Studies on interpolymer self-organisation behaviour of protoporphyrin IX bound poly(carboxylic acid)s with complimentary polymers by means of fluorescence techniques

Covalently bound protoporphyrin IX was used as a fluorophore to investigate the interpolymer complex formation between the poly(carboxylic acid)s, PMAA/PAA and poly(N-vinyl pyrrolidone), PVP, poly(ethylene oxide), PEO or poly(ethylene glycol), PEG. Absorption and emission spectral properties of protoporphyrin IX bound to PAA, PMAA and PVP have been studied. Protoporphyrin IX in poly(MAA-co-PPIX) was found to be present in the dimer or higher aggregated form at low pH due to the environmental restriction imposed by the polymer whereas in the case of poly(AA-co-PPIX) and poly(VP-co-PPIX), PPIX exists in monomeric form. The fluorescence intensity and lifetime of PPIX bound to poly(carboxylic acid)s increase on complexation through hydrogen bonding with PVP, PEO and PEG due to the displacement of water molecules in the vicinity of the PPIX. Poly(MAA-co-PPIX) shows longer fluorescence lifetime due to the more compact interpolymer complexation as compared to poly(AA-co-PPIX) due to the enhanced hydrophobicity of PMAA. Poly(VP-co-PPIX) shows a decrease in the fluorescence lifetime on complexation with PMAA or PAA due to the hydrophilic and microgel like environment of the fluorophore bound to PVP. The contrasting behaviour of the same polymer adduct with respect to the site of the fluorophore is interpreted to be due to the solvent structure which determines the environment of the fluorophore.     

Interaction between poly(N-vinylpyrrolidone) and poly(acrylic acid)s: Influence of hydrogen and cupric ions on the adduct formation

Interpolymer adduct formation between poly(N-vinylpyrrolidone) (PVP) and poly(methacrylic acid) (PMAA) is mainly due to hydrogen bonding. It is found that the interpolymer adduct formation is enhanced in the presence of Cu(II). A simple turbidity measurement making use of a spectrophotofluorometer is described for the study of the interpolymer adduct formation. Enhanced adduct formation in the presence of Cu(II) is described by the empirical relation d[PAd]/D[PVP] = k × 10^4α[Cu(II)]^α, where PAd represents the interpolymer adduct and α and k are constants. Similar results have been obtained in the case of interpolymer adduct formation between poly(acrylic acid) (PAA) and PVP. In the above expression α signifies the influence of chelation on Cu(II)–PAA/PMAA–PVP-type complex formation and k is the extent of PVP–PAA/PMAA interaction. The enhancement of adduct formation in the presence of Cu(II) is more in PAA than in PMAA. Turbidity and viscosity measurements further indicate that the influence of Cu(II) on interpolymer adduct formation between PVP and PMAA or PAA is more in the case of PAA than PMAA, as PAA is a better chelating ligand. But the extent of adduct formation is more in the case of PMAA in the absence of Cu(II) ions due to hydrophobic interactions exerted by methyl groups.     

LASER FLASH SPECTROSCOPY OF METHYLENE BLUE WITH NUCLEIC ACIDS. EFFECTS OF IONIC STRENGTH AND pH

Laser flash spectroscopic measurements were made on methylene blue complexed to the synthetic polynucleotides poly[d(G-C)] and poly[d(A-T)] in solutions of varying ionic strength and pH. Triplet decay rates and rates of triplet quenching by oxygen have been measured for the polymer/dye solutions. The triplet decay and oxygen quenching rates of methylene blue in complex with poly[d(A-T)] are much less sensitive than those with poly[d(G-C)] with respect to variations in ionic strength. It is also shown that protonation of the triplet state of MB+ with poly[d(A-T)] is slower than that of free dye. These results indicate strong binding of the dye to poly[d(A-T)]. Excitation of the MB+/poly[d(A-T)] complex at 665 nm yielded single exponential decay kinetics, in contrast with excitation at 600 nm where double exponential kinetics were measured. This is tentatively assigned to excitation of dye dimer bound to this polymer.     

