Objective interpretation of ultraviolet-induced luminescence for characterizing pictorial materials.

Ultraviolet-induced Luminescence (UVL) is the property of some materials of emitting light once illuminated by a source of UV radiation. This feature is characteristic of some mediums and pigments, such as some red lakes, widely used for the realisation of works of art. On the one hand, UVL represents a like strike for a researcher in the cultural heritage field: in fact, UVL allows to characterise the state of conservation of the paintings and, in some cases, to recognize at glance some of the materials used by the artists. On the other hand, the contribution of UVL to the study of the artefacts is almost always limited to qualitative observation, while any speculation about the cause of the luminescence emission relies on the observer's expertise. The aim of this paper is to overcome this paradigm, moving a step toward a more quantitative interpretation of the luminescence signal. The obtained results concern the case study of pictorial materials by Giuseppe Pellizza da Volpedo (1868-1907, Volpedo, AL, Italy) including his iconic masterpiece Quarto Stato (1889-1901), but the method has general validity and can be applied whenever the appropriate experimental conditions occur. Once designed an appropriate set-up, the statistical comparison between the acquisitions performed on Quarto Stato, on a palette belonged to the master, on drafts made by the author himself and on a set of ad hoc prepared samples both with commercial contemporary pigments and prepared with the traditional recipe, shed some light on which materials have been employed by the artist, where they have been applied and support some intriguing speculations on the use of the industrial lakes in the Quarto Stato painting.

Gel polymerization in detergents: conversion efficiency of methylene blue vs. persulfate catalysis, as investigated by capillary zone electrophoresis.

Four types of detergents are commonly used in biochemical analysis of proteins and polypeptides: neutral, e.g., Triton X-100, Nonidet P-40; anionic, typically sodium dodecyl sulfate (SDS); cationic, e.g., cetyltrimethylammonium bromide CTAB), and zwitterionic, e.g., sulfobetaine 3-12 and 3[(3-cholamido-propyl) dimethyl-ammonio]-1-propane sulfonate (CHAPS). These detergents are utilized not only in the protein solubilization step, but also in the polyacrylamide gel matrix in which subsequent electrophoretic separation is carried out. The conversion efficiency of monomers into the growing polymer, in the presence of the four types of detergents, was assessed by capillary zone electrophoresis (CZE) in a micellar system comprising 100 mM SDS, by extracting unreacted monomers from the gel phase and quantifying the peaks separated by CZE. Two different catalyst systems were evaluated: the standard persulfate-N,N,N',N'-tetramethylethylenediamine (TEMED) couple and a mixture comprising methylene blue in presence of the redox couple sodium toluene sulfinate and diphenyliodonium chloride. In the chemically initiated system (persulfate), there was a strong inhibition of polymerization, decreasing in the following order: CTAB > sulfobetaine 3-12 > SDS > CHAPS > Triton X-100 (e.g., in 10 mM CTAB 100% inhibition was experienced). On the contrary, in photopolymerization (as driven by methylene blue) good conversion of monomers into the growing polymer (> 95%) was always obtained, independent of the nature of the detergent present in the polymerization step.(ABSTRACT TRUNCATED AT 250 WORDS)

On the efficiency of methylene blue versus persulfate catalysis of polyacrylamide gels, as investigated by capillary zone electrophoresis.

The efficiency of a novel method of photopolymerization, based on photoinitiating the reaction with methylene blue (MB), in presence of a redox couple (sodium toluenesulfinate and diphenyliodonium chloride), vs, the conventional persulfate--N,N,N',N'-tetramethylethylenediamine redox couple was investigated as a function of different effectors in solution. Oxygen dissolved in the gelling mixture strongly quenches persulfate catalysis, while leaving essentially unaltered the process initiated by photopolymerization. On the contrary, the presence of 8 M urea substantially accelerates a persulfate-driven reaction, boosting the conversion of monomers to near completion (> 98%) while leaving the photopolymerization process largely unaffected. Polyacrylamide polymerization has also been performed in a number of hydroorganic solvents (all in a 50:50 v/v ratio): dimethyl sulfoxide (DMSO), tetramethylurea, formamide and dimethylformamide. In all cases, the persulfate-catalyzed reaction was strongly quenched and even completely inhibited (in DMSO), whereas the photopolymerization process was essentially unaffected by any of these organic solvents. The reaction kinetics of the methylene blue-driven reaction could not be ameliorated when admixing an anionic dye (e.g., eosin Y) to the cationic MB, even when amply changing their molar ratios. Thus, it appears that photocatalysis with MB (and the redox couple sodium toluene-sulfinate and diphenyliodonium chloride) is a unique process, proceeding at optimum rate under the most adverse conditions, completely insensitive to any kind of positive and negative effectors and able to ensure at least 95% monomer conversion under the standard conditions of 1 h reaction time at room temperature.(ABSTRACT TRUNCATED AT 250 WORDS)

On the kinetics of monomer incorporation into polyacrylamide gels, as investigated by capillary zone electrophoresis.

