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Two constitutional structural schemes are proposed attempting to unify and rationalize a series of focused NMR and chromatographic determinations aimed at providing an integrated picture for the structure of softwood kraft lignin. The complexity of native softwood lignin when coupled with the complexity of the kraft pulping process is known to lead to a rather heterogeneous material that has eluted us to date. The present work embarks at applying state-of-the-art quantitative 1D and 2D NMR methods on carefully isolated softwood kraft lignin samples and fractions. The accumulated data, when coupled with size exclusion chromatography, mass spectrometric analyses and literature accounts that pertain to the chemistry of kraft pulping, provide the following picture for softwood kraft lignin. Softwood kraft lignin is composed of two distinct fractions that can be separated by using anhydrous acetone. The acetone insoluble fraction is a somewhat branched polymeric material that still contains a variety of native wood lignin bonding patterns, albeit in significantly reduced abundance, as well as new structures induced during the process. The acetone soluble fraction is a significantly more branched and less polymeric material with an abundance of chemical structures that may be created when oligomeric phenols react under kraft pulping conditions. To account for the presence of the various moieties in these two fractions, kraft pulping fragmentation and repolymerization chemistries are extensively invoked, including radical processes initiated by sulfur.

Throughout the long period of human history, mushrooms played a very important role in some societies, where they were used as a highly appreciated food and medicine and as a psychoactive substances in some religious rites. As extremely healthy and nutritious foodstuffs, low in calories, fats, and essential fatty acids, and on the other hand high in vegetable proteins, vitamins, and minerals, mushrooms are recommended and desirable in our daily diet. Most people prefer to eat them fresh due to much more attractive flavor and texture, which rarely can be compared to that of processed mushrooms. However, mushrooms are highly perishable foods, subject to extremely rapid microbiologic and biochemical changes. In an effort to extend their shelf life and thus make them available to more people, over a longer period, several preserving methods have been developed.

A novel and reproducible method is described for accurately determining the molecular weight distribution by size exclusion chromatography (SEC) of whole lignocellulosic materials. This approach offers the opportunity to compare the molecular weight distributions of intact milled woods and its component fractions, lignins and holocelluloses, all from the same source, thus highlighting the potential of the technique and the contributions of the individual components to the chromatogram. The method is based on the dissolution of the ball-milled samples in the ionic liquid 1-allyl-3- methylimidazolium chloride ([amim]Cl). Under these homogeneous ionic liquid media, a derivatization reaction was performed with benzoyl chloride in the presence of pyridine. The thoroughly benzoylated wood with its associated carbohydrate and lignin components was found to be completely soluble in the THF SEC eluent with marked UV detector sensitivity. This methodology, when applied to the individually isolated holocellulose and lignin (enzymatic mild acidolysis lignin; EMAL) materials from Norway spruce (Eucalyptus grandis) wood and corn stover, offered a better understanding as to the possible ways the lignin and the carbohydrates may interact within these three different species. Finally, the applicability of the methodology is shown for a series of pure cellulosic samples under intense mechanical defibration conditions, offering a visualization of the molecular weight distribution changes induced during the production of nanofibrillated cellulose. © 2011 American Chemical Society.

In this paper, we use our quantitative 31P NMR spin trapping methods, already developed for simple oxygen- and carbon-centered radicals, to understand the radical intermediates generated by enzymatic systems and more specifically lipoxygenases. Our methodology rests on the fact that free radicals react with the nitroxide phosphorus compound, 5-diisopropoxy-phosphoryl-5- methyl-1-pyrroline-N-oxide (DIPPMPO), to form stable radical adducts, which are suitably detected and accurately quantified using 31P NMR in the presence of a phosphorus containing internal standard. This system was thus applied to better understand the mechanism of enzymatic oxidation of linoleic acid by soybean lipoxygenases-1 (LOX). The total amount of radicals trapped by DIPPMPO was detected by 31P NMR at different experimental conditions. In particular the effect of dioxygen concentration on the amount of radicals being trapped was studied. At low dioxygen concentration, a huge increase of radicals trapped was observed with respect to the amount of radicals being trapped at normal dioxygen concentrations. © 2011 Published by Elsevier Ltd.

5-Diisopropyloxy-phosphoryl-5-methyl-1-pyrroline-N-oxide (DIPPMPO) was used to trap a variety of free radicals and the stable compounds generated by the natural decomposition of the initially formed spin adducts were characterized by (31)P nuclear magnetic resonance (NMR) and mass spectrometry. Initially, the starting spin trap DIPPMPO was completely characterized using GC-MS and its fragmentation pathway was studied in detail. Then, DIPPMPO was used to trap an oxygen-centered free radical (the hydroxyl radical *OH) and two carbon- centered free radicals (methyl *CH(3) and 1-phenyl-ethanol-1-yl *CCH(3)(OH)Ph radicals). The (31)P NMR signals were thus assigned and the structures of adducts were studied and confirmed by mass spectrometry. Overall, the fragmentation pathways of the radical adducts proceed mainly via the loss of the diisopropyloxy(oxido)phosphoranyl radical. For the specific case of trapping *OH radicals, it is possible to visualize the rearrangement of the nitroxide radical adduct to its nitrone form as invoked in the literature. This spin trapping technique, coupled with (31)P NMR and MS, provides a tool for the identification of short-lived and low molecular weight free radicals present in a variety of processes.

Recent work in our laboratory has allowed the development of quantitative P-31 NMR spin trapping techniques. These methods have been demonstrated to be effective tools for the detection and absolute quantification of many oxygen- and carbon-centered free radical species. Our methods rest on the fact that a free radical reacts with the nitroxide phosphorus compound, 5-diisopropoxy-phosphoryl-5-methyl-1-pyrroline-N-oxide (DIPPMPO), to form stable radical adducts, which are suitably detected and accurately quantified using P-31 NMR in the presence of phosphorus containing internal standards. This system was applied for the detection of phenoxy radicals, as an alternative to traditional EPR techniques. More specifically, the phenoxy radicals were produced via the oxidation of different phenols by K3Fe(CN)(6). The P-31 NMR signals for the radical adducts of phenoxy radicals (PhO center dot) were assigned and found to be located at 25.2 ppm. Subsequently, this spin trapping system was applied to the oxidation of various phenols in the presence of peroxidases and 1-hydroxybenzotriazole (HBT) as a mediator: the 2,4,6-trichlorophenol and 2,4,6-tri-tert-butylphenol were oxidized and only phenoxy radical adducts were detected, whereas during the oxidation of 2,4-dimethylphenol and isoeugenol, other adducts were detected and related to radical delocalization. These preliminary efforts demonstrate the efficacy of our methodologies, so that a variety of radical species can now be readily detected and quantified using quantitative P-31 NMR spin trapping techniques

Corn stover, Norway spruce, and Eucalyptus grandis were pulverized to different degrees. These samples were subjected to quantitative analyses, upon the basis of predissolution into the imidazolium chloride-based ionic liquids [amim]Cl and [bnmim]Cl followed by labeling of hydroxyl groups as phosphite esters and quantitative 31P NMR analysis. Analysis of different pulverization degrees provided semiempirical data to chart the solubility of Norway spruce in these ionic liquids. Further method refinment afforded an optimized method of analysis of the lignin phenolic functionalities, without prior isolation of the lignin from the fiber. The lignin in these samples was further enriched using cellulase and acidolysis treatments, allowing for comparison with the fibrous samples. Analysis of all samples charts the polymerized-monomer availability for each stage of the treatment. Conditions required for adequate signal-to-noise ratios in the 31P NMR analysis were established with a notable improvement observed upon the lignin enrichment steps