Prediction of chromatographic retention, pKa values and optimization of the separation of polyphenolic acids in strawberries.

Polyphenolic acids are a complex group of compounds that have attracted enormous attention in the last few years because of their biological properties. In this work, the proportion of organic modifier and the pH of acetonitrile-water mixtures used as mobile phases were optimized in order to separate a series of polyphenolic compounds. The linear solvation energy relationship formalism based on the single solvent polarity parameter, E(T)N was used to predict their chromatographic behavior as a function of the percentage of acetonitrile in the eluent. Moreover, the correlation established between retention and the pH of the aqueous-organic mobile phase was used to optimize the pH of the mobile phase. The optimized mobile phase is composed of acetonitrile and formic acid buffer adjusted to pH 4.25, with 12% (v/v) acetonitrile. Also, the pKa values of polyphenolic acids in acetonitrile-water mixtures were determined using chromatographic data, and in order to validate the optimized conditions, a series of polyphenolic compounds was studied in strawberries.

A soft-modeling approach to interpret thermodynamic and conformational transitions of polynucleotides.

Multivariate outputs from the experimental monitoring of biochemical processes are usually difficult to interpret applying methods based on a priori chemical models. Curve resolution methods are model-free procedures, generally known as soft-modeling methods, which obtain the concentration profiles and instrumental responses of each individual species involved in a multivariate monitored process without making any kind of external assumption. Of the curve resolution methods available, the alternating least squares (ALS) is proposed here because of its ability to operate on one or on several matrices. Furthermore, ALS allows the introduction of information related to the internal data structure and to the general features of the concentration profiles and instrumental responses through the input of suitable constraints in the iterative resolution procedure. The ALS potential is tested on several data sets coming from the multivariate spectrometric monitoring of polyuridylic (polyU), polycytidylic (polyC), and polyadenylic (polyA) protonation equilibria in dioxane/water 30% (v/v). Information concerning the evolution of the concentration profiles and the spectra of each individual species involved in the acid-base equilibria, the presence and pattern of polyelectrolyte effects, and the presence of conformational transitions associated or not with the proton uptake process is presented.

Three-way curve resolution applied to the study of solvent effect on the thermodynamic and conformational transitions related to the protonation of polycytidylic acid.

Solvent effect on the acid-base behavior of polycytidylic acid [poly(C)] is studied by means of potentiometric and spectrometric (UV and CD) procedures. Low-polarity biological environments are mimicked by using a 30% (v/v) dioxane-water mixture. Experiments performed in this medium are compared with previously reported results in aqueous solution in order to determine changes in both thermodynamic and structural aspects associated with modifications of the biomolecular surroundings. Potentiometric and spectrometric studies reveal the presence of a nonlinear polyelectrolytic effect associated with the protonation of poly(C) in the hydroorganic mixture, different from the analogous effect in aqueous solution. The curve resolution method alternating least squares is applied to the poly(C) spectrometric data. Concentration profiles and spectra of both deprotonated poly(C) and half-protonated [poly(C).poly(CH+)] are thus obtained. The fully protonated species poly(CH+) precipitates in the hydroorganic medium at pH values lower than 4. Evidence for the ordered structure of both poly(C) and [poly(C).poly(CH+)] species is seen through the comparison of the macromolecule spectra with those of the cytidine-3',5'-cyclic monophosphate monomer. Polarity decreases around the macromolecule produce significant hypochromicity in the CD spectra and hyperchromicity in the UV spectra, both signs of a disordering effect in the macromolecular structure due to the weakening of base stacking interactions.

Application of a self-modeling curve resolution approach to the study of solvent effects on the acid-base and copper (II)-complexing behavior of polyuridylic acid.

The solvent effect on the acid-base and complexation behavior of the homopolynucleotide polyuridylic acid (poly(U)) has been studied by means of potentiometric and spectrometric titrations (circular dichroism and UV-VIS) in water and in 30 and 50% (v/v) dioxane-water media. The potentiometric studies revealed the absence of polyelectrolytic effects in the acid-base equilibrium, and the spectrometric experiments detected only a random coil conformation associated with both the protonated and deprotonated species. The common behavior observed in the three media seems to indicate the weakness of both intramolecular interactions, i.e., base stacking, and solute/solvent interactions, i.e., hydrogen-bonding, and consequently their small effect during the protonation process. Differences regarding the solubility of the deprotonated species in the solvents used are due to the difficult stabilization of such a charged species in the low polar environment of the dioxane-water mixtures. Complexation has been exhaustively studied in aqueous media, and no conformational changes have been noticed in the only copper(II)-poly(U) complex detected. The inclusion of the copper(II) ion in the macromolecular skeleton of the polynucleotide does not contribute to an ordination of the structure, which remains as a random coil. No comparison between this equilibrium in aqueous solution and in the hydroorganic mixtures could be carried out since the limited pH range of the soluble complex in those solvent mixtures prevented a rigorous quantitative monitoring of such a chemical process.