The kinetics of monomer incorporation into a polyacrylamide gel have been studied in a photopolymerization system comprising 100 microM methylene blue in presence of a red-ox system, 1 mM sodium toluenesulfinate (reducer) and 50 microM diphenyliodonium chloride (oxidizer). A precise assessment of gel point (pc) was obtained in a droplet chamber, in which argon was gently bubbled with a fused silica capillary into the reaction mixture. At pc, 50% (+/- 3) acrylamide was incorporated into the matrix, vs. 80% (+/- 4) N,N'-methylenebisacrylamide. This incorporation level remained the same when polymerized in the 2-36 degrees C temperature range. Incorporation continued almost linearly for acrylamide up to 80% conversion. The reaction was continued up to 55 min (at 2 degrees C), at which point bisacrylamide had been essentially consumed (> 99.5% incorporation) and acrylamide had reacted (95%). At 2 degrees C, after gelation, the gel became progressively turbid (the Tyndall effect plateauing at 50 min), but it remained fully transparent if, at the gel point, reaction was continued at 50 degrees C. The consumption of the pendant double bonds of Bis followed the progression of turbidity. It is concluded that, by gelation at 2 degrees C, the nascent chains form clusters held together by hydrogen bonds (melting point at 28 degrees C); such clusters are subsequently "frozen" in the three-dimensional space as the pendant double bonds in the chains react progressively. Such turbid matrices are more porous and less elastic than when the gel is polymerized at 50 degrees C. This process is similar to the "lateral aggregation" occurring when gels are formed in presence of a polymer in solution (e.g. 10 KDa polyethylene glycol; Righetti et al., Electrophoresis 1992, 13, 587-594).

On the pH dependence of polymerization efficiency, as investigated by capillary zone electrophoresis.

The conversion efficiency of 5 different catalyst systems for polyacrylamide gel polymerization has been assessed as a function of pH of the gelling solution, in the pH 4.0-10.0 range. The system of persulfate-N,N,N',N'-tetramethylethylene-diamine (TEMED) gives optimal incorporation of monomers only in the pH 7.0-10.0 range; at progressively acidic pH values, the conversion drops markedly until, at pH 4.0, no gelation occurs. In the system of riboflavin-TEMED, the opposite behavior is observed: good conversions in the pH 4.0-7.0 interval (with a peak at pH 6.2), followed by progressive worsening at alkaline pH values until, at pH 10.0, no gelation occurs. An excellent catalyst system appears to be the complex comprising methylene blue, toluene-4-sulfinic acid and diphenyliodonium chloride, which is able to guarantee > 95% conversion in the pH 4.0-8.0 range. Lower efficiencies are obtained at pH 9.0 and 10.0 (down to 81% conversion) but gelation still occurs in the entire pH range. Two other systems (ascorbic acid, ferrous sulfate and hydrogen peroxide in one case, and persulfate, TEMED and hydrosulfite, in another case) have been investigated. The former has some efficiency only at pH 4 (81% conversion), with a rapid drop as pH increases (only 48% incorporation at pH 6). The latter exhibits the opposite behavior: 70% efficiency at pH 4.0, increasing to 92% conversion at pH 6.0.

Photopolymerization of polyacrylamide gels with methylene blue.

Photopolymerization of polyacrylamide gels in the presence of methylene blue (100 microM) and a redox couple (1 mM sodium toluenesulfinate, a reducer, and 50 microM diphenyliodonium chloride, an oxidizer) has been investigated. The gel point, i.e. the time needed for onset of gelation upon illumination, has been found to lengthen progressively at lower temperatures and at lower light intensities. If the three catalysts are progressively diluted, the gel point does not vary for a threefold dilution, but gelation is greatly hampered below a 1:5 dilution of the three effectors. Photobleaching has been assessed as a function of liquid layer thickness (from 0.5 to 2 mm), of a progressive dye dilution (down to a fourfold dilution) and as a function of temperature. A maximum of elastic modulus is located in correspondence to a minimum of permeability (both situated at 5% cross-linker). It is found that methylene blue-activated polymerization produces polyacrylamide gels with elastic properties which are higher than in persulfate-activated gels, so far the most popular matrices for electrokinetic separations. Due to the ease of preparation, the full control of all experimental parameters, and the lack of oxidizing power of this catalyst system (as opposed to the strong oxidation power of persulfate catalysis), methylene blue catalysis is advocated as a valid alternative to other redox systems.

'Laterally aggregated' polyacrylamide gels for electrophoresis.

A new method is described for producing highly porous polyacrylamide matrices: polymerization in presence of a preformed hydrophilic polymer. If a standard mixture of monomers (e.g., 5%T, 4%C) is polymerized in presence of, e.g., polyethylene glycol (PEG) 10 kDa, lateral chain aggregation occurs, with formation of large pore sizes. In PEG 10 kDa, the transition from a small- to a large-pore gel is clearly apparent at 0.5% PEG addition and reaches a plateau already at 2.5% PEG. Even with shorter PEG fragments (6.2 and 1 kDa) this transition occurs, but with progressively larger amounts of PEG in solution (up to 25% for the 1 kDa species). Other polymers such as hydroxymethyl cellulose (1000 kDa) and polyvinyl-pyrrolidone (360 kDa and 25 kDa) are also able to elicit this phenomenon. It appears that lateral chain aggregation (before the cross-linking event) is induced via intra-chain hydrogen bonding, since urea and temperature strongly inhibit it, whereas tetramethylurea (an agent quenching hydrophobic interactions) does not hamper it. By scanning electron microscope, it is found that the maximum pore size obtained in a 5%T, 4%C gel in presence of 2.5% PEG 10 kDA is of the order of 0.5 micron, whereas the same 5%T, 4%C control gel would have an average pore diameter of 5 nm. Thus, an increment of pore size of about 2 orders of magnitude is obtained: in these new matrices, a 21000 bp DNA fragment exhibits a much greater migration than in a control gel in which the sample is entrapped at the application site.