Fundamental Insights on the Physical and Chemical Properties of Organosolv Lignin from Norway Spruce Bark.

The interest in the bark and the attempt to add value to its utilization have increased over the last decade. By applying an integrated bark biorefinery approach, it is possible to investigate the recovery of compounds that can be used to develop green and sustainable alternatives to fossil-based materials. In this work, the focus is on extracting Norway spruce (Picea abies) bark lignin via organosolv extraction. Following the removal of the extractives and the subcritical water extraction to remove the polysaccharides, a novel cyclic organosolv extraction procedure was applied, which enabled the recovery of lignin with high quality and preserved structure. Main indicators for low degradation and preservation of the lignin structure were a high ß-O-4' content and low amounts of condensed structures. Furthermore, high purity and low polydispersity of the lignin were observed. Thus, the obtained lignin exhibits high potential for use in the direct development of polymer precursors and other bio-based materials. During the extraction sequence, around 70% of the bark was extracted. Besides the lignin, the extractives as well as pectic polysaccharides and hemicelluloses were recovered with only minor degradation, which could potentially be used for the production of biofuel or other high-value products such as emulsifiers or adhesives.

A modified ionization difference UV-vis method for fast quantitation of guaiacyl-type phenolic hydroxyl groups in lignin.

An ionization difference UV-Vis method (Δε-spectrum method) is the most potentially simple method for fast quantitation of phenolic hydroxyl groups (ph-OH) in lignin. However, the underestimated results were calculated from the conventional Δε-spectrum method using one- or two-point wavelengths measurement. In this study, a modified Δε-spectrum method using multi-point wavelengths measurement was developed and the negative absorbance was also considered. Four main typical lignin models, e.g. vanilla alcohol, 5-5 biphenyl, stilbenoid and vanillin, were applied as the guaiacyl-type (G-type) phenolic models for the determination of ph-OH by the modified Δε-spectrum method. The 2-methoxyethanol/water/acetic acid = 8/2/0.2 (V/V/V) was used as the acidic solvent system and the 2-methoxyethanol/0.2 M NaOH solution = 1/9 (V/V) was used as the alkaline solvent system. The ph-OH contents in the spruce milled wood lignin (SMWL) and the spruce Kraft lignin (SKL) were respectively quantified by the modified Δε-spectrum method as 1.078 and 4.348 mmol/g, which were comparable to the counterparts determined by 31P Nuclear Magnetic Resonance Spectroscopy (31P NMR). The results revealed that the modified Δε-spectrum method can provide more accurate and reliable results compared to the conventional method.

Macromolecular Model of the Pectic Polysaccharides Isolated from the Bark of Norway Spruce (Picea abies).

The bark of Norway spruce (Picea abies) contains up to 13% pectins that can be extracted by pressurized hot water, which constitute a valuable renewable resource in second-generation lignocellulosic biorefineries. This article proposes, for the first time, structural molecular models for the pectins present in spruce bark. Pectin fractions of tailored molar masses were obtained by fractionation of the pressurized hot water extract of the inner bark using preparative size-exclusion chromatography. The monosaccharide composition, average molar mass distribution, and the glycosidic linkage patterns were analyzed for each fraction. The pectin fraction with high molecular weight (Mw of 59,000 Da) contained a highly branched RG-I domain, which accounted for 80% of the fraction and was mainly substituted with arabinan and arabinogalactan (type I and II) side chains. On the other hand, the fractions with lower molar masses (Mw = 15,000 and 9000 Da) were enriched with linear homogalacturonan domains, and also branched arabinan populations. The integration of the analytical information from the macromolecular size distributions, domain composition, and branch lengths of each pectin fraction, results in a comprehensive understanding of the macromolecular architecture of the pectins extracted from the bark of Norway spruce. This paves the way for the valorization of spruce bark pectic polymers in targeted applications based on their distinct polymeric structures and properties.

Hydrophobization of cellulose oxalate using oleic acid in a catalyst-free esterification suitable for preparing reinforcement in polymeric composites.

It is common practice to use cellulose as reinforcement and fatty acid as compatibilizer in the preparation of polymeric composites. However, the used catalysts (e.g., pyridine) are usually toxic and should be avoided. In this study, a new type of microcellulose - cellulose oxalate (COX) was chosen as reinforcement to be reacted with oleic acid to prepare hydrophobic fillers in a catalyst-free esterification for different times. For comparison, microcrystalline cellulose (MCC) was also selected. The success of esterification of COX and oleic acid was confirmed but little esterification occurred when MCC was used. After reacting COX with oleic acid for 18 and 48 h, the products showed stable water contact angles of about 130°. Composites of polypropylene with COX or MCC were prepared. Tensile tests showed that for a given reaction time, the COX-based composites exhibited higher values of both Young's modulus and tensile strength than those of MCC-based composites.

