Polyacrylamide gel beads for the recognition of staphylococcal enterotoxin B

Protein‐imprinted polyacrylamide gel beads (IPGB) were synthesized via inverse suspension polymerization, using staphylococcal enterotoxin B (SEB) as template. The adsorption capacity of SEB‐IPGB was almost three times as much as that of non‐imprinted gel beads. The Langmuir adsorption models were applied to describe the equilibrium isotherms. The results showed that an equal class of adsorption was formed in the SEB‐IPGB with the maximum adsorption capacity of 8.40 mg SEB/g imprinted beads. The selectivity test of imprinted beads shows that they exhibited good recognition for SEB as compared with the other proteins. The formation of multiple hydrogen bonds and complementary shape between the imprinting cavities and the template proteins would be the two factors that led to the imprinting effect. The obtained SEB‐IPGB would be used as a potential material for protein toxin separation, extraction, and purification. Copyright © 2014 John Wiley & Sons, Ltd.     

光接枝表面修饰法制备牛血红蛋白的分子印迹微球

聚苯乙烯球载体表面经引发转移终止剂修饰后, 采用光接枝表面印迹方法制备了以牛血红蛋白(BHb)为模板分子、丙烯酰胺为功能单体和N,N′-亚甲基双丙烯酰胺为交联剂的分子印迹聚合物微球(MIP). 进一步采用红外光谱(IR)、扫描电子显微镜(SEM)和元素分析对聚合物微球进行了表征, 证实了载体表面成功地接枝了分子印迹层, 并研究了其吸附性能和分子识别选择性能. 结果表明, 采用光接枝表面修饰法制备的分子印迹微球对模板分子有着很好的吸附容量和识别选择性.     

Preparation of l‐phenylalanine‐imprinted solid‐phase extraction sorbent by Pickering emulsion polymerization and the selective enrichment of l‐phenylalanine from human urine

A novel l‐ phenylalanine molecularly imprinted solid‐phase extraction sorbent was synthesized by the combination of Pickering emulsion polymerization and ion‐pair dummy template imprinting. Compared to other polymerization methods, the molecularly imprinted polymers thus prepared exhibit a high specific surface, large pore diameter, and appropriate particle size. The key parameters for solid‐phase extraction were optimized, and the result indicated that the molecularly imprinted polymer thus prepared exhibits a good recovery of 98.9% for l‐ phenylalanine. Under the optimized conditions of the procedure, an analytical method for l‐ phenylalanine was well established. By comparing the performance of the molecularly imprinted polymer and a commercial reverse‐phase silica gel, the obtained molecularly imprinted polymer as an solid‐phase extraction sorbent is more suitable, exhibiting high precision (relative standard deviation 3.2%, n = 4) and a low limit of detection (60.0 ± 1.9 nmol·L^?1) for the isolation of l‐ phenylalanine. Based on these results, the combination of the Pickering emulsion polymerization and ion‐pair dummy template imprinting is effective for preparing selective solid‐phase extraction sorbents for the separation of amino acids and organic acids from complex biological samples.     

Preconcentration of indapamide from human urine using molecularly imprinted solid‐phase extraction

A simple, sensitive, and selective molecularly imprinted solid‐phase extraction and spectrophotometric method has been developed for the clean‐up and preconcentration of indapamide from human urine. Molecularly imprinted polymers were prepared by a non‐covalent imprinting approach using indapamide as a template molecule, 2‐(trifluoromethyl) acrylic acid as a functional monomer, ethylene glycol dimethacrylate as a crosslinker, N,N‐azobisisobutyronitrile as a thermal initiator and acetonitrile as a porogenic solvent. A non‐imprinted polymer was also prepared in the same way, but in the absence of template. Molecularly imprinted polymer and non‐imprinted polymer sorbents were dry‐packed into solid‐phase extraction cartridges. Eluates from cartridges were analyzed using a spectrophotometer for the determination of indapamide by referring to the calibration curve in the range 0.14–1.50 μg/mL. Preconcentration factor, limit of detection, and limit of quantification were 16.30, 0.025 μg/mL, and 0.075 μg/mL, respectively. A relatively high imprinting factor (9.3) was also achieved and recovery values for the indapamide spiked into human urine were in the range of 80.1–81.2%. In addition, relatively low within‐day (0.17–0.42%) and between‐day (1.1–1.4%) precision values were obtained as well. The proposed molecularly imprinted solid‐phase extraction and spectrophotometric method was successfully applied to selective extraction, preconcentration, and determination of indapamide from human urine samples.     

