy in Method validation and withinrun precision, then using the estimated value of 3 times of blank SD as detectability.
LOD=3S (S is the standard deviation of blank sample)
(3) mensuration of milk sample:
Add molecularly imprinted polymer MIP in centrifuge tube, add the milk sample after extraction, room temperature is shaken, centrifugal, filters, dry, weighs, and calculates the content of Benzylpenicillin sodium.
Specific embodiment:
The present embodiment with α-methacrylic acid (MAA) for function monomer, ethylene glycol dimethacrylate (EGDMA) is linking agent, Diisopropyl azodicarboxylate (AIBN) is initiator, β-lactam antibitics Benzylpenicillin sodium is template, prepare the molecularly imprinted polymer of molecularly imprinted polymer (MIP) Benzylpenicillin sodium, detect the residual quantity of Penicillin in Milk G sodium.
As can be seen from Figure 1, along with the increase of adsorption time, the adsorptive capacity of imprinted polymer to benzylpenicillin sodium constantly rises, and substantially reaches adsorption equilibrium during 20h, and saturated extent of adsorption is 6.48 μ g/mg, imprinting factor α is 2.03.Compare with MIP, NIP is relatively weak to the absorption of Benzylpenicillin sodium.Wherein, the preparation of non-Benzylpenicillin sodium molecular imprinted polymer on surface (NIP): the preparation of non-molecularly imprinted polymer (NIP) is not except adding template molecule Benzylpenicillin sodium, and preparation method is identical with MIP.
Experiment shows, imprinted polymer MIP surface is containing the binding site matched with template molecule Benzylpenicillin sodium with in conjunction with hole.The Benzylpenicillin sodium trace hole on imprinted polymer surface and duct are very shallow, and Benzylpenicillin sodium molecular diffusion resistance is little, are easy to enter trace hole and contact with binding site.
As can be seen from Figure 2, along with the increase of concentration, the saturated binding capacity Q of molecularly imprinted polymer is in the trend increased, and illustrating that the binding site of MIP to Benzylpenicillin sodium is of equal value, is all noncovalent interaction power.Maximum apparent binding capacity is 6.48 μ g/mg.
Equilibrium adsorption is adopted to determine imprinted polymer to Benzylpenicillin sodium, potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride specific recognition adsorptive capacity.As can be seen from Figure 3, imprinted polymer is obviously better than potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride to the adsorptive power of Benzylpenicillin sodium.The trace hole of a large amount of Benzylpenicillin sodium is distributed with in the polymer foil on imprinted polymer surface, these holes in size, space structure and action site etc. with Benzylpenicillin sodium molecule be matched, low with the matching degree of potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride, cause them to be obviously weaker than Benzylpenicillin sodium by adsorptive power.Above-mentioned experimental result indicates imprinted polymer has special identification selection and height binding affinity to Benzylpenicillin sodium.The adsorptive capacity of MIP to different target molecules has larger difference, shows selective adsorption performance.But the adsorptive capacity of NIP to different target molecule does not have obvious difference.This is because MIP inside exists and template molecule Benzylpenicillin sodium complementary molecule trace hole in shape, and interactional binding site can be formed with Benzylpenicillin sodium molecule, under the acting in conjunction of molecular imprinting hole and binding site, MIP just can optionally identify Benzylpenicillin sodium molecule.
From experimental result, the rate of recovery of Benzylpenicillin sodium is 89.23%, has the good rate of recovery, and relative standard deviation is 1.21%.Experimental result shows: the accuracy of the method is higher.Benzylpenicillin sodium imprinted polymer has stronger adsorptive power to the Benzylpenicillin sodium in milk; Detectability LOD=3S=3 × 1.01=3.03mg.Experimental result shows: LOD=3.03mg, illustrates that the method has certain sensitivity.Benzylpenicillin sodium molecularly imprinted polymer has stronger adsorptive power to standard penicillium G sodium solution.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize specification sheets of the present invention and embodiment content to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical fields, be all in like manner included in scope of patent protection of the present invention.
Claims (7)
1. a preparation method for Benzylpenicillin sodium molecular imprinted polymer on surface, is characterized in that: Benzylpenicillin sodium is dissolved in certain solution, and described certain solvent is dimethyl sulfoxide (DMSO) (DMSO) and methyl alcohol volume ratio is the mixture of 4:1; Then, add function monomer and stir, then add linking agent, initiator, nitrogen deoxygenation, reacting by heating; Template molecule Benzylpenicillin sodium is washed away after reaction terminates, dry, obtain Benzylpenicillin sodium molecular imprinted polymer on surface.
2. the preparation method of a kind of Benzylpenicillin sodium molecular imprinted polymer on surface according to claim 1, is characterized in that: described function monomer is α-methacrylic acid (MAA).
3. the preparation method of a kind of Benzylpenicillin sodium molecular imprinted polymer on surface according to claim 1, is characterized in that: described linking agent is ethylene glycol dimethacrylate (EGDMA).
4. the preparation method of a kind of Benzylpenicillin sodium molecular imprinted polymer on surface according to claim 1, is characterized in that: described initiator is Diisopropyl azodicarboxylate (AIBN).