Preparation and complexation of an A–B–A type triblock copolymer consisting of helical poly(L-proline) and random-coil poly(ethylene oxide)

By using L -proline N-carboxyanhydride (LPNCA) and amino-group terminated poly(ethylene oxide) (ATPEO), an A–B–A–type [A = poly(L -proline) (PLP), B = poly(ethylene oxide) (PEO)] triblock copolymer (POP) was prepared which is water-soluble. In the POP, A = PLP is helical, and B = PEO is random coil. From the observations of the NMR spectra, specific optical rotation, and x-ray diffraction of the POP, it was found that the PLP component of the POP exists nearly as Form II PLP with trans-configuration, and interferes the crystal growth of PEO component, in solid state. With the addition of PMAA into an aqueous POP solution, dramatic decreases of reduced viscosity and pH are observed until the unit-mole-concentration-ratio (UMCR) [PMAA]/[POP] reaches its value of unity, while a distinct increase in turbidity appears. This shows a 1 : 1 interpolymer complex formation between PMAA and POP in aqueous medium through hydrogen bonding. The curves of viscosity, pH, and turbidity versus UMCR [PMAA]/[POP] show breaks at [PMAA]/[POP] = 0.3, suggesting the selective complexation of PLP component (ca. 30 unit-mol %) of POP with PMAA. The x-ray diffraction curve of the complex POP/PMAA shows entirely no diffraction patterns, indicating that the ordered POP structure (mainly due to that of PLP component) is completely destroyed owing to the complexation between POP and PMAA.     

PH effects in the complex formation and blending of poly(acrylic acid) with poly(ethylene oxide)

The effect of pH on the complex formation between poly(acrylic acid) (PAA) and poly(ethylene oxide) (PEO) has been studied in aqueous solutions by turbidimetric and fluorescent methods. It was shown that the formation of insoluble interpolymer complexes is observed below a certain critical pH of complexation (pH(crit1)). The formation of hydrophilic interpolymer associates is possible above pH(crit1) and below a certain pH(crit2). The effects of polymer concentrations in solution and PEO molecular weight as well as inorganic salt addition on these critical pH values were studied. The polymeric films based on blends of PAA and PEO were prepared by casting from aqueous solutions with different pHs. These films were characterized by light transmittance measurements and differential scanning calorimetry. The existence of the pH value above which the polymers form an immiscible blend was demonstrated. The transitions between the interpolymer complex, miscible blend, and immiscible blend caused by pH changes are discussed. The recommendations for preparation of homogeneous miscible films based on compositions of poly(carboxylic acids) and various nonionic water-soluble polymers are presented.     

Azole-linked coumarin dyes as fluorescence probes of domain-forming polymers.

The binding of 7-aminocoumarins, substituted in the 3-position with heterocyclic benzimidazole or benzothiazole groups by domain-forming polymers in water has been studied. The acrylic polyelectrolyte, poly(methacrylic) acid (PMAA) was used as a solubilizing agent for coumarin dyes 6, 7, and 30 in water. The acid-base properties of these bound coumarin dyes were monitored spectroscopically on titration of aqueous solutions. Alterations in the fluorescence wavelength and intensity, quantum yields, lifetimes, and polarization are consistent with the preferential binding of the dyes in compact hydrophobic domains that form at a pH regime in which the polymer is in its protonated (uncharged) state. In this pH range (<4.0), coumarins 7 and 30 are bound as monocations, whereas coumarin 6 remains in its neutral form. Reduced quantum yields and lifetimes of fluorescence for cationic coumarins can be understood in terms of the imposition of a low-lying electron transfer state, an example of a twisted intramolecular charge transfer (TICT) intermediate. Effects of polymer microenvironment on the rate of TICT state decay (a reverse electron transfer) are observed. Coumarins of the azole type may find use as fluoroprobes of the microenvironments of proteins and other biological macromolecules and as agents for pH sensing.     