Cost‐effective imprinting combining macromolecular crowding and a dummy template for the fast purification of punicalagin from pomegranate husk extract

The combination of molecular crowding and virtual imprinting was employed to develop a cost‐effective method to prepare molecularly imprinted polymers. By using linear polymer polystyrene as a macromolecular crowding agent, an imprinted polymer recognizable to punicalagin had been successfully synthesized with punicalin as the dummy template. The resulting punicalin‐imprinted polymer presented a remarkable selectivity to punicalagin with an imprinting factor of 3.17 even at extremely low consumption of the template (template/monomer ratio of 1:782). In contrast, the imprinted polymer synthesized without crowding agent, did not show any imprinting effect at so low template amount. The imprinted polymers made by combination of molecular crowding and virtual imprinting can be utilized for the fast separation of punicalagin from pomegranate husk extract after optimizing the protocol of solid‐phase extraction with the recovery of 85.3 ± 1.2%.     

Mimicking a Receptor for Cyanide Ion Based on Ion Imprinting and Its Applications in Potential Transduction

Using the strategy of template polymerization, a presynthesized specific metal‐complexing polymer (poly(methacryloylhistidine‐Ni(II)‐CN^?), Ni‐CN/IP) has been specifically used to recognize cyanide ion. As described previously, nickel(II)‐methacryloylhistidine dihydrate complex monomer was synthesized and reacted with KCN to produce the monomer‐template complex. This monomer‐template complex phase was polymerized in a dispersion medium. After polymerization, the template (CN^?) was removed from the Ni‐CN/IP, producing CN^? ion imprinted metal‐chelate polymer. The synthesized ion imprinted polymer is examined as a novel potential cyanide selective ionophore in polymeric membrane type ion selective electrodes. Membranes formulated with Ni‐CN/IP are shown to exhibit enhanced potentiometric selectivity for cyanide over more lipophilic anions including perchlorate, iodide, and thiocyanate. Addition of lipophilic cationic sites into the organic membranes enhanced the response and selectivity towards CN^? ion, while addition of lipophilic anionic sites deteriorated the response but enhanced the selectivity, indicating that the Ni‐CN/IP particles behaves via the so‐called “mixed‐mode” response mechanism. The fabricated sensors possessed good performance characteristics, in terms of life span, selectivity for CN^? ion over a wide range of other interfering anions, fast response, stability and high reproducibility. Applications for direct determination of cyanide ion in hazardous wastes using the proposed sensors showed good correlation with data obtained using commercial solid state cyanide electrode, with no significant difference in the t‐test values with 95 % confidence level. An F‐test revealed that the standard deviations of the replicate sample measurements obtained by the two methods were not significantly different.     

Isolation of sinapic acid from broccoli using molecularly imprinted polymers

A molecularly imprinted polymer was synthesized for the purpose of sinapic acid isolation from Egyptian nutraceutical Botrytis italica, L. (broccoli) due to its prominent medicinal and wide pharmacological activities. A computational study was first developed to determine the optimal template to functional monomer molar ratio. Based on the computational results, five polymers were synthesized using a bulk polymerization method with sinapic acid as the template molecule. Evaluation of the synthesized polymers binding performance was carried out using batch rebinding assay, which revealed that the molecularly imprinted polymer of molar ratio (1:4:20), template to functional monomer (4‐vinyl pyridine) to crosslinker (ethylene glycol dimethacrylate) was of optimum performance, thus, this polymer was applied for sinapic acid isolation from closely related analogues. This represents a more practical approach to isolate sinapic acid from different natural extracts selectively.     

