Metal ion-selective membrane prepared by surface molecular imprinting

Surface molecular imprinting was applied to the preparation of an ion-selective polymeric membrane for the first time. The use of acrylonitrile-butadiene rubber and a porous solid support in the polymer matrix resulted in improved flexibility and mechanical strength of the imprinted membrane. The asymmetric porous structure of the membrane was observed by scanning electron microscopy. The selectivity of the zinc(II)-imprinted membrane was evaluated by competitive adsorption and permeation studies. The imprinted membrane showed higher adsorption affinity and permeation selectivity towards the imprinted zinc ion than the non-imprinted counterpart. On the basis of the results obtained, the permeation mechanism of the metal ions was considered to be hopping of metal ions on the binding sites in the membranes.     

Synthesis and evaluation of uniformly sized nalidixic acid-imprinted nanospheres based on precipitation polymerization method for analytical and biomedical applications

For the first time in this work, uniform molecularly imprinted polymer (MIP) nanoparticles were prepared using nalidixic acid as a template. The MIP nanoparticles were successfully synthesized by precipitation polymerization applying methacrylic acid (MAA) as a functional monomer and trimethylolpropane trimethacrylate (TRIM) as a cross-linking monomer at different mole ratios. The morphology, binding, recognition, selectivity, and in vitro release behaviors of obtained particles were studied. The produced polymers were characterized by Fourier transform infrared spectroscopy and differential scanning calorimetric. Furthermore, their morphology was analyzed accurately by scanning electron microscopy, photon correlation spectroscopy, and Brunauer-Emmett-Teller analysis. The nanospheres and microspheres with mean diameter values of 94 nm, 256 nm, and 1.2 μm were obtained using nalidixic acid-MAA-TRIM various mole ratios. Among the MIPs, the product with nalidixic acid-MAA-TRIM mole ratio of 1:12:12 established nanospheres with the lowest polydispersity index (0.003), an average pore diameter (12 nm), and the highest specific surface area (280 m(2) g(-1)) and selectivity factor (10.4). Results from binding experiments demonstrated that the imprinted nanospheres with a 94-nm mean diameter and a binding capacity of 28 mg of nalidixic acid per gram of polymer had higher specific affinity to nalidixic acid in contrast with the other imprinted nanospheres, microspheres, and nonimprinted particles. However, the binding performance of imprinted nanospheres in human serum was estimated using high-performance liquid chromatography analysis (binding approximately 98% of nalidixic acid). In addition, release experiments proved to be successful in the controlled release of nalidixic acid during a long period. The 20% of loaded nalidixic acid was released from the imprinted nanospheres within the first 20 h, whereas the remaining 80% was released in the after 120 h. The nalidixic acid release kinetics from the MIPs was highly affected by properties of the particles.     

In situ synthesis of molecularly imprinted nanoparticles in porous support membranes using high-viscosity polymerization solvents

There is a growing need in membrane separations for novel membrane materials providing selective retention. Molecularly imprinted polymers (MIPs) are promising candidates for membrane functionalization. In this work, a novel approach is described to prepare composite membrane adsorbers incorporating molecularly imprinted microparticles or nanoparticles into commercially available macroporous filtration membranes. The polymerization is carried out in highly viscous polymerization solvents, and the particles are formed in situ in the pores of the support membrane. MIP particle composite membranes selective for terbutylazine were prepared and characterized by scanning electron microscopy and N? porosimetry. By varying the polymerization solvent microparticles or nanoparticles with diameters ranging from several hundred nanometers to 1 μm could be embedded into the support. The permeability of the membranes was in the range of 1000 to 20,000 Lm?2 hr?1 bar?1. The imprinted composite membranes showed high MIP/NIP (nonimprinted polymer) selectivity for the template in organic media both in equilibrium-rebinding measurements and in filtration experiments. The solid phase extraction of a mixture of the template, its analogs, and a nonrelated compound demonstrated MIP/NIP selectivity and substance selectivity of the new molecularly imprinted membrane. The synthesis technique offers a potential for the cost-effective production of selective membrane adsorbers with high capacity and high throughput.     