Interpolymer complex between poly(ethylene oxide) and poly(carboxylic acid)

An interpolymer complex was prepared by mixing aqueous solutions of poly(ethylene oxide) (PEO) and of a poly(carboxylic acid), i.e., poly(acrylic acid)(PAA), poly(methacrylic acid)(PMAA), or styrene-maleic acid copolymer(PSMA). The complexation mechanism was discussed on the basis of results of such experimental methods as viscosity, potentiometric titration, and turbidimetry. The hydrogen bond is primarily involved in these complexations, but the influence of hydrophobic interaction on complexation can not be ignored. If the degree of dissociation α of carboxylic acid or the degree of polymerization P_n of PEO was perceptibly changed, a stable complex was obtained at about α 0.1 or P_n (PEO) = 40 for PMAA, 200 for PAA. This fact indicates that more than a definite number of binding sites are necessary for a stable interpolymer complex to be formed and that cooperative interaction among active sites plays an important role in complex formation.     

Studies on the dynamics of poly(carboxylic acids) with covalently bound thionine and phenosafranine in dilute aqueous solutions

The poly(carboxylic acid) bound phenosafranine and thionine dyes show that, the fluorescence intensity and lifetime increases first and starts to decrease after reaching a maximum at pH 4.0. The fluorescence decay curve of the fluorophore bound polymers follow the biexponential decay fit independent of pH, while poly(MAA-Th) follows single exponential function above pH 4.0. At low pH, a more compact environment of the fluorophore exerts a more hydrophobic environment. In the subnanosecond time domain the solvation process is found to be incomplete while in the nanosecond time scale the solvation of the macromolecular chains is found to be over. The time resolved fluorescence spectra of the polymer bound fluorophores at different pH indicate distinct hydrophobic and hydrophilic environments due to the dynamics of the macromolecules in dilute aqueous solutions. For the first time structural transitions involving solvent are observed in the nanosecond and picosecond time domains for the same macromolecule.     

Immobilization of polymer-protected metal colloid catalysts by the formation of polymer hydrogen bond complexes

poly(N-vinyl-2-pyrrolidone)-(PVP) and polyvinyl alcohol-(PVA) protected nanoscopic noble metal colloidal catalysts were immobilized to prepare the corresponding heterogeneous catalysts by forming polymer hydrogen bond complexes of polyacrylic acid (PAA) with PVP or PVA. A PVP–PAA–Pd catalyst was found to be very active and selective for the partial hydrogenation of cyclopentadiene to cyclopentene.     

Fluorescence correlation spectroscopy studies of diffusion of a weak polyelectrolyte in aqueous solutions

We apply fluorescent correlation spectroscopy (FCS) to investigate solution dynamics of a synthetic polyelectrolyte, i.e., a weak polycarboxylic acid in aqueous solutions. The technique brings single molecule sensitivity and molecular specificity to dynamic measurements of polyelectrolyte solutions. Translational diffusion of Alexa-labeled poly(methacrylic acid), PMAA*, chains was studied in very dilute, 10(-4) mg/ml, solutions as a function of solution pH and ionic strength. The observed changes in diffusion coefficients were consistent with about twofold expansion of PMAA* coils when pH was changed from 5 to 8, and with chain contraction for alkaline metal ion concentrations from 0.01 to 0.1 M. The dependence of the hydrodynamic size of PMAA* chains on the counterion type followed the sequence: Li(+)>Na(+) approximately equal to Cs(+)>K(+). The dependence of translational diffusion on polyacid concentration was weak at the low concentration limit, but chain motions were significantly slower at higher polymer concentrations when PMAA chains overlapped. Finally, measurements of dynamics of PMAA* chains in "salt-free" solutions showed that self-diffusion of PMAA* chains significantly slowed down when PMAA concentration was increased, probably reflecting the sensitivity of PMAA* translational motions to the onset of interchain domain formation. These results illustrate the utility of the FCS technique for studying hydrodynamic sizes of polyelectrolyte coils in response to variation in solution pH or concentration of salt and polyelectrolytes. They also suggest that FCS will be a promising technique for selective observation of the dynamics of polyelectrolyte components in complex polymer mixtures.     