Advances in the development of molecularly imprinted polymers for the separation and analysis of proteins with liquid chromatography

This review documents recent advances in the design, synthesis, characterization, and application of molecularly imprinted polymers in the form of monoliths and particles/beads for the use in the separation and analysis of proteins with solid‐phase extraction or liquid chromatography. The merits of three‐dimensional molecular imprinting, whereby the molecular template is randomly embedded in the polymer, and two‐dimensional imprinting, in which the template is confined to the surface, are described. Target protein binding can be achieved by either using the entire protein as a template or by using a protein substructure as template, that is, a peptide, as in the "epitope" approach. The intended approach and strategy then determine the choice of polymerization method. A synopsis has been provided on methods used for the physical, chemical, and functional characterizations and associated performance evaluations of molecularly imprinted and nonimprinted control polymers, involving a diverse range of analytical techniques commonly used for low and high molecular mass analytes. Examples of recent applications demonstrate that, due to the versatility of imprinting methods, molecularly imprinted monoliths or particles/beads can be adapted to protein extraction/depletion and separation procedures relevant to, for example, protein biomarker detection and quantification in biomedical diagnostics and targeted proteomics.     

球形钴离子模板缩聚物的制备及其吸附性能研究

以钴离子为模板,四乙烯五胺为功能基配体,反相悬浮聚合制备出球形钴离子模板缩聚物。该模板缩聚物对钴离子的吸附性能显著优于相应的非模板缩聚物。说明应用离子模板技术可提高普通吸附树脂的吸附性能。可望在环境、医学等领域获得实际应用。     

Preparation and application of molecularly imprinted polymer for isolation of chicoric acid from Chicorium intybus L. medicinal plant

Molecularly imprinted polymer (MIP) was synthesized and applied for the extraction of chicoric acid from Chicory herb (Chicorium intybus L.). A computational study was developed to find a suitable template to functional monomer molar ratio for MIP preparations. The molar ratio was chosen based on the comparison of the binding energy of the complexes between the template and functional monomers. Based on the computational results, eight different polymers were prepared using chicoric acid as the template. The MIPs were synthesized in a non-covalent approach via thermal free-radical polymerization, using two different polymerization methods, bulk and suspension. Batch rebinding experiments were performed to evaluate the binding properties of the imprinted polymers. The best results were obtained with a MIP prepared using bulk polymerization with 4-vinylpyridine (4-VP) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the crosslinker with a molar ratio of 1:4:20. The best MIP showed selective binding ability toward chicoric acid in the presence of the template’s structural analogues, caffeic acid, caftaric acid and chlorogenic acid.     

Preparation of bovine hemoglobin-imprinted polymer beads via the photografting surface-modified method

Molecularly imprinted polymers (MIPs), based on photografting surface-modified polystyrene beads as matrices, were prepared with acrylamide as the functional monomer, bovine hemoglobin as the template molecule and N, N′-methylene bisacrylamide as the crosslinker in a phosphate buffer. The results of IR, scanning electron microscope (SEM) and elemental analyses demonstrated the formation of a grafting polymer layer on the polystyrene-bead surface. Subsequent removal of the template left behind cavities on the surface of the polymer matrix with a shape and an arrangement of functional groups having complementary binding sites with the original template molecule. The adsorption studies showed that the imprinted polymers have a good adsorption capacity and specific recognition for bovine hemoglobin as the template molecule. Our results demonstrated that the polymer prepared via the photografting surface-modified method exhibited better selectivity for the template. Attempts to employ the new method in molecular imprinting techniques may introduce new applications for MIPs and facilitate probable protein separation and purification. __________ Translated from Chemical Journal of Chinese Universities, 2008, 29(1): 64–70     

One‐step preparation of magnetic imprinted nanoparticles adopting dopamine‐cupric ion as a co‐monomer for the specific recognition of bovine hemoglobin

A novel magnetic core–shell polydopamine–cupric ion complex imprinted polymer was prepared in one‐step through surface imprinting technology, which could specifically recognize bovine hemoglobin from the real blood samples. The polymerization conditions and adsorption performance of the resultant nanomaterials were investigated in detail. The results showed that the cupric ion played an important role in the recognition of template proteins. The saturating adsorption capacity of this kind of imprinted polymers was 2.23 times greater than those of imprinted polymers without cupric ion. The imprinting factor of the imprinted materials was as high as 4.23 for the template molecule. The selective separation bovine hemoglobin from the real blood sample is successfully applied. In addition, the prepared materials had excellent stability and no obvious deterioration after five adsorption–regeneration cycles. Easy preparation, rapid separation, high binding capacity and satisfactory selectivity for the template protein make this polymer attractive in the separation of high‐abundance proteins.     

