Characterization of the lignin-derived products from wood as treated in supercritical water

Sugi (Cryptomeria japonica D. Don) and buna (Fugus crenata Blume) woods were treated with supercritical water (>374°C, >22.1 MPa) and fractionated into a water-soluble portion and a water-insoluble residue. The latter was washed with methanol to be fractionated further into a methanol-soluble portion and a methanol-insoluble residue. Whereas the carbohydrate-derived products were in the water-soluble portion, most of the lignin-derived products were found in the methanol-soluble portion and methanol-insoluble residue. The lignin-derived products in the methanol-soluble portion were shown to have more phenolic hydroxyl groups than lignin in original wood. The alkaline nitrobenzene oxidation analyses, however, exhibited much less oxidation product in the methanol-soluble portion and methanol-insoluble residue. These lines of evidence suggest that the ether linkages of lignin are preferentially cleaved during supercritical water treatment. To simulate the reaction of lignin, a study with lignin model compounds was performed;-O-4-type lignin model compounds were found to be cleaved, whereas biphenyl-type compounds were highly stable during supercritical water treatment. These results clearly indicated that the lignin-derived products, mainly consisting of condensed-type linkages of lignin due to the preferential degradation of the ether linkages of lignin, occurred during supercritical water treatment.This study was presented in part at the 45th lignin symposium, Ehime, Japan, October, 2000; and the 49th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1988     

Characterization of lignin-derived products from Japanese beech wood as treated by two-step semi-flow hot-compressed water

Japanese beech (Fagus crenata) wood was treated by two-step semi-flow hot-compressed water (the first stage: 230 °C/10 MPa/15 min, the second stage: 270 °C/10 MPa/15 min), and produced lignin-derived products in the hot-compressed water-soluble portions at the first and second stages, and the final residue of the second stage was characterized with alkaline nitrobenzene oxidation method and gel permeation chromatographic analysis. As a result, the lignin-derived products at the first stage, where hemicellulose was also decomposed, consisted of lignin-based monomers and dimers and oligomers/polymers in the water-soluble portion. A large part of the oligomers/polymers was, however, recovered as the precipitate during 12 h setting after hot-compressed water treatment. By the analysis of nitrobenzene oxidation products, there were relatively higher contents of ether-type lignin in the precipitate at the first stage than in original beech wood. Since the ether linkages of lignin are more preferentially cleaved by this hot-compressed water, lignin-based polymeric fractions were flowed out from the porous cell walls from which hemicellulose was removed. On the other hand, at the second stage condensed-type lignin remained in the precipitate and residue. Based on these results, decomposition behavior of lignin in Japanese beech wood as treated by the two-step semi-flow hot-compressed water was discussed regarding the topochemistry of lignin structure.     

Effect of ozone treatment of wood on its liquefaction

The effects of ozone treatment were investigated to improve the process of liquefaction of wood with polyhydric alcohol solvents. The liquefied wood having a high wood to polyhydric alcohol ratio (W/P ratio) could be prepared by using the wood treated with ozone in the liquid phase. The liquefied wood with a W/P ratio of 2 : 1 had enough fluidity to act as a raw material for chemical products. To get some information about the effects of ozone treatment toward the wood components, cellulose powder and steamed lignin were treated with ozone and liquefied. In particular, ozone treatment in the liquid phase was found to be effective for wood and cellulose powder. On the other hand, steamed lignin self-condensed during liquefaction after treatment with ozone in the liquid phase. Thus, ozone treatment provided lignin with reactive functional groups, and caused the subsequent condensation reaction. Although lignin was converted to a more condensable structure by ozone treatment, the condensation reaction was found to be suppressed for wood during its liquefaction. The wood liquefied products displayed good solubilities in N,N-dimethyl formamide (DMF) even after treatments of long duration. It was suggested that one of the main effects of ozone treatment toward wood was the decomposition of cellulose.Part of this report was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, April 2003     

Gas chromatographic and mass spectrometric (GC-MS) analysis of lignin-derived products from <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> treated in supercritical water