Interpolymer complex formation between helical poly(L-proline) and random-coil poly(methacrylic acid) through hydrogen bonding

Interpolymer complex formation between poly(L -proline) (PLP) with helical structure and poly(methacrylic acid) (PMAA) with random-coil structure through hydrogen bonding in aqueous medium has been studied by several experimental techniques, e.g., viscometry, turbidimetry, potentiometry, conductometry, scanning electron microscopy, and x-ray diffraction methods. The decreases in reduced viscosity of the solution on addition of an increasing quantity of PLP to a constant amount of PMAA reveals the formation of a complex between PLP and PMAA. The minimum in reduced viscosity at a unit-mole ratio [PLP]/[PMAA] = 1.0 suggests a 1 : 1 complex formation. A distinct change in the curves for turbidity, pH, and conductance versus [PLP]/[PMAA] supports this conclusion. A scanning electron micrograph for the 1 : 1 PLP–PMAA complexes shows that the PLP/PMAA complex has the shape of entangled long fibers. An x-ray diffraction pattern for the PLP/PMAA complexes gives no diffraction patterns which appear in pure PLP, indicating the destruction of the helical structure of PLP due to the interpolymer complexation. Mixtures of PMAA with poly(γ-hydroxy-L -proline) (PHLP) which has a similar conformation as PLP, but involves intra- or intermolecular hydrogen bonds, has also been investigated by vicometry measurements. The reduced viscosity of a solution of the mixed polymers increases with increasing [PHLP], indicating no complex formation. All the results reveal that the magnitude and the nature of the forces acting in the polymers play an important role in interpolymer complexation.     

Light Scattering Study of the Antibody–Poly(methacrylic acid) and Antibody–Poly(acrylic acid) Conjugates in Aqueous Solutions

The effect of the conformational state of the polymer coil on the properties of protein–polymer conjugates has been studied for the conjugates of antibody (monoclonal antibody from 6C5 clone against inactivated rabbit muscle glyceraldehyde‐3‐phosphate dehydrogenase; Ab) with poly(methacrylic acid) (PMAA) or poly‐(acrylic acid) (PAA). The pH‐dependencies of molecular properties and structural parameters of aqueous solutions (radius of gyration, intensity of scattered light, hydrodynamic diameter, and polydispersity index) of Ab, PMAA, and PAA and their conjugates, i. e., Ab‐PMAA and Ab‐PAA, have been studied using static and dynamic light scattering techniques. While free Ab aggregates in solution and precipitates at its isoelectric point, the covalent attachment of a charged polymer to Ab prevents its association and shifts the precipitation point towards more acidic values (from pH 5.95 for Ab to pH ˜ 4.8 for Ab‐PMAA). The predominant role of the conformational status of the polymer in the process of conjugate precipitation has been considered. Contrary to the precipitation of Ab‐PMAA, the formation of stable colloidal particles was suggested for Ab‐PAA at pH < 4.8. In the conjugates, polymer chains surround the protein globule in an extremely compact manner while Ab significantly affects the polymer conformation. The essentially larger hydrodynamic radii of conjugates, when compared with their radii of gyration, confirm the strong interaction of conjugates with solvent molecules.     

Surface Properties of Polyacid–Poly(N-Vinyl Pyrrolidone) Complexes

The effect of the complexation of polyacrylic (PAA) and polymethacrylic (PMAA) acids with poly(N-vinyl pyrrolidone) (PVP) on their surface properties was studied by measuring surface tension, conductivity, and the viscosity of aqueous mixed polymer solutions, as well as by the potentiometric titration at 293.15 K. The values of relaxation times and the surface activity of polycomplexes were calculated from kinetic data and the surface tension isotherms of the solutions. The complexation was found to increase the surface layer relaxation time, surface activity, and the ability of the macromolecules to reduce the surface tension of solvent. The values of adsorption free energy were calculated for PVP and polycomplexes. They were equal to –22.0, –23.0, and –24.8 kJ/base-mole (per one mole of PVP monomer unit) for PVP and PMAA–PVP and PAA–PVP polycomplexes, respectively.     

pH effects on the complexation, miscibility and radiation-induced crosslinking in poly(acrylic acid)-poly(vinyl alcohol) blends