咪草烟分子印迹聚合物的制备及其选择性吸附性能的研究

以丙烯酸为聚合单体,TMPTA为交联剂,AIBN为引发剂,咪草烟为模板分子,采用在低温光聚合的方法,制备了对咪草烟分子具有选择性识别能力的分子印迹聚合物．通过IR和HPLC表征,咪草烟分子印迹聚合物对咪草烟分子具有良好的识别作用．     

Recognition of Staphylococcus enterotoxin via molecularly imprinted beads

The synthesis of molecularly imprinted beads for the recognition of the protein Staphylococcus enterotoxin B (SEB) is described. Two kinds of organic silane (3-aminopropyltrimethoxysilane (APTMS) and octyltrimethoxysilane (OTMS)) were polymerized on the surface of polystyrene microspheres after the SEB template was covalently immobilized by forming imine bonds. The resulting imprinted beads were selective for SEB. The Langmuir adsorption models were applied to describe the equilibrium isotherms. The results showed that an equal class of adsorption was formed in the molecularly imprinted polymer (MIP) with the maximum adsorption capacity of 3.86 mg SEB/g imprinted beads. The MIP has much higher adsorption capacity for SEB than the nonimprinted polymer, and the MIP beads have a higher selectivity for the template molecule.     

A novel method to prepare monolithic molecular imprinted polymer fiber for solid‐phase microextraction by microwave irradiation†

In this paper, a new approach to prepare monolithic molecularly imprinted polymer (MIP) fibers for solid‐phase microextraction is proposed with the help of microwave irradiation. Imprinting polymerization was carried out within silica capillaries in 4.5 min, using dimethyl phthalate (DMP) as a template molecular, α‐methacrylic acid as a functional monomer and ethylene dimethacrylate as a crosslinker, acetonitrile as the porogenic solvent. The synthesis was optimized by varying the ratio of template/monomer and different volume of porogen. The resulted MIP fibers were obtained after silica being etched away with a controlled length of 1 cm, and subsequently characterized by SEM. In order to increase the selective extraction of DMP, factors affecting the extraction including extraction time, salt concentration, desorption time, and desorption solvents were investigated for solid‐phase microextraction procedures in detail. The selectivity coefficients, defined as the extraction amount ratio of MIP to its nonimprinting fiber, were 5.6, 2.6, and 1.4 for DMP and its counterpart including dibutyl phthalate and di‐n‐octylo‐phthalate, respectively. The resulted fibers were also applied to detect DMP, dibutyl phthalate, and di‐n‐octylo‐phthalate in bottled beverage samples coupled to HPLC and resulted in relative recoveries of up to 73.8–98.5%, respectively.     

Synthesis of novel ion‐imprinted polymeric nanoparticles based on dibenzo‐21‐crown‐7 for the selective pre‐concentration and recognition of rubidium ions

In this work, we report the first application of ion‐imprinted technology via precipitation polymerization for simple and practical determination of rubidium ions. The rubidium‐ion‐imprinted polymer nanoparticles were prepared using dibenzo‐21‐crown‐7 as a selective ligand, methacrylic acid as functional monomer, ethylene glycol dimethacrylate as cross linker, and 2,2′‐azobisisobutyronitrile as radical initiator. The resulting powder material was characterized using scanning electron microscopy, which showed colloidal nanoparticles of 100–200 nm in diameter and slightly irregular in shape. The maximum adsorption capacity of the ion imprinted particles was 63.36 μmol/g. The experimental conditions such as nature and concentration of eluent, pH, adsorption and desorption times, weight of the polymer material, aqueous phase and desorption agent volumes were also studied. Finally, selectivity of the prepared IIP particles toward rubidium ion was investigated in the presence of some foreign metal ions.     