Stereoselective histidine sensor based on molecularly imprinted sol-gel films

A piezoelectric sensor coated with a thin molecularly imprinted sol-gel film has been developed for the determination of L-histidine in the liquid phase. Without preprotection, L-histidine was imprinted directly into silica sol-gel films that consisted of a hybrid mixture of functionalized organosilicon precursors (phenyltrimethoxysilane and methyltrimethoxysolane). The viscoelasticity of the film in the air and in buffer solution has been studied by the piezoelectric quartz crystal impedance technique. The binding of L-histidine to the imprinted film in the liquid phase was investigated by the piezoelectric microgravimetry and electrochemical impedance technique. Scatchard analysis showed that the maximum binding site (Qmax) of the L-histidine imprinted sol-gel film is about 23.7 micromol/g. A linear range from 5.0x10(-8) to 1.0x10(-4) M for a detection of L-histidine has been observed with a detection limit of 2.5x10(-8) M for S/N=3. The proposed imprinted sol-gel sensor exhibits good stability, high specificity, and excellent stereoselectivity.     

Design and evaluation of thin and flexible theophylline imprinted polymer membrane materials

The aim of this work was to produce a thin, flexible and diffusion able molecularly imprinted polymeric matrix with good template accessibility. Membranes were prepared using a non‐covalent molecular imprinting approach and their physical characteristics and binding capabilities investigated. Two materials were used, a poly(tri‐ethyleneglycol dimethyacrylate‐co‐methyl methacrylate‐co‐methacrylic acid) copolymer containing 14% cross‐linker and a monomer (g) to porogen (ml) ratio of 1:0.5 (A), and a blend of poly(TEGMA‐co‐MAA) and polyurethane (B). The polyurethane was added to improve membrane flexiblity and stability. The polymers were characterized using AFM, SEM and nitrogen adsorption, whilst binding was evaluated using batch‐rebinding studies. For all membranes the specific surface area was low (<10 m²/g). MIP (A) films were shown to bind specifically at low concentrations but specific binding was masked by non‐specific interactions at elevated concentrations. Selectivity studies confirmed specificity at low concentrations. K_D approximations confirmed a difference in the population of binding sites within NIP and MIP films. The data also indicated that at low concentrations the ligand‐occupied binding site population approached homogeneity. Scanning electron microscopy images of membrane (B) revealed a complex multi‐layered system, however these membranes did not demonstrate specificity for the template. The results described here demonstrate how the fundamental parameters of a non‐covalent molecularly imprinted system can be successfully modified in order to generate flexible and physically tolerant molecularly imprinted thin films. Copyright © 2009 John Wiley & Sons, Ltd.     

Composite matrix membranes as synthetic receptor systems. 2. Structural-morphological characteristics

Structural and morphological characteristics of composite imprinted membranes for selective recognizing of adenosine 3',5'-cyclic monophosphate (cAMP) were studied. Composite polyvinylidene fluoride microfiltration membranes (Millipore) covered with a thin imprinted polymer layer were prepared using photoinitiated copolymerization of dimethylaminoethylmetacrylate with trimethylolpropanethrimethacrylate in the presence of cAMP as template. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) were used to visualise surfaces and cross-sections of imprinted membranes and to determine their structural and morphological parameters such as pore size, thickness of selective imprinted layer, surface roughness as well as total surface contact area. The impact of structural characteristics on separation properties of the imprinted membranes was analyzed. It was found out that the thickness of the imprinted polymer layer with optimal recognizing properties is limited.     