Bactericidal surfaces prepared by femtosecond laser patterning and layer-by-layer polyelectrolyte coating.

Antimicrobial surfaces are important in medical, clinical, and industrial applications, where bacterial infection and biofouling may constitute a serious threat to human health. Conventional approaches against bacteria involve coating the surface with antibiotics, cytotoxic polymers, or metal particles. However, these types of functionalization have a limited lifetime and pose concerns in terms of leaching and degradation of the coating. Thus, there is a great interest in developing long-lasting and non-leaching bactericidal surfaces. To obtain a bactericidal surface, we combine micro and nanoscale patterning of borosilicate glass surfaces by ultrashort pulsed laser irradiation and a non-leaching layer-by-layer polyelectrolyte modification of the surface. The combination of surface structure and surface charge results in an enhanced bactericidal effect against both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria. The laser patterning and the layer-by-layer modification are environmentally friendly processes that are applicable to a wide variety of materials, which makes this method uniquely suited for fundamental studies of bacteria-surface interactions and paves the way for its applications in a variety of fields, such as in hygiene products and medical devices.

Influence of Cellulose Charge on Bacteria Adhesion and Viability to PVAm/CNF/PVAm-Modified Cellulose Model Surfaces.

A contact-active antibacterial approach based on the physical adsorption of a cationic polyelectrolyte onto the surface of a cellulose material is today regarded as an environment-friendly way of creating antibacterial surfaces and materials. In this approach, the electrostatic charge of the treated surfaces is considered to be an important factor for the level of bacteria adsorption and deactivation/killing of the bacteria. In order to clarify the influence of surface charge density of the cellulose on bacteria adsorption as well as on their viability, bacteria were adsorbed onto cellulose model surfaces, which were modified by physically adsorbed cationic polyelectrolytes to create surfaces with different positive charge densities. The surface charge was altered by the layer-by-layer (LbL) assembly of cationic polyvinylamine (PVAm)/anionic cellulose nanofibril/PVAm onto the initially differently charged cellulose model surfaces. After exposing the LbL-treated surfaces to Escherichia coli in aqueous media, a positive correlation was found between the adsorption of bacteria as well as the ratio of nonviable/viable bacteria and the surface charge of the LbL-modified cellulose. By careful colloidal probe atomic force microscopy measurements, it was estimated, due to the difference in surface charges, that interaction forces at least 50 nN between the treated surfaces and a bacterium could be achieved for the surfaces with the highest surface charge, and it is suggested that these considerable interaction forces are sufficient to disrupt the bacterial cell wall and hence kill the bacteria.

Preparation of cellulose nanomaterials via cellulose oxalates.

Nanocellulose prepared from cellulose oxalate has been discussed as an alternative to other methods to prepare cellulose nanofibrils or crystals. The current work describes the use of a bulk reaction between pulp and oxalic acid dihydrate to prepare cellulose oxalate followed by homogenization to produce nanocellulose. The prepared nanocellulose is on average 350 nm long and 3-4 nm wide, with particles of size and shape similar to both cellulose nanofibrils and cellulose nanocrystals. Films prepared from this nanocellulose have a maximum tensile stress of 140-200 MPa, strain at break between 3% and 5%, and oxygen permeability in the range of 0.3-0.5 cm3 µm m-2 day-1 kPa-1 at 50% relative humidity. The presented results illustrate that cellulose oxalates may be a low-cost method to prepare nanocellulose with properties reminiscent of those of both cellulose nanofibrils and cellulose nanocrystals, which may open up new application areas for cellulose nanomaterials.

Changes in chemical structures of wheat straw auto-hydrolysis lignin by 3-hydroxyanthranilic acid as a laccase mediator.

3-Hydroxyanthranilic acid (3-HAA), as a potential natural laccase mediator, was shown to mediate the oxidation of non-phenolic lignin subunits. The problem of cost and toxicity of artificial mediators could be solved to some extent by a further study about the detailed changes of lignin chemistry structures in laccase 3-HAA system (LHS). In this work, wheat straw auto-hydrolysis lignin (AL) was prepared. Oxidations of AL by LHS and laccase 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) system were then investigated, respectively. Various structural changes of AL during the oxidation were characterized by different methods including phenolic hydroxyl group determination, nitrobenzene oxidation, ozonation, gel permeation chromatography, ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy and two-dimensional nuclear magnetic resonance (2D NMR) spectroscopy. The changes in AL chemical structures were found in LHS, including unit removal, bond cleavage and biopolymerization. Compared to laccase ABTS system, a selective removal of guaiacyl-type lignin in LHS was observed, based on the results of nitrobenzene oxidation and 2D NMR analysis. The selective removal of guaiacyl-type lignin was due to improved aromatic ring cleavage and weaken lignin biopolymerization. The selectivity of guaiacyl-type lignin removal in LHS plays an important role, especially for improving bioconversion efficiency of laccase for guaiacyl-rich lignocellulosic biomass.