分子印迹聚合物的制备及其对香草醛的吸附行为

以香草醛为模板分子、甲基丙烯酸为功能单体、乙二醇二甲基丙烯酸酯为交联剂,制备了香草醛分子印迹聚合物(MIP);研究了以不同致孔剂合成的MIP在水溶液中对香草醛的吸附行为.结果表明,以乙腈为致孔剂合成的MIP对香草醛具有较高的识别特性,能较简便地用于香草醛的分离和富集.     

Novel metal‐ion‐mediated,complex‐imprinted solid‐phase microextraction fiber for the selective recognition of thiabendazole in citrus and soil samples

A novel metal‐ion‐mediated complex‐imprinted‐polymer‐coated solid‐phase microextraction (SPME) fiber used to specifically recognize thiabendazole (TBZ) in citrus and soil samples was developed. The complex‐imprinted polymer was introduced as a novel SPME coating using a “complex template” constructed with Cu(II) ions and TBZ. The recognition and enrichment properties of the coating in water were significantly improved based on the metal ion coordination interaction rather than relying on hydrogen bonding interactions that are commonly applied for the molecularly imprinting technique. Several parameters controlling the extraction performance of the complex‐imprinted‐polymer‐coated fiber were investigated including extraction solvent, pH value, extraction time, metal ion species, etc. Furthermore, SPME coupled with HPLC was developed for detection of TBZ, and the methods resulted in good linearity in the range of 10.0–150.0 ng/mL with a detection limit of 2.4 ng/mL. The proposed method was applied to the analysis of TBZ in spiked soil, orange, and lemon with recoveries of 80.0–86.9% and RSDs of 2.0–8.1%. This research provides an example to prepare a desirable water‐compatible and specifically selective SPME coating to extract target molecules from aqueous samples by introducing metal ions as the mediator.     

葡萄球菌肠毒素B分子印迹聚合物的制备及其与蛋白质的结合性能

以葡萄球菌肠毒素B(SEB)蛋白为模板分子,以聚苯乙烯微球为基质,采用表面分子印迹法制备了SEB分子印迹聚合物.利用平衡吸附试验分析了SEB聚合物对目标蛋白的吸附能力及对类似底物的选择性;分析了该聚合物的吸附动力学,并利用扫描电镜观察了其形貌特征和颗粒尺寸.结果表明,经Scatchard模型分析求得的标题聚合物的最大表观结合量Qmax为3.23mg/g;所制备的SEB分子印迹聚合物呈微球形,粒径约为1²μm,对SEB蛋白具有较好的吸附性和特异选择性.     

Preparation and selective recognition of a novel solid‐phase microextraction fiber combined with molecularly imprinted polymers for the extraction of parabens in soy sample

A prepared molecularly imprinted polymer with ethyl p‐hydroxybenzoate as template molecule was applied for the first time to a homemade solid‐phase microextraction fiber. The molecularly imprinted polymer‐coated solid‐phase microextraction fiber was characterized by scanning electron microscopy and thermogravimetric analysis. Various parameters were investigated, including extraction temperature, extraction time, and desorption time. Under the optimum extraction conditions, the molecularly imprinted polymer‐coated solid‐phase microextraction fiber exhibited higher selectivity with greater extraction capacity toward parabens compared with the nonimprinted polymer‐coated solid‐phase microextraction fiber and commercial fibers. The molecularly imprinted polymer‐coated solid‐phase microextraction fiber was tested using gas chromatography to determine parabens, including methyl p‐hydroxybenzoate, ethyl p‐hydroxybenzoate, and propyl p‐hydroxybenzoate. The linear ranges were 0.01–10 μg/mL with a correlation coefficient above 0.9943. The detection limits (under signal‐to‐noise ratio of 3) were below 0.30 μg/L. The fiber was successfully applied to the simultaneous analysis of three parabens in spiked soy samples with satisfactory recoveries of 95.48, 97.86, and 92.17%, respectively. The relative standard deviations (n=6) were within 2.83–3.91%. The proposed molecularly imprinted polymer‐coated solid‐phase microextraction method is suitable for selective extraction and determination of trace parabens in food samples.