Optimization of the extraction of galactoglucomannans from Pinus halepensis

The effectiveness of pressurized hot-water extraction conditions for obtaining galactoglucomannans (GGMs) from Pinus halepensis suitable for applications like coatings and films packaging was investigated. For this purpose, high molar masses with high yields are required, presenting a serious challenge for hot-water extraction processes. The extraction of GGMs was carried out in an accelerated solvent extractor (ASE) and the isolation was performed by precipitation in ethanol. Three temperatures in the range 160-180 degrees C and five extraction times 5-90 min were tested in order to optimize extraction parameters of GGMs, avoiding thermal and chemical degradation in hotwater. Total dissolved solids (TDS) were determined gravimetrically after freeze-drying and weight average molar masses (Mw) were determined by high-performance size exclusion chromatography (HPSEC). Total non-cellulosic carbohydrates were determined by gas chromatography (GC) after acid methanolysis. Free monomers were additionally analyzed by GC. Lignin in water extracts was measured by an ultraviolet (UV) method. Acetic acid was determined after alkaline hydrolysis of acetyl groups and analyzed by HPSEC. The main parameters influencing the extraction processes of the GGMs, namely, extraction time and temperature were studied. Optimal extraction parameters of GGMs were identified at 170 degrees C and 20 min extraction time, with average Mw of extracted fraction of 7 kDa leading to a GGM yield of approximately 56 mg g(o.d.m)(-1) corresponding to 6% on dry wood basis.This work was part of the doctoral research activities of ((insert author name/s)) at the Johan Gadolin Process Chemistry Centre (PCC) at Abo Akademi University, Finland. We thank The Algerian Ministry of Higher Education and Scientific Research for funding support within "Le Projet National Exceptionnel (P.N.E)'

Localization of (+)-Catechin in Picea abies Phloem : Responses to Wounding and Fungal Inoculation

To understand the positional and temporal defense mechanisms of coniferous tree bark at the tissue and cellular levels, the phloem topochemistry and structural properties were examined after artificially induced bark defense reactions. Wounding and fungal inoculation withEndoconidiophora polonicaof spruce bark were carried out, and phloem tissues were frequently collected to follow the temporal and spatial progress of chemical and structural responses. The changes in (+)-catechin, (-)-epicatechin, stilbene glucoside, and resin acid distribution, and accumulation patterns within the phloem, were mapped using time-of-flight secondary ion mass spectrometry (cryo-ToF-SIMS), alongside detailed structural (LM, TEM, SEM) and quantitative chemical microanalyses of the tissues. Our results show that axial phloem parenchyma cells of Norway spruce contain (+)-catechins, the amount of which locally increases in response to fungal inoculation. The preformed, constitutive distribution and accumulation patterns of (+)-catechins closely follow those of stilbene glucosides. Phloem phenolics are not translocated but form a layered defense barrier with oleoresin compounds in response to pathogen attack. Our results suggest that axial phloem parenchyma cells are the primary location for (+)-catechin storage and synthesis in Norway spruce phloem. Chemical mapping of bark defensive metabolites by cryo-ToF-SIMS, in addition to structural and chemical microanalyses of the defense reactions, can provide novel information on the local amplitudes and localizations of chemical and structural defense mechanisms and pathogen-host interactions of trees.Peer reviewe

Pilot Study of Sap Properties of Norway Spruce (Picea abies (L.) Karst.) Trees Used and Not Used for Sap- Feeding by Three-Toed Woodpeckers (Picoides tridactylus)

The ecophysiology of the Norway spruce (Picea abies (L.) Karst.) trees that were used by three-toed woodpeckers (TTW) (Picoides tridactylus) for their sap-feeding activities were investigated. The pilot study was conducted in southern Finland (61°15′ N, 25°00′ E). During April–June 2015, three different tree categories of Norway spruce were selected for monitoring: trees that were frequently used by the TTWs for phloem sap-feeding for several previous years; trees that were only recently used by TTWs for sap-feeding; and control trees that were not used at all for sap-feeding. Phloem sap and phloem tissue samples were frequently extracted from tree trunks and analyzed for the content and composition of nonstructural carbohydrates, phloem sap osmolality, solute, and water content, as well as for the content and composition of secondary metabolites typical for defense reactions in the phloem. Simple crown characteristics were also measured, including tree height, diameter at breast height, and their ratio (slenderness index). According to our results, the TTWs preferred Norway spruce trees that showed advanced spring phenology to feed on, as evidenced especially by the lower ratio of raffinose (typically high during the winter months) to total soluble sugars of phloem tissues as compared to non-used control trees. The lower slenderness index of the trees chosen by the TTWs indicates low canopy competition pressure with good access to light (i.e., the sun heats the trunks well in spring). There were no differences in the phloem osmolality or solute content between the used or unused control trees. The trees used by the TTWs had significantly higher concentrations of antioxidant phenolic (+)-catechins and stilbene glycosides in phloem tissue, and the stilbene content was also higher in the extracted sap. The phenolics content of the phloem tissue had a clear seasonal trend, being the highest in the early spring and lower towards the onset of the cambial growth processes. The phloem sap is rich in antioxidants and soluble sugars that are potentially beneficial for the TTWs, but more quantitative research is needed to explore the importance of the sap properties to TTWs

Lignans in Knotwood of Norway Spruce : Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy

Lignans are bioactive compounds that are especially abundant in the Norway spruce (Picea abiesL. Karst.) knotwood. By combining a variety of chromatographic, spectroscopic and imaging techniques, we were able to quantify, qualify and localise the easily extractable lignans in the xylem tissue. The knotwood samples contained 15 different lignans according to the gas chromatography-mass spectrometry analysis. They comprised 16% of the knotwood dry weight and 82% of the acetone extract. The main lignans were found to be hydroxymatairesinols HMR1 and HMR2. Cryosectioned and resin-embedded ultrathin sections of the knotwood were analysed with scanning transmission X-ray microscopy (STXM). Cryosectioning was found to retain only lignan residues inside the cell lumina. In the resin-embedded samples, lignan was interpreted to be unevenly distributed inside the cell lumina, and partially confined in deposits which were either readily present in the lumina or formed when OsO(4)used in staining reacted with the lignans. Furthermore, the multi-technique characterisation enabled us to obtain information on the chemical composition of the structural components of knotwood. A simple spectral analysis of the STXM data gave consistent results with the gas chromatographic methods about the relative amounts of cell wall components (lignin and polysaccharides). The STXM analysis also indicated that a torus of a bordered pit contained aromatic compounds, possibly lignin.Peer reviewe

Environmentally-compatible alkyd paints stabilized by wood hemicelluloses

Wood biorefining currently involves large-scale industrial processes where a notable portion of raw materials, namely hemicelluloses and lignin, are either lost with the process water, degraded, or burnt for energy. Value added utilization of polymeric hemicelluloses is challenging due to their intermediate molar mass and the presence of other wood components, such as phenolic residues or wood extractives. Oil-in-water (O/W) emulsions represent a diverse and abundant class of applications in which the natural properties of wood hemicelluloses are beneficial. In the current work, we present highly promising new technical alkyd paint emulsion systems stabilized with hardwood glucuronoxylans (GX) and softwood galactoglucomannans (GGM). Samples from three isolation methods and their further fractionation by ethanol precipitation were systematically compared with regard to hemicellulose composition, interfacial activity, and functionality in emulsions. Emulsification of alkyd resins was successful with both GX and GGM obtained by various biorefining strategies. The highest emulsion stability over storage was achieved using crude non-purified GX and GGM fractions, and was correlated with the presence of phenolic compounds and extractives, interfacial activity, and small droplet size. Hardwood GX and softwood GGM are envisioned as natural emulsifiers of alkyd O/W emulsions, which are examples of diverse and abundantly-used technical dispersions. This study can be utilized as a guideline for targeted extraction of hemicelluloses with desired functionality, and as a protocol for developing environmentally-compatible industrial dispersions.Peer reviewe

Fractionation of Lignin with Decreased Heterogeneity: Based on a Detailed Characteristics Study of Sequentially Extracted Softwood Kraft Lignin

Industrial lignin fractionation is attracting increasing interest due to its enormous potential in the development of high value-added materials. However, the widely reported fractionation approaches are primarily focused on the separation of fractions with a low polydispersity index (PDI). In this study, based on the detailed characteristic examination of carefully sequential-extracted softwood Kraft lignin fractions, a novel method to isolate lignin fraction with decreased heterogeneity (LGF-dh), was established in consideration of impurities, elemental composition, molar mass distribution, carbohydrate content, functional hydroxyl content, and the content of lignin-relevant aromatic units. To characterize the mentioned properties, an elemental analyzer, SEC-MALS, GC-MS, GC-FID, Py/GCMS, P-31-NMR, and HSQC-NMR were used to compare the differences of the sequential lignin fractions that were obtained by methyl tert-butyl ether (MTBE), ethyl acetate (EtOAc), ethanol (EtOH), methanol (MeOH), acetone, and dioxane. Moreover, a practical and feasible three-step extraction process was proposed to separate the low heterogeneity lignin fraction from industrial lignin according to the different solubilities of each fraction in the green cosolvent system of EtOH/water, MeOH/water, and acetone/water. Overall, this work presented a comprehensive study on the properties of softwood lignin as well as proposed a feasible and convenient method to reduce the heterogeneity of lignin, which would promote its valorization

HOT-WATER EXTRACTION OF GROUND SPRUCE WOOD OF DIFFERENT PARTICLE SIZE

Hot-water extraction of hemicelluloses, especially galactoglucomannans, from fractions of ground spruce wood with different particle sizes was studied at 170°C with extraction times up to 60 min. Extraction of spruce sapwood, heartwood, and thermomechanical pulp (TMP) was also compared at 160 to 180°C. Static batch extractions were carried out in an accelerated solvent extractor (ASE). The extracted hemicelluloses were characterized by sugar unit analysis and determination of acetyl groups and molar masses. The particle size significantly affected the extraction of ground wood. The total extraction yield, as well as the yields of hemicelluloses and monosaccharides, was the highest from the finest ground wood fraction (< 0.1 mm). The release of acetic acid, average molar mass of extracted hemicelluloses, and end-pH of the extracts were also dependent of the particle size, although to a lower extent. Irrespectively of the ground wood particle size, the yield of hemicelluloses reached a plateau after 40 min extraction at 170°C. The results indicate that extraction of hemicelluloses is limited mainly by the diffusion in the fiber wall, and for coarse wood shives also by the mass transfer in the wood matrix. There were only small differences in the hot-water extraction yields of hemicelluloses from spruce sapwood, heartwood, and TMP, considering both poly- and monosaccharides