In Situ Preparation of Silver Nanoparticles in Paper by Reduction with Alkaline Glucose Solutions.

Percolation of contaminated water through paper sheets containing silver nanoparticles is a promising way to provide emergency drinking water. The silver nanoparticles are deposited by the in situ reduction of silver nitrate on the cellulose fibers of an absorbent blotting paper sheet. Sodium borohydride has been used as the reductant but is toxic and expensive. Glucose is a benign alternative but is much less reactive. In this note, we demonstrate an improved way to produce silver nanoparticles in paper sheets by adding sodium hydroxide to the glucose reductant. The silver content of the sheets, measured by diffuse reflectance spectroscopy, was around 2-3 mg of silver per gram of dry paper. This was sufficient to reduce the concentration of a model Escherichia coli suspension after percolation through the sheet.

Water Purification Using Functionalized Cellulosic Fibers with Nonleaching Bacteria Adsorbing Properties.

Portable purification systems are easy ways to obtain clean drinking water when there is no large-scale water treatment available. In this study, the potential to purify water using bacteria adsorbing cellulosic fibers, functionalized with polyelectrolytes according to the layer-by-layer method, is investigated. The adsorbed polyelectrolytes create a positive charge on the fiber surface that physically attracts and bonds with bacteria. Three types of cellulosic materials have been modified and tested for the bacterial removal capacity in water. The time, material-water ratio and bacterial concentration dependence, as well as the bacterial removal capacity in water from natural sources, have been evaluated. Freely dispersed bacteria adsorbing cellulosic fibers can remove greater than 99.9% of Escherichia coli from nonturbid water, with the most notable reduction occurring within the first hour. A filtering approach using modified cellulosic fibers is desirable for purification of natural water. An initial filtration test showed that polyelectrolyte multilayer modified cellulosic fibers can remove greater than 99% of bacteria from natural water. The bacteria adsorbing cellulosic fibers do not leach any biocides, and it is an environmentally sustainable and cheap option for disposable water purification devices.

Bacterial adhesion to polyvinylamine-modified nanocellulose films.

Cellulose nanofibril (CNF) materials have been widely studied in recent years and are suggested for a wide range of applications, e.g., medical and hygiene products. One property not very well studied is the interaction between bacteria and these materials and how this can be controlled. The current work studies how bacteria adhere to different CNF materials modified with polyelectrolyte multilayers. The tested materials were TEMPO-oxidized to have different surface charges, periodate-oxidized to vary the water interaction and hot-pressed to alter the surface structure. Then, multilayers were constructed using polyvinylamine (PVAm) and polyacrylic acid. Both the material surface charge and water interaction affect the amount of polymer adsorbed to the surfaces. Increasing the surface charge decreases the adsorption after the first PVAm layer, possibly due to conformational changes. Periodate-oxidized and crosslinked films have low initial polymer adsorptions; the decreased swelling prevents polymer diffusion into the CNF micropore structure. Microscopic analysis after incubating the samples with bacterial suspensions show that only the materials with the lowest surface charge enable bacteria to adhere to the surface because, when adsorbing up to 5 layers PVAm/PAA, the increased anionic surface charge appears to decrease the net surface charge. Both the amounts of PVAm and PAA influence the net surface charge and thus the bacterial adhesion. The structure generated by the hot-pressing of the films also strongly increases the number of bacteria adhering to the surfaces. These results indicate that the bacterial adhesion to CNF materials can be tailored using polyelectrolyte multilayers on different CNF substrates.

Contact-active antibacterial aerogels from cellulose nanofibrils.

The use of cellulose aerogels as antibacterial materials has been investigated by applying a contact-active layer-by-layer modification to the aerogel surface. Studying the adsorption of multilayers of polyvinylamine (PVAm) and polyacrylic acid to aerogels comprising crosslinked cellulose nanofibrils and monitoring the subsequent bacterial adhesion revealed that up to 26mgPVAmgaerogel(-1) was adsorbed without noticeably affecting the aerogel structure. The antibacterial effect was tested by measuring the reduction of viable bacteria in solution when the aerogels were present. The results show that >99.9% of the bacteria adhered to the surface of the aerogels. Microscopy further showed adherence of bacteria to the surfaces of the modified aerogels. These results indicate that it is possible to create materials with three-dimensional cellulose structures that adsorb bacteria with very high efficiency utilizing the high specific surface area of the aerogels in combination with their open structure.