Sugi (Cryptomeria japonica D. Don) wood was treated with supercritical water (374°C, 22.1MPa), and fractionated into the water-soluble portion, the methanol-soluble portion, and the methanol-insoluble residue. The methanol-soluble portion mainly consisted of the lignin-derived products. To characterize the compounds in the methanol-soluble portion, gel permeation chromatographic (GPC) and gas chromatographic-mass spectrometric (GC-MS) analyses were performed. The GPC analysis indicated that the methanol-soluble portion contained lignin-derived monomeric and dimeric products. GC-MS analysis detected 31 products which were expected to be monomeric compounds, and 18 of these were identified to be guaiacol, methylguaiacol, ethylguaiacol, vinylguaiacol, eugenol, propylguaiacol, vanillin, cis-isoeugenol, homovanillin, trans-isoeugenol, acetoguaiacone, propioguaiacone, guaiacylacetone, 2-methoxy-4-(1-hydroxypropyl)phenol, homovanillic acid, 2-methoxy-4-(prop-1-en-3-one)phenol, coniferyl aldehyde, and ferulic acid. In addition, 22 dimeric products were detected, and 4 of these were believed to be compounds with biphenyl type (5-5), diphenylethane type (-1), stilbene type (-1), and phenylcoumaran type (-5) structures. These results clearly indicated that the methanol-soluble portion included various monomeric and dimeric compounds produced as a result of the cleavage of ether linkages and propyl chains of lignin.     

Chemical characteristics of surfaces of hardwood and softwood deteriorated by weathering

The factors that cause weather-induced deterioration of wood surfaces were determined by chemical and spectroscopic analyses. Albizzia (Paraserianthes falcata Becker.) and sugi (Cryptomeria japonica D. Don) were exposed to two temperate conditions of natural weathering with and without rainfall and to accelerated conditions of artificial weathering coupled with ultraviolet (UV) light irradiation and water flashing. Infrared spectroscopic analysis showed that the oxidative reaction of lignin was observed under all conditions of weathering for both wood species. However, a marked decrease in lignin and hemicellulose content were recognized when albizzia woods were exposed to weathering with water. Lignin content in the softwood sugi did not decrease as much as in albizzia even in the presence of water, but the modification of lignin macromolecules was assumed to be accelerated by water, as seen by electron spin resonance spectroscopy. These results showed that the presence of water promotes the weathering deterioration of wood under UV irradiation.     

Characterization of lignin-derived products from various lignocellulosics as treated by semi-flow hot-compressed water

To elucidate the decomposition behaviors of lignin from different taxonomic groups, five different lignocellulosics were treated with hot-compressed water (230 °C/10 MPa/15 min) to fractionate lignins into water-soluble portions, precipitates, and insoluble residues. The lignin-derived products in each fraction were characterized and compared. The delignification of monocotyledons [nipa palm (Nypa fruticans) frond, rice (Oryza sativa) straw, and corn (Zea mays) cob] was more extensive than that achieved for Japanese cedar (Cryptomeria japonica, gymnosperm) and Japanese beech (Fagus crenata, dicotyledon angiosperm). The water-soluble portions contained lignin monomers like coniferyl alcohol and phenolic acids, while the precipitates contained higher molecular weight lignin with high content of ether-type linkages. Lignin in the insoluble residues was rich in condensed-type structures. In all five lignocellulosics, ether-type linkages were preferentially cleaved, while condensed-type lignin showed resistance to hot-compressed water. In the monocotyledons, lignin–carbohydrate complexes were cleaved and gave lignins that had higher molecular weights than those eluted from the woods. These differences would facilitate the delignification in monocotyledons. Such information provides useful information for efficient utilization of various lignocellulosics.     