The effect of pH on the complexation of poly(acrylic acid) with poly(vinyl alcohol) in aqueous solution, the miscibility of these polymers in the solid state and the possibility for crosslinking the blends using gamma radiation has been studied. It is demonstrated that the complexation ability of poly(vinyl alcohol) with respect to poly(acrylic acid) is relatively low in comparison with some other synthetic non-ionic polymers. The precipitation of interpolymer complexes was observed below the critical pH of complexation (pH(crit1)), which characterizes the transition between a compact hydrophobic polycomplex and an extended hydrophilic interpolymer associate. Films prepared by casting from aqueous solutions at different pH values exhibited a transition from miscibility to immiscibility at a certain critical pH, pH(crit2), above which hydrogen bonding is prevented. It is shown here that gamma radiation crosslinking of solid blends is efficient and only results in the formation of hydrogel films for blends prepared between pH(crit1) and pH(crit2). The yield of the gel fraction and the swelling properties of the films depended on the absorbed radiation dose and the polymer ratio. [Diagram: see text] SEM image of an equimolar PAA-PVA blend cast from a pH 4.6 solution.     

Characterization of poly(carboxylic acid)/poly(ethylene oxide) blends formed through hydrogen bonding by spectroscopic and calorimetric analyses

The solid state of the complex between poly(acrylic acid) (PAA) and poly(ethylene oxide) (PEO), and that between poly(methacrylic acid) (PMAA) and PEO formed via hydrogen-bonding was studied by differential-scanning calorimetric (DSC) and by Fourier-transform infrared (FT–IR) spectroscopic measurements. Melting temperature T_m and the degree of the crystallinity X_c of PEO in the systems PAA (or PMAA)/PEO blends obtained from aqueous or dimethyl sulfoxide (DMSO) medium were measured in various unit mol % of PEO ([PEO]100/{[PAA(or PMAA)] + [PEO]}) where [ ] is the unit mole concentration. It was found that 50 unit mol % of PEO is a critical composition, which gives new evidence for the 1 : 1 complex formation between PAA (or PMAA) and PEO. From the FT–IR spectroscopic analysis in conjunction with DSC measurements we also found that the effects of solvent and of hydrophobic interaction (due to the α-methyl group of PMAA) are the important factors controlling the complexation in the solution and solid systems. These factors also affect the crystallization behavior and the microstructure of the PAA (or PMAA)/PEO blend in solid state.     

Photophysical investigations of interpolymer interactions in solutions of a pyrene substituted poly(acrylic acid), poly(vinyl amine), poly(1-aminoacrylic acid), and poly(1-acetylaminoacrylic acid)

Photophysical properties of the pyrene chromophore covalently bound to poly(acrylic acid) were used to investigate the interactions of a pyrene substituted poly(acrylic acid) (1) with poly(vinyl amine hydrochloride) (PVAm), poly(1-aminoacrylic acid) (PDA), and poly(1-acetylaminoacrylic acid) (PADA) in aqueous solutions. A number of photophysical parameters were obtained from fluorescence emission and excitation spectra, the deconvolution of decay curves for pyrene monomer, and excited state complex fluorescence and the quenching of pyrene monomer fluorescence by nitromethane in polymer solutions. These photophysical parameters were considered to reflect the inter- and intrapolymer interactions in solutions of 1 , PVAm, PDA, and PADA. The formation of interpolymer complexes between 1 and PVAm was noticed at low (< 4) as well as high (> 8) values, whereas PDA and 1 formed interpolymer complexes at low pH only. No interpolymer complex formation was detected in solutions of 1 and PADA under low or high pH conditions. The structures of interpolymer complexes formed between 1 and PVAm under low and high pH conditions were found to be determined by the conformation of 1 . There were significant differences in the interpolymer interactions of 1 and PDA in comparison to those of 1 and PVAm; in particular, the fluorescence from the excited state complex was enhanced in solutions of 1 and PVAm but quenched in solutions of 1 and PDA. The investigations of terpolymer solutions of 1 , PVAm, and PADA indicated that the nature of interpolymer complexes formed in terpolymer solutions was determined by Coulombic interactions of the amino and carboxylic group containing polymers.     