Cellulose Nanocrystals from Forest Residues as Reinforcing Agents for Composites: A Study from Macro- to Nano-Dimensions.

This study investigates for the first time the feasibility of extracting cellulose nanocrystals (CNCs) from softwood forestry logging residues (woody chips, branches and pine needles), with an obtained gravimetric yield of over 13%. Compared with the other residues, woody chips rendered a higher yield of bleached cellulosic fibers with higher hemicellulose, pectin and lignin content, longer diameter, and lower crystallinity and thermal stability. The isolation of CNCs from these bleached cellulosic fibers was verified by the removal of most of their amorphous components, the increase in the crystallinity index, and the nano-dimensions of the individual crystals. The differences in the physico-chemical properties of the fibers extracted from the three logging residues resulted in CNCs with specific physico-chemical properties. The potential of using the resulting CNCs as reinforcements in nanocomposites was discussed in terms of aspect ratio, crystallinity and thermal stability.

Bioinspired composites from cross-linked galactoglucomannan and microfibrillated cellulose: Thermal, mechanical and oxygen barrier properties.

In this study, new wood-inspired films were developed from microfibrillated cellulose and galactoglucomannan-lignin networks isolated from chemothermomechanical pulping side streams and cross-linked using laccase enzymes. To the best of our knowledge, this is the first time that cross-linked galactoglucomannan-lignin networks have been used for the potential development of composite films inspired by woody-cell wall formation. Their capability as polymeric matrices was assessed based on thermal, structural, mechanical and oxygen permeability analyses. The addition of different amounts of microfibrillated cellulose as a reinforcing agent and glycerol as a plasticizer on the film performances was evaluated. In general, an increase in microfibrillated cellulose resulted in a film with better thermal, mechanical and oxygen barrier performance. However, the presence of glycerol decreased the thermal stability, stiffness and oxygen barrier properties of the films but improved their elongation. Therefore, depending on the application, the film properties can be tailored by adjusting the amounts of reinforcing agent and plasticizer in the film formulation.

Isolation and characterization of cellulose nanocrystals from spruce bark in a biorefinery perspective.

The present study reports for the first time the isolation of cellulose fibers and cellulose nanocrystals (CNCs) from the bark of Norway spruce. The upgrading of bark cellulose to value-added products, such as CNCs, is part of the "bark biorefinery" concept. The removal of non-cellulosic constituents was monitored throughout the isolation process by detailed chemical composition analyses. The morphological investigation of the CNCs was performed using AFM and showed the presence of nanocrystals with an average length of 175.3 nm and a diameter of 2.8 nm, giving an aspect ratio of around 63. X-ray diffraction (XRD) analyses showed that the crystallinity index increased with successive treatments to reach a final value greater than 80% for CNCs. The thermal degradation of the isolated bark CNCs started at 190 °C. Spruce bark appeared to be a new promising industrial source of cellulose fibers and CNCs.

Thermal degradation behavior and kinetic analysis of spruce glucomannan and its methylated derivatives.

The thermal degradation behavior and kinetics of spruce glucomannan (SGM) and its methylated derivatives were investigated using thermogravimetric analysis to characterize its temperature-dependent changes for use in specific applications. The results were compared with those obtained for commercial konjac glucomannan (KGM). The SGM and the KGM exhibited two overlapping peaks from 200 to 375°C, which correspond to the intensive devolatilization of more than 59% of the total weight. Differences in the pyrolysis-product distributions and thermal stabilities appeared as a result of the different chemical compositions and molecular weights of the two GMs. The Friedman and Flynn-Wall-Ozawa isoconversional methods and the Coats-Redfern were adopted to determine the kinetic triplet of the intensive devolatilization region. Both GMs can be modeled using a complex mechanism that involves both a Dn-type and an Fn-type reaction. The comparative study of partially methylated GM indicated higher homogeneity and thermal resistance for the material with the higher degree of substitution.

Hot-water extracts from the inner bark of Norway spruce with immunomodulating activities.