Fractionation and characterization of oil palm (Elaeis guineensis) as treated by supercritical water

In order to investigate the potential for efficient utilization of oil palm (Elaeis guineensis), supercritical water treatment (at 380°C and 100 MPa for 8 s) was applied to fractionate extractives-free samples into water-soluble portion and water-insoluble portion. The water-insoluble portion was further fractionated into methanol-soluble portion and methanol-insoluble residue. Samples were prepared from various parts of oil palm, i.e., trunks, fronds, mesocarp fibers, shells, empty fruit bunches, and kernel cake. These fractionated products were then characterized analytically. The water-soluble and methanol-soluble portions were determined to be mainly composed of carbohydrate-derived products and lignin-derived products, respectively. The methanol-insoluble residue was mainly composed of lignin (more than 84 wt%) and the phenolic hydroxyl contents determined by the aminolysis method was higher than for untreated oil palm samples. In addition, an alkaline nitrobenzene oxidation analysis indicated that the methanol-insoluble residue contained fewer oxidation products than untreated samples did. These findings imply that the water-soluble portion could be utilized for organic acid production, whereas the methanol-soluble portion and the insoluble residue could be used for the production of phenolic chemicals.     

Lignification and peroxidase in tension wood ofEucalyptus viminalis seedlings

Seedlings ofEucalyptus viminalis were grown for 50 days with their stems bent so tension wood would form. Every 10 days the lignin content, monomeric composition, and peroxidase activity in the tension wood were compared with those in the lower side (opposite wood) and in vertically grown controls. The lignin content in the developing tension wood started to decrease after 10 days of bending and kept decreasing for 50 days, whereas those in control plants and opposite wood remained almost unchanged. The yields of syringaldehyde from tension wood by nitrobenzene oxidation increased, and consequently the syringyl/ guaiacyl ratio of the lignin was higher in tension wood than in opposite wood and control plants. The peroxidase ionically bound to the cell walls (IPO) catalyzed oxidation of guaiacol and syringaldazine. The syringaldazineoxidizing activity of IPO from tension wood increased, whereas the activities of IPO from opposite wood and control plants did not show any marked change. In tension wood the increase in syringaldazine-oxidizing activity of IPO was consistent with an increase in the syringaldehyde yield. This suggests that IPO contributes to syringyl lignin deposition as other enzymes involved in the monolignol biosynthesis do in tension wood formation.This study was presented at the 50th Annual Meeting of the Japan Wood Research Society, Kyoto, April 2000     

Erythro/threo ratio ofβ-O-4 structures as an important structural characteristic of lignin. I: Improvement of ozonation method for the quantitative analysis of lignin side-chain structure

Ozonation as a quantitative tool to analyze the stereo structures of arylglycerol--aryl ether linkages was examined using wood meal, milled wood lignin, and a lignin model compound, 1-(3,4-dimethoxyphenyl)-2-(2-methoxyphenoxy)-1,3-propanediol (veratrylglycerol--guaiacyl ether, VG). The procedure was improved. When mild postreduction was conducted for ozonation products, the total yield of erythronic and threonic acids from this model compound was 74%, which is 15% higher than the yield without postreduction. A decrease in the recovery of these two acids under prolonged ozonation treatment was successfully suppressed by postreduction. Theerythro/threo ratio of VG determined by the ozonation method with postreduction is in good agreement with the ratio determined by¹H-nuclear magnetic resonance. Excellent reproducibility of the yield was obtained by adopting a procedure that included trimethylsilylation of ammonium salts of ozonation products using a dimethylsulfoxide-hexamethyldisilazane-trimethylchlorosilane mixture and subjecting it to gas chromatography analysis. It was concluded that arylglycerol--aryl ether structures comprise at least 35% of the C₃-C₆ structure in birch wood meal, with anerythro/threo ratio of 2.8.This paper was presented at the 43rd Lignin Symposium, Fuchu, Japan, October 1998; and at the 10th ISWPC, Yokohama, Japan, June 1999     

Treatment of poplar callus with ferulic and sinapic acids I: incorporation and enhancement of lignin biosynthesis