Supported imprinted nanospheres for the selective recognition of cholesterol

The preparation of innovative polymeric systems using molecular imprinting technology for application in extracorporeal blood purification is described. Membranes based on a methylmethacrylate-co-acrylic acid copolymer, produced through the phase inversion method, were modified introducing into their structure specific binding sites for cholesterol molecule by adding molecularly imprinted nanoparticles in the membrane matrix. Membranes prepared are intended to selectively remove cholesterol from the blood by using interactions at a molecular level, between the membrane/nanoparticles devices and the template, created during the preparation of polymers. Three polymeric systems in form of nanoparticles were prepared differing in the polymerisation solvent (a mixture of acetonitrile and ethanol (1:1) or pure ethanol), and the molar ratio between the functional monomer and the cross-linker (2.3:1 and 1:1). Two out of three of the prepared polymers showed a very good template rebinding capacity both in phosphate buffer solution (pH 6.9) and in ethanol. In particular the nanoparticles rebound 115.4 mg cholesterol/g polymer in buffer solution, and 57 mg cholesterol/g polymer in ethanol.

The deposition of the nanoparticles on the surface of the phase inversion membranes produced devices with interesting rebinding performances towards cholesterol in buffer solution: a specific recognition of 14.09 mg cholesterol/g system (membrane and nanoparticles) was detected, indicating maintained binding capacity of supported particles as well.     

Molecularly imprinted polymer of 5-methyluridine for solid-phase extraction of pyrimidine nucleoside cancer markers in urine

Normal and modified urinary nucleosides represent potential biomarkers for cancer diagnosis. To selectively extract modified nucleosides, we developed a molecularly imprinted polymer (MIP) of 5-methyluridine as selective material for molecularly imprinted solid-phase extraction (MISPE). The MIPs were obtained from vinyl-phenylboronate ester derivative of the template, acrylamide and pentaerythritol triacrylate co-polymer, and were tested in batch and cartridge experiments with aqueous samples. Our results indicated that the imprinted polymer was selective for pyrimidine nucleosides with a K(d) and a B(max) of 46 microM and 18 micromol/g, respectively. Finally, a MISPE of the most common pyrimidine nucleoside cancer markers in urine sample was realized.     

Light responses in the green sulfur bacterium Prosthecochloris aestuarii: changes in prosthecae length, ultrastructure, and antenna pigment composition

The morphology (mainly prosthecae length), ultrastructure, and antenna pigment composition of the green sulfur bacterium Prosthecochloris aestuarii changed when grown under different light intensities. At light intensities of 0.5 and 5 micromol quanta m(-2) s(-1), the cells had a star-like morphology. Prosthecae, the characteristic appendages of the genus Prosthecochloris, were 232 nm and 194 nm long, respectively. In contrast, when grown at 100 micromol quanta m(-2) s(-1), these appendages were shorter (98 nm) and the cells appeared more rod-shaped. Transmission electron microscopy revealed a significant decrease in the cell perimeter to area ratio and in the number of chlorosomes per linear microm of membrane as light intensity increased. In addition to these morphological and ultrastructural responses, Prosthecochloris aestuarii exhibited changes in its pigment composition as a function of light regime. Lower specific pigment content and synthesis rates were found in cultures grown at light intensities above 5 micromol quanta m(-2) s(-1). A blue shift in the bacteriochlorophyll (BChl) c Q(y) absorption maximum of up to 17.5 nm was observed under saturating light conditions (100 micromol quanta m(-2) s(-1)). This displacement was accompanied by changes in the composition of BChl c homologs and by a very low carotenoid content. The morphological, ultrastructural and functional changes exhibited by Prosthecochloris aestuarii revealed the strong light-response capacity of this bacterium to both high and low photon-flux densities.     