The inner bark of Norway spruce (Picea abies) was sequentially extracted with hot water at 100°C, 140°C and 160°C. The hot-water extracts (IB 100°C, IB 140°C and IB 160°C) contained pectic polysaccharides and showed immunostimulating activities. Structural analyses of their carbohydrate content, including glycosidic linkage analyses, revealed the presence of pectins with a large rhamnogalacturonan RG-I domain ramified with highly-branched arabinans. IB 100°C also contained a large amount of terminal glucosyl residues, indicating the presence of highly substituted polymers. IB 160°C was mainly composed of starch. The hot-water extracts were tested for two biological activities, namely complement fixation and macrophage stimulation. IB 100°C exhibited the highest complement fixation activity, with a 1.7-times higher ICH50 than the control pectin, while IB 140°C and IB 160°C gave similar ICH50 values as the control. Macrophages were stimulated by IB 100°C and IB 140°C in a dose-dependent manner, but not by IB 160°C. IB 100°C presented the highest activity toward macrophages, comparable to the control pectin.

Bacterial-growth inhibiting properties of multilayers formed with modified polyvinylamine.

New methods are needed to fight antibiotic-resistant bacteria. One alternative that has been proposed is non-leaching, permanently antibacterial surfaces. In this study, we test multilayers formed with antibacterial cationic polyvinylamine (PVAm) and polyacrylic acid (PAA) in a growth-inhibition assay. Both hydrophobically modified and native PVAm were investigated. Multilayers did reduce the bacterial growth, as compared to single layers. However, the sampling time in the assay was critical, as the treated surface area is a capacity-limiting factor. After 2h incubation, a maximal growth inhibition of more than 99% was achieved with multilayers. In contrast, after 8h we observed a maximal growth-inhibition of 40%. At longer incubation times, the surface becomes saturated, which explains the observed time-dependent effectiveness. The polymers giving multilayers with the strongest growth-inhibiting properties were native PVAm and PVAm modified with C(8), which also were the polymers with highest charge density. We therefore conclude that this effect is mainly an electrostatically driven process. Viability staining using a fluorescent stain showed a high viability rate of the adhered bacteria. The multilayers are therefore more bacteriostatic than antibacterial.

Combination of alkaline and enzymatic treatments as a process for upgrading sisal paper-grade pulp to dissolving-grade pulp.

A sequence of treatments consisting of an initial xylanase treatment followed by cold alkaline extraction and a final endoglucanase treatment was investigated as a process for upgrading non-wood paper-grade pulps to dissolving-grade pulps for viscose production. Five commercial dried bleached non-wood soda/AQ paper pulps, from flax, hemp, sisal, abaca, and jute, were studied for this purpose. Commercial dried bleached eucalyptus dissolving pulp was used as reference sample. Sisal pulp showed the highest improvement in Fock's reactivity, reaching levels nearly as high or even higher than that of eucalyptus dissolving pulp (65%), and a low hemicellulose content (3-4%) when was subjected to this sequence of treatments. The viscosity, however, decreased considerably. A uniform and narrow molecular weight distribution was observed by size exclusion chromatography. (13)C nuclear magnetic resonance spectroscopy and Raman microspectroscopy revealed that the cellulose structure consisted of cellulose I.

Interactions of hydrophobically modified polyvinylamines: adsorption behavior at charged surfaces and the formation of polyelectrolyte multilayers with polyacrylic acid.

The structure and adsorption behaviors of two types of hydrophobically modified polyvinylamines (PVAm) containing substituents of hexyl and octyl chains were compared to a native polyvinylamine sample. The conformation of dissolved polyvinylamines was studied in aqueous salt solutions using dynamic light scattering. Modified PVAm showed hydrodynamic diameters similar to native PVAm, which indicated that all PVAm polymers were present as single molecules in solution. The adsorption of the polyvinylamines, both native and hydrophobically modified, from aqueous solution onto negatively charged silica surfaces was studied in situ by reflectometry and quartz crystal microgravimetry with dissipation. Polyelectrolyte multilayers (PEM) with up to nine individual layers were formed together with poly(acrylic acid). Obtained PEM structures were rigid and showed high adsorbed amounts combined with low dissipation, with similar results for both the modified and unmodified PVAm. This suggests that electrostatics dominated the PEM formation. At lower salt concentrations, the hydrophobically modified PVAm produced multilayers with low water contents, indicating that secondary interactions induced by the hydrophobic constituents can also have a significant influence on the properties of the formed layers. The surface structure of PEMs with nine individual layers was imaged in dry state using atomic force microscopy in a dynamic mode. Modified PVAm was found to induce a different structure of the PEM at 100 mM, with larger aggregates compared to those of native PVAm. From these results, it is proposed that modified PVAm can induce aggregation within the PEM, whereas PVAm remains as single molecules in solution.