Ferulic acid (FA), tetradeuteroferulic acid (DFA), sinapic acid (SA), or heptadeuterosinapic acid (DSA) was exogenously supplied to poplar (Populus alba L.) callus. Administration of FA or SA increased the lignin content of the callus to about twice that of the control callus. Gas chromatographic analysis of the alkali hydrolysate of the cell wall residue revealed that only a trace amount of SA was bound to the cell wall, and the amount of FA was less than 2% of the total callus lignin. Thioacidolysis of the DFA-treated callus indicated that DFA is effectively converted to both coniferyl and sinapyl alcohols and then incorporated into the corresponding lignin. Incorporation of DSA into syringyl lignin or guaiacyl lignin was not observed, but yields of syringyl lignin thioacidolysis products were markedly increased by DSA treatment of the callus. These results suggest that SA may not be a precursor of sinapyl alcohol and syringyl lignin per se, but it may induce or enhance the biosynthesis of syringyl lignin in poplar callus.     

Leaf quality of selected hedgerow species at two canopy ages in the derived savanna zone of West Africa

Prunings in hedgerow intercropping systems are a potential source of organic matter and their quality is an important characteristic driving decomposition and nutrient release. To determine the potential range of residue characteristics and the impact of canopy age on those, selected characteristics of Leucaena leucocephala, Gliricidia sepium, and Senna siamea hedgerow leaf residues were determined for young (5–10 weeks) and old (39–47 weeks) hedgerow canopies in six sites, representative in terms of soil for the derived savanna zone of West-Africa. The N content of the Leucaena and Senna residues decreased with age (from 4.8 to 3.9% and from 4.1 to 3.0%, respectively, while the N content of the Gliricidia residues remained constant (3.9%). The P content of all species decreased with age (from 0.30 to 0.20%, on average). The lignin content of the Leucaena residues increased, and their polyphenol content decreased with age, while for the Senna residues only an increase in polyphenol content with canopy age was evident. Neither the lignin nor the polyphenol content of the Gliricidia leaf residues was markedly affected by canopy age. For all species, the P and ADF content were positively correlated with the ash and lignin content, respectively. The N content of the Leucaena and Senna residues was positively correlated with their P content while the lignin content of the same species was negatively correlated with the polyphenol content. Because of the low variation in certain residue characteristics between sites, the presented data and equations could potentially be used to reasonably estimate those characteristics if no other information is available. For other characteristics, such as polyphenol contents, measurements are preferable in view of the relatively high variation encountered.This revised version was published online in November 2005 with corrections to the Cover Date.     

Lignin characteristics of bast fiber and core in kenaf, bark and wood of paper mulberry and mulberry

The structural features of bast fiber and core lignins in kenaf (Hibiscus cannabinus), bark and wood lignin of paper mulberry (Broussonetia papyrifera (L.) Vent × Broussonetia kazinoki Sieb.) and mulberry (Morus bombycis) were characterized by alkaline nitrobenzene oxidation, ozonation and methoxyl group determination. Bj?rkman lignins were isolated from bast fiber and core, and bark and wood fractions of the plant samples, and structural characteristics were investigated by ¹H NMR and ¹³C NMR spectroscopies. Kenaf bast fiber gave very high molar ratio of syringaldehyde to vanillin (S/V) of alkaline nitrobenzene oxidation products, while methoxyl content was about the same as that of the core fraction. Results of ¹H NMR and ¹³C NMR of Bj?rkman lignin suggested the presence of aliphatic fragments in lignins isolated from paper mulberry and mulberry bark, but not in kenaf bast fiber. The lower yield of alkaline nitrobenzene oxidation products from bast fiber and bark might be due to the higher content of condensed structure of lignin compared to core fraction. Total yield of erythronic (E) and threonic (T) acids of ozonation products and the molar ratio of erythronic acid to threonic acid (E/T) of the bast fibers and bark were lower than the corresponding core and wood fractions, suggesting that the contents of arylglycerol-β-aryl ether intermonomer linkages in the bast fiber and bark lignin were lower than those of the core and wood fractions. Methoxyl content of bark lignin was lower than the corresponding wood lignin. The methoxyl content of the extract-free kenaf bast fiber was similar to that of the core fraction, while the values of paper mulberry and mulberry bark were about one-half of the corresponding wood fractions, respectively. In bark lignins, the methoxyl contents of Klason lignin and Bj?rkman lignin from bark were lower than those of the extract-free barks. This result suggests that the purity of Klason lignin and Bj?rkman lignins of bark may be rather low.     