Surface imprinted beads for the recognition of human serum albumin

The synthesis of poly-aminophenylboronic acid (ABPA) imprinted beads for the recognition of the protein human serum albumin (HSA) is reported. In order to create homogeneous recognition sites, covalent immobilisation of the template HSA was exploited. The resulting imprinted beads were selective for HSA. The indirect imprinting factor (IF) calculated from supernatant was 1.6 and the direct IF, evaluated from the protein recovered from the beads, was 1.9. The binding capacity was 1.4 mg/g, which is comparable to commercially available affinity materials. The specificity of the HSA recognition was evaluated with competitive experiments, indicating a molar ratio 4.5/1 of competitor was necessary to displace half of the bound HSA. The recognition and binding of the imprinted beads was also tested with a complex sample, human serum and targeted removal of HSA without a loss of the other protein components was demonstrated. The easy preparation protocol of derivatised beads and a good protein recognition properties make the approach an attractive solution to analytical and bio-analytical problems in the field of biotechnology.     

An experimental and theoretical study of the morphine binding capacity and kinetics of an engineered opioid receptor

Electrochemical real-time monitoring of ligand binding to an engineered opioid receptor specific for morphine is reported. In the particular systems studied, 90% of the binding was found to be completed after only 85-120 s. Thus, the binding kinetics has proven to be more rapid than previously believed. The observed association rate constant for the morphine binding reaction was calculated to be 215 M(-1)s(-1). A theoretical analysis of the experimental binding data suggested that the binding sites of the engineered opioid receptor could best be described by a model having two populations of binding sites: K(D)=40 microM (13 micromol/g) and K(D)=205 microM (29 micromol/g). Furthermore, a theoretical model was developed in order to explain the observed binding of the engineered opioid receptor. This model suggested that the binding sites on the polymer surface are up to 5.1A deep and they allow 100% of the ligand (morphine) to anchor itself into the site. The predicted theoretical maximum binding capacity for the reported receptor is calculated to be approximately 2 mmol/g polymer (based on an increase of cavity density).     

Antioxidant capacity and polyphenolic components of teas: implications for altering in vivo antioxidant status

The Oxygen Radical Absorbance Capacity (ORAC) assay was used to determine the total antioxidant capacity of tea. Green and black teas (n = 18) had a mean antioxidant capacity of 761.1 +/- 85.3 micromol Trolox Equivalents (TE) per g dry matter. However, their antioxidant capacity varied from 235 micromol to over 1526 micromol Trolox equivalents (TE)/g dry matter, and total phenolics ranged from 32 to 147 mg/g in different commercial teas. One tea phenolics extract had an antioxidant capacity of 4796 micromol TE/g dry matter and 625 mg total phenolics/g. On a dry matter basis, an antioxidant capacity of 761 micromol TE/g is considerably higher than any of the other fruits and vegetables measured in our laboratory. However, since dry tea is not consumed directly, brewing conditions may influence the final antioxidant capacity in the tea as consumed. We tested both green and black teas by placing one tea bag (1.95 g) in 150 ml (5 oz.) of boiling water. In the first brewed cup, approximately 84% of the total antioxidant activity was solubilized within the first 5 min of brewing. An additional 13% of the antioxidant activity was extracted into the second glass of 150 ml with an additional 5 min of brewing. At the dilutions obtained after the first brewing, the tea as consumed would contain approximately 8. 31 micromol TE per ml. This total antioxidant capacity compares to other drinks from fruits and vegetables that had antioxidant capacity values ranging from 1.6 to 15 micromol TE/ml. At these antioxidant levels, consumption of 150 ml of tea could make a significant contribution to the total daily antioxidant capacity intake. (-)-Epicatechin and (+)-catechin, two components from tea, had an antioxidant capacity of 2.36 and 2.49 micromol/micromol or 8. 13 and 8.58 micromol/mg, respectively. In 16 tea samples we observed a mean of 10.0 +/- 0.6 micromol TE/mg total phenolics. Tea can be an important source of what has been referred to as "non-nutrient" antioxidant phytochemicals. However, with the variation that exists in antioxidant capacity with various tea preparations, measures of antioxidant capacity intake are critical to the study of intake and health outcomes and/or biomarkers of health outcomes.     