5. the preparation method of a kind of Benzylpenicillin sodium molecular imprinted polymer on surface according to claim 1, is characterized in that: the Benzylpenicillin sodium that described Benzylpenicillin sodium molecular imprinted polymer on surface can detect in milk in certain limit remains.
6. the performance study experimental technique of Benzylpenicillin sodium molecular imprinted polymer on surface: it is characterized in that: comprise the following steps:
The preparation of a, Benzylpenicillin sodium standardized solution:
Benzylpenicillin sodium is dissolved in certain solution, is mixed with the Benzylpenicillin sodium standardized solution of different concns.
As preferably, described solvent adopts methanol solvate.
The drafting of b, typical curve:
By the Benzylpenicillin sodium solution of different concns, under 264nm wavelength, not contain the methanol solution of Benzylpenicillin sodium solution for blank, measure absorbancy respectively.Take absorbancy as ordinate zou, the concentration of penicillin solution is X-coordinate, drawing standard curve.
C, binding kinetics are tested:
Take MIP and NIP respectively, add Benzylpenicillin sodium solution, vibrate under room temperature, take out mixed solution in 24h centrifugal, get supernatant liquor ultraviolet spectrophotometer and measure its absorbancy, calculate the concentration of different adsorption time Benzylpenicillin sodium according to typical curve.Calculate polymkeric substance to the binding capacity Q (μ g/mg) of Benzylpenicillin sodium.
Q＝(C
o-C
1)·V/m
Wherein C
0, C
1represent the concentration (μ g/mL) of template molecule in solution before and after absorption respectively; V represents the volume (mL) of solution; M represents the quality (mg) of the molecularly imprinted polymer added.Draw the kinetic curve of binding capacity and binding time relation.Determine the binding capacity Q of molecularly imprinted polymer, imprinting factor α and best combination starting time.
α＝Q(MIP)/Q(NIP)
Wherein Q (MIP) represents the saturated binding capacity of molecularly imprinted polymer; Q (NIP) represents the saturated binding capacity of non-molecularly imprinted polymer.The size of imprinting factor α represents the Specific adsorption character of MIP, and its numerical value is larger, and specificity is better, and imprinting effect is stronger.
D, adsorption isothermal curve:
Take MIP, be placed in centrifuge tube, the Benzylpenicillin sodium solution of different concns.After the 20h that vibrates under room temperature reaches adsorption equilibrium, mixed solution is centrifugal, get supernatant liquor and measure absorbancy, calculate the saturated binding capacity Q of molecularly imprinted polymer, draw adsorption isothermal line.
E, specific recognition are tested:
Choose potassium v calcium, amoxycilline Trihydrate bp, Cefdinir, Cefixime Micronized, Cephradine, levofloxacin hydrochloride as target molecule.Take MIP and NIP respectively, be placed in test tube, add Benzylpenicillin sodium solution.After vibration reaches adsorption equilibrium under room temperature, by centrifugal for each mixed solution, get supernatant liquor, measure absorbancy, the change of strength of solution before and after establishing criteria curve calculation combines.Adopt selective factor B (SelectinGFactor, β) to describe the selective recognition ability of MIP, calculate template imprinting factor-alpha
1with the imprinting factor α of template analogue
2, according to formulae discovery selective factor B β:
β＝α
1/α
2
The size of selective factor B β represents the selection separating power of MIP to template analogue, and β numerical value is larger, more easily makes template molecule and template analogue separate.
7. Benzylpenicillin sodium molecular imprinted polymer on surface detects the method that the Benzylpenicillin sodium in milk in certain limit remains, and it is characterized in that: comprise the following steps:
(1) extraction of milk sample:
Get milk sample and be placed in centrifuge tube, add Extraction solvent mixing, centrifugal, repeat aforesaid operations, united extraction liquid, add another kind of extraction solution, regulate pH, heated and stirred, obtains the milk after extraction.
As preferably, described Extraction solvent is acetonitrile.
Be phosphate buffer soln (pH) as preferred described extraction solution.
(2) Method validation:
A, accuracy and withinrun precision:
Milk sample all to be assigned in 9 centrifuge tubes and to mark, respectively to the standardized solution mixing wherein adding Benzylpenicillin sodium, adding the molecularly imprinted polymer of correct amount in the solution respectively, after at room temperature concussion reaches adsorption equilibrium, by solution centrifugal.Filter, dry, weigh.According to of poor quality before and after polymkeric substance, calculate the rate of recovery and relative standard deviation RSD.
The rate of recovery (%)=measured amount/add-on × 100%
B, detectability LOD:
With the blank sample i.e. Benzylpenicillin sodium of sample not with milk, calculate standard deviation S D by the method for accuracy in Method validation and withinrun precision, then using the estimated value of 3 times of blank SD as detectability.
LOD=3S (S is the standard deviation of blank sample)
(3) mensuration of milk sample:
Add molecularly imprinted polymer MIP in centrifuge tube, add the milk sample after extraction, room temperature is shaken, centrifugal, filters, dry, weighs, and calculates the content of Benzylpenicillin sodium.
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|---|---|---|---|
| CN201510934634.9A CN105384872A (en) | 2015-12-15 | 2015-12-15 | Preparation method and use of penicillin G sodium salt surface molecular imprinting polymer |
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