Release of a dye from hydrogen-bonded and electrostatically assembled polymer films triggered by adsorption of a polyelectrolyte

The absorption of dyes within hydrogen-bonded and electrostatically assembled multilayers and subsequent release of the dyes from the films were studied in situ using FTIR-ATR. Multilayers were composed of poly(methacrylic acid), PMAA, and poly(ethylene oxide), PEO (hydrogen-bonded multilayers), or of PMAA and 22% quarternized copolymer of N-ethyl-4-vinylpyridium bromide and 4-vinylpyridine, Q22 (electrostatically stabilized multilayers). After multilayer deposition, the solution pH was changed to produce excess charge within the films. Dyes with charge opposite to the excess charge of the film (Rhodamine 6G for hydrogen-bonded films or Bromophenol Blue for electrostatically assembled multilayers) were then allowed to absorb within multilayers. In both systems, the dyes were uniformly included within the films. The top layers largely affected the loading capacity of the multilayers, suggesting weaker binding of the dyes with the top layers. Dye release into a 0.01 M phosphate buffer was significantly smaller as compared to release in the presence of 0.05-0.5 mg/mL solutions of adsorbing polymers whose charge was the same as the excess charge within the films. We found that with the PMAA/PEO films, dye release did not depend on the concentration of polymer in solution, but was largely controlled by the amount of charge accumulated within the adsorbing polymer layer on the top of the film. For electrostatically stabilized PMAA/Q22 systems, dye release increased with increasing concentration of Q22 in solution, suggesting a significant contribution of the competition of solution species in the release mechanism. Our findings contribute to the understanding of interactions of small molecules with polymer multilayers and might have ramifications for novel applications of multilayer films as new materials for the controlled delivery of chemicals.     

Fluorescence and viscometry study of complexation of poly(acrylic acid) with poly(acrylamide) and hydrolysed poly(acrylamide)

Interpolymer complexation of poly(acrylic acid) with poly(acrylamide) and hydrolysed poly(acrylamide) was studied by fluorescence spectroscopy and viscometry in dilute aqueous solutions. Changes in chain conformation and flexibility due to the interpolymer association are reflected in the intramolecular excimer fluorescence of pyrene groups covalently attached to the polymer chain. Both poly(acrylamide) and hydrolysed poly(acrylamide) form stable complexes with poly(acrylic acid) at low pH. The molecular weight of poly(acrylic acid) and solution properties such as pH and ionic strength were found to influence the stability and the structure of the complexes. In addition, the polymer solutions mixing time showed an effect on the mean stoichiometry of the complex. The intrinsic viscosity of the solutions of mixed polymers at low pH suggested a compact polymer structure for the complex.     

Tuning the pH sensitivity of poly(methacrylic acid) brushes

The pH-induced swelling and collapse of surface-tethered, weak polyelectrolyte brushes is of interest for the development of actuators or to allow pH controlled transport or adsorption. This contribution discusses results of an extensive series of quartz crystal microbalance (QCM) experiments that aimed at (i) further understanding the influence of brush thickness and density on the pH responsiveness of poly(methacrylic acid) (PMAA) brushes and (ii) developing strategies that allow one to engineer the pH responsiveness and dynamic response range of PMAA based brushes. It was observed that, due to their high grafting density, the apparent pK(a) of surface-tethered PMAA differs from that of the corresponding free polymer in solution and also covers a broader pH range. The pK(a) of the PMAA brushes was found to depend on both brush thickness and density; thicker brushes showed a higher pK(a) value, and brushes of higher density started to swell at higher pH. The second part of the paper demonstrates the feasibility of the N-hydroxysuccinimide-mediated post-polymerization modification to engineer the pH responsiveness of the PMAA brushes. By using appropriate amine functionalized acids, it was possible to tune both the pH of maximum response as well as the dynamic response range of these PMAA based polyelectrolyte brushes.     

Photostability of pseudoisocyanine J aggregates in polymer films and ways of its improvement

The photostability of J aggregates of the pseudoisocyanine dye in polyvinylpyrrolidone (PVP) and poly(vinyl alcohol) (PVA) polymer films under continuous irradiation with visible light was investigated. The decay of J aggregates in PVP is predominantly due to the reaction of photooxidation with singlet oxygen generated by triplet-state pseudoisocyanine dimers. Unlike the case of PVP, J aggregates in PVA demonstrate significantly higher photostability because of a lower PVP permeability for oxygen. A significant improvement in the photostability of J aggregates is achieved by the introduction of scavengers of singlet oxygen into the polymer films.