Molecularly imprinted thin film self-assembled on piezoelectric quartz crystal surface by the sol-gel process for protein recognition

A novel method of combining sol-gel and self-assembly technology to prepare a human serum albumin (HSA)-imprinted film on the surface of piezoelectric quartz crystal (PQC) Au-electrode modified with thioglycolic acid was described in this paper. The imprinting process was characterized by using the piezoelectric quartz crystal impedance (PQCI) technique and electrochemical impedance technique. Scanning electron microscope (SEM) was employed to characterize the surface morphology of the resultant imprinted film. The piezoelectric technique and electrochemical impedance technique were also employed to investigate the binding performance of the sol-gel-imprinted film with the template protein. The results showed that the imprinted PQC film can give selective recognition to the template protein. The effects of salts and solvents on the binding capacity of the imprinted film with protein were discussed in detail. Other influencing factors (temperature and pH) have also been investigated. This self-assembly sol-gel imprinting technique was proved to be an alternative method for the preparation of biomacromolecule-imprinted thin film.     

Chitosan-silica complex membranes from sulfonic acid functionalized silica nanoparticles for pervaporation dehydration of ethanol-water solutions

Nanosized silica particles with sulfonic acid groups (ST-GPE-S) were utilized as a cross-linker for chitosan to form a chitosan-silica complex membranes, which were applied to pervaporation dehydration of ethanol-water solutions. ST-GPE-S was obtained from reacting nanoscale silica particles with glycidyl phenyl ether, and subsequent sulfonation onto the attached phenyl groups. The chemical structure of the functionalized silica was characterized with FTIR, (1)H NMR, and energy-dispersive X-ray. Homogeneous dispersion of the silica particles in chitosan was observed with electronic microscopies, and the membranes obtained were considered as nanocomposites. The silica nanoparticles in the membranes served as spacers for polymer chains to provide extra space for water permeation, so as to bring high permeation rates to the complex membranes. With addition of 5 parts per hundred of functionalized silica into chitosan, the resulting membrane exhibited a separation factor of 919 and permeation flux of 410 g/(m(2) h) in pervaporation dehydration of 90 wt % ethanol aqueous solution at 70 degrees C.     

Studies on the possibility of histidine biosynthesis from histidinol, imidazolepyruvic acid, imidazoleacetic acid, and imidazolelactic acid by mixed ruminal bacteria, protozoa, and their mixture in vitro

The possibility of histidine (His) synthesis using a main biosynthetic pathway involving histidinol (HDL) and also the recycling capability of imidazolic compounds such as imidazolepyruvic acid (ImPA), imidazoleacetic acid (ImAA), and imidazolelactic acid (ImLA) to produce His were investigated using mixed ruminal bacteria (B), protozoa (P), and a mixture of both (BP) in an in vitro system. Rumen microorganisms were anaerobically incubated at 39 degrees C for 18 h with or without each substrate (2 mM) mentioned. His and other related compounds produced in both the supernatants and hydrolyzates of the incubation were analyzed by high-performance liquid chromatography. B, P, and BP suspensions failed to show His synthesizing ability when incubated with HDL. His was synthesized from ImPA by B, P, and BP. Expressed in units "per gram of microbial nitrogen (MN)", ImPA disappearance was greatest in B (72.7 micromol/g MN per hour), followed by BP (33.13 micromol/g MN per hour) and then P (18.6 micromol/g MN per hour) for the 18-h incubation period. The production of His from ImPA in B (240.0, 275.9, and 261.2 micromol/g MN in 6, 12, and 18 h incubation, respectively) was about 3.5 times higher than that in P (67.3, 83.8, and 72.7 micromol/g MN in 6, 12, and 18 h incubation, respectively). Other metabolites produced from ImPA were ImLA, ImAA, histamine (HTM), and urocanic acid (URA), found in all microbial suspensions. ImLA as a substrate remained without diminution in all microbial suspensions. Although ImAA was found to be degraded to a small extent (3.4-6.3%) only after 18 h incubation, neither His nor other metabolites were detected on the chromatograms. These results have been demonstrated for the first time in rumen microorganisms and suggest that His may be an essential amino acid for rumen microorganisms.     

Molecularly imprinted chitosan-genipin hydrogels with recognition capacity toward o-xylene

A molecularly imprinted material was developed from hydrogels of chitosan (CS) cross-linked with genipin (GNP) using o-xylene as the template molecule. Gelling time, mechanical, and diffusion properties of CS-GNP hydrogels were initially investigated to establish optimal conditions to prepare molecularly imprinted hydrogels (MIHs). The elastic modulus was found to be directly proportional to the degree of cross-linking (R = moles of genipin/moles of glucosamine) while the diffusion of water, as monitored by magnetic resonance imaging, decreased with R. CS-GNP hydrogels of varying R were imprinted with o-xylene (MIH(o-xylene)). The adsorption capacity of o-xylene by MIH(o-xylene) was greater than the corresponding control hydrogels, particularly at R = 0.25. Freundlich isotherms yielded a better fitness than Langmuir ones and afforded n and Q(max)values of 2.55 and 103.3 mg/g, respectively. The imprinted hydrogel showed the highest adsorption capacity for o-xylene; however, the material was not highly selective as it also exhibited the capacity to adsorb m- and p-xylene isomers. In turn, the MIH(o-xylene) showed a low adsorption when 2-fluorotoluene was used in rebinding experiments, suggesting that molecular recognition by the binding sites is influenced by the electronic and steric properties of the analyte molecule, thus effectively confirming the imprinting effect within the MIH(o-xylene) network. This work opens the possibility to future development of materials with the capacity to adsorb o-xylene analogue molecules such as contaminants bearing chlorinated aromatic structures.     

Binding site characteristics of 17beta-estradiol imprinted polymers

The variety of applications utilizing molecularly imprinted polymers (MIPs) requires synthetic strategies yielding different MIP formats including films, irregular particles, or spheres, along with precise knowledge on the specific material characteristics, such as binding capacity and binding efficiency of these materials. In response to this demand, MIPs are prepared in different formats by variation of the polymerization methodology. It is commonly agreed that micro- and sub-microspheres are particularly advantageous MIP formats, due to their monodispersity and facile synthesis procedures in contrast to conventional imprinted polymers prepared by bulk polymerization. However, the differences in actual rebinding characteristics of different MIP formats based on molecular interactions under a variety of binding/rebinding conditions have not been studied in detail to date. Consequently, the present work details an analytical strategy generically applicable to MIP systems for rebinding studies including equilibrium binding, non-equilibrium binding, and release experiments enabling more profound understanding on the molecular interactions between the imprinted materials and the template molecules. In this study, three MIP formats were considered for the same template molecule, 17beta-estradiol: irregularly shaped particulate polymers prepared by bulk polymerization and grinding, microspheres, and sub-microspheres. The latter two formats were synthesized via precipitation polymerization using different processing strategies. The morphologies and porosities of the resulting imprinted materials were characterized by scanning electron microscopy (SEM) and Brunauer-Emmett-Teller (BET) analysis, respectively. The obtained results indicate that microspheres prepared by precipitation polymerization provide superior rebinding properties during equilibrium binding in contrast to bulk polymers and sub-microspheres, and that the rebinding properties are different during equilibrium binding versus non-equilibrium binding. The median binding affinity constant determined during non-equilibrium rebinding is higher than the values obtained from equilibrium rebinding. Furthermore, the binding site distribution appears more homogeneous thief derived from non-equilibrium rebinding, as reflected in a heterogeneity index of m=0.725. Moreover, it is hypothesized that the specific interactions between template and monomers are related to the porosity of the imprinted polymers, which implies that the amount of binding sites and the pore sized distribution of the imprinted materials are a critical factor in achieving the desired MIP performance in various analytical applications. The BET results indicate that particles prepared with lower cross-linker-to-template ratio have a reduced surface area. Furthermore, it can be expected that there are less specific binding sites available at particles with reduced surface area and pore volume given similar distribution of the binding sites, as confirmed by the equilibrium binding isotherm studies. The pore size distribution results reveal that control of the pore size in the range of 100-180 A is essential to obtain the desired retention properties and Gaussian peak shape during HPLC analysis of small molecules.     