Determination of lignin and extractive content of Turkish Pine (<Emphasis Type="Italic">Pinus brutia</Emphasis> Ten.) trees using near infrared spectroscopy and multivariate calibration

Determination of quality parameters such as lignin and extractive content of wood samples by wet chemistry analyses takes a long time. Near-infrared (NIR) spectroscopy coupled with multivariate calibration offers a fast and nondestructive alternative to obtain reliable results. However, due to the complexity of the NIR spectra, some wavelength selection is generally required to improve the predictive ability of multivariate calibration methods. Pinus brutia Ten. is the most growing pine species in Turkey. Its rotation period is around 80 years; the forest products industry has widely accepted the use of Pinus brutia Ten. because of its ability to grow on a wide range of sites and its suitability to produce desirable products. Pinus brutia Ten. is widely used in construction, window door panel, floor covering, etc. Determination of lignin and extractive content of wood provides information to tree breeders on when to cut and how much chemicals are needed for the pulping and bleaching process. In this study, 58 samples of Pinus brutia Ten. trees were collected in Isparta region of Turkey, and their lignin and extractive content were determined with standard reference (TAPPI) methods. Then, the same samples were scanned with near-infrared spectrometer between 1,000 and 2,500 nm in diffuse reflectance mode, and multivariate calibration models were built with genetic inverse least squares method for both lignin and extractive content using the concentration information obtained from wet standard reference method. Overall, standard error of calibration (SEC) and standard error of prediction (SEP) ranged between 0.35% (w/w) and 2.40% (w/w).     

Separation of liquefied product of Salix psammophila by column chromatography and structure analysis of its components

The liquefied product of Salixpsammophila wood was separated by thin-layer chromatography （TLC） and column chromatography, and its structure was identified by nuclear magnetic resonance （NMR） spectra in our study. The separation result indicates that the sample of liquefied S. psammophila contained at least two categories of components. The structure of the main components was guaiacyl C-1, C-2 of the hydroxyphenyl propane, i.e., the aromatic nucleus protons of lignin. Degradation and polycondensation reactions occurred when the S. psammophila wood was liquefied in phenol. Polycondensation reactions occurred among the depolymerization products from cellulose, the aromatic depolymerization products from lignin and the products of the displacement reactions between phenoxide ion and cellulose.     

GC-MS and IR spectroscopic analyses of the lignin-derived products from softwood and hardwood treated in supercritical water

Softwood (Cryptomeria japonica) and hardwood (Fagus crenata) were treated in supercritical water (380°C, 100 MPa) for 8 s. The treated woods were fractionated to the water-soluble portion, methanol-soluble portion, and methanol-insoluble residues. For the methanol-soluble portion, which mainly consisted of lignin-derived products, gel permeation chromatography (GPC) and gas chromatographic-mass spectrometric (GC-MS) analyses were conducted to clarify the molecular weight distribution and to identify the monomeric products, respectively. GPC analysis revealed that the methanol-soluble portion contains monomeric and some oligomeric products. GC-MS analysis identified 19 guaiacyl compounds in the methanol-soluble portion from softwood, and 15 syringyl monomeric compounds in the methanol-soluble portion from hardwood. The structures of identified products included not only phenyl propane (C₆—C₃) units but also C₆—C₂ and C₆—C₁ units. In addition, the infrared spectra suggested that the methanol-soluble portion maintains the typical structure of lignin, although it is rich in condensed-type linkages with some changes in the propyl side chain. These results indicate that the supercritical water treatment cleaves not only ether linkages but also part of the propyl chains in lignin to give various aromatic compounds.     

Quantifying lignin and holocellulose content in coniferous decayed wood using near-infrared reflectance spectroscopy

To determine the independent decomposition rates of lignin and cellulose of decayed woody debris, a technique for the rapid analysis of lignin and cellulose is required. We applied a near-infrared spectroscopy (NIRS) technique to measure the lignin and holocellulose content in decayed wood. We succeeded in creating partial least-squares (PLS) models to estimate the lignin and holocellulose content in the decayed wood of five species using NIR spectra. Although the accuracy was acceptable for the estimation of a five-species mixed model (R ² = 0.970 for lignin and R ² = 0.962 for holocellulose), it was further improved when the model was applied to each species independently. This combination of NIRS and a PLS model is a valuable tool for the determination of the lignin and holocellulose content in decayed wood. The technique is time efficient (3 min per sample) and non-hazardous (no acid treatment is required).     

Lignin fragments rich in detached side-chain structures found in water-soluble LCC

The content of the glyceraldehyde-2-aryl ether-type structure in water-soluble lignin–carbohydrate complex (LCC) fractions, which were obtained from Japanese cedar and birch residual wood meal after milled wood lignin isolation, was determined by ozonation. Quite high amounts of the glyceraldehyde-2-aryl ether-type structure were found in water-soluble LCC fractions of both species; about 3–5 times higher than those of other fractions. This result, as well as the high content of the β-1 structure in these fractions shown in our previous reports, suggest that lignin in these fractions has the characteristics of endwise-type lignin, because the glyceraldehyde-2-aryl ether-type structure and β-1 structure are typical characteristics of this type of lignin fraction. These results are in good agreement with the generally accepted hypothesis that the glyceraldehyde-2-aryl ether-type structure and β-1 structure are generated at the same time by a radical coupling reaction. The results also indicated that these two structures are present in close proximity in the lignin.     

Two-step hydrolysis of Japanese beech as treated by semi-flow hot-compressed water

A two-step hydrolysis of Japanese beech (Fagus crenata) was conducted by semi-flow treatment with hot-compressed water. The first treatment stage was conducted at 230°C/10 MPa for 15 min and the second at 270°C/10 MPa for 15 min. Hemicellulose and lignin were found to be hydrolyzed in the first stage, while crystalline cellulose was hydrolyzed in the second stage. The treatment solubilized 95.6% of the Japanese beech wood flour into water with 4.4% remaining as water-insoluble residue, which was composed mainly of lignin. Hydrolysis products from the first stage were xylose and xylo-oligosaccharides, glucuronic acid and acetic acid from O-acetyl-4-O-methylglucuronoxylan, and hydrolyzed monomeric guaiacyl and syringyl units and their dimeric condensed-type units from lignin. Products from the second hydrolysis stage were glucose and cello-oligosaccharides from cellulose. The dehydrated products levoglucosan, 5-hydroxymethylfurfural (5-HMF), and furfural, as well as fragmented products glycolaldehyde, methylglyoxal, and erythrose, were recovered in the first stage from hemicellulose, and to a greater extent in the second stage from cellulose. Furthermore, organic acids such as glycolic, formic, acetic, and lactic acids were recovered in both stages. Based on these lines of evidence, decomposition pathways of O-acetyl-4-O-methylglucuronoxylan and cellulose are independently proposed.     

Structural conversion of the lignin subunit at the cinnamyl alcohol stage in Eucalyptus camaldulensis

The lignin biosynthetic pathway in Eucalyptus camaldulensis was investigated by feeding stems with deuterium-labeled precursor. Pentadeutero[,-D₂ OCD₃] coniferyl alcohol was synthesized and supplied to shoots of E. camaldulensis, and incorporation of the labeled precursor into lignin was traced by gas chromatography-mass spectrometry. In addition to the direct incorporation of labeled precursor into the guaiacyl unit, a pentadeuterium-labeled syringyl unit was detected. This finding indicates that the -deuterium atoms in the hydroxymethyl group of