Computational predictions and experimental affinity distributions for a homovanillic acid molecularly imprinted polymer

Density Functional Theory calculations have been used to select, among a set of chemicals traditionally used in the formulation of non-covalent molecularly imprinted polymers (MIPs), the best functional monomer and porogenic solvent for the construction of a recognition element for the dopamine metabolite homovanillic acid (HVA). Theoretical predictions were confirmed through batch binding assays and voltammetric detection. The computational method predicts that trifluoromethacrylic acid and toluene are the monomer and solvent rendering the highest stabilization energy for the pre-polymerization adducts. HVA-MIP prepared using this formulation gives rise to a binding isotherm that is accurately modelled by the Freundlich isotherm. The binding properties of this polymer were estimated using affinity distribution analysis. An apparent number of sites of 13 micromol g(-1) with an average affinity constant of 2 x 10(4) M(-1) was obtained in the concentration window studied.     

Design and synthesis of molecularly imprinted polypyrrole based on nanoreactor SBA-15 for recognition of ascorbic acid

Molecular imprinting is an attractive technique for preparing mimics of natural and biological receptors. Nevertheless, molecular imprinting for aqueous systems remains a challenge due to the hydrogen bonding between templates and functional monomers destroyed in the bulk water. The hydrogen bonding between templates and monomers are the most crucial factor governing recognition, particularly in non-covalent molecularly imprinted polymers. Using mesoporous materials for molecular imprinting is an effective approach to overcome this barrier and to remove the limitations of the traditional molecularly imprinted polymers which include incomplete template removal, small binding capacity, slow mass transfer, and irregular materials shape. Here, SBA-15 was used as a mesoporous silica material for synthesis of molecularly imprinted polypyrrole. The pyrrole monomers and template molecules were immobilized onto the SBA-15 hexagonal channels, and then polymerization occurred. The resulting nanocomposites were characterized by Fourier transform infrared (FT-IR) analysis, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) methods. In batch rebinding tests, the imprinted nanocomposites reached saturated adsorption within 100min and exhibited significant specific recognition toward the ascorbic acid (AA) with high adsorption capacity (83.7mgg(-1)). To further illustrate the recognition property of the imprinted nanocomposites, binary competitive and non-competitive adsorption experiments were performed with ascorbic acid, dopamine, paracetamol and epinephrine. The imprinting factors for these compounds in non-competitive adsorption experiments were 3.2, 1.5, 1.4 and 1.3, respectively. The results showed that the imprinted nanocomposites exhibited significant adsorption selectivity for the ascorbic acid against the related compounds.     

Adsorption of papain with Cibacron Blue F3GA carrying chitosan-coated nylon affinity membranes

Covalent coupling of chitosan (CS) to activated nylon membrane was performed after the reaction of the microporous nylon membrane with formaldehyde. Non-specific adsorption on the CS-coated nylon membrane decreased greatly, compared with plain nylon membrane. The dye Cibacron Blue F3GA (CB F3GA) as a ligand was then covalently immobilized on the CS-coated membranes. Physical properties of the composite membrane and its applications in affinity membrane chromatography were examined. The contents of CS and CB F3GA-attached membranes were 89.6 mg/g nylon membrane and 146.1 micromol/g nylon membrane, respectively. These CB F3GA-attached composite membranes were used in the papain adsorption studies. Higher papain adsorption capacity, up to 235.3mg/g affinity membrane, was obtained. The adsorption isotherm fitted the Freundlich model well. Significant amount of the adsorbed papain (about 94.3%) was eluted by 1.0M NaSCN at pH 9.0. Experiments on regeneration and dynamic adsorption were also performed. It appears that CB F3GA-CS nylon membranes can be applied for papain separation without causing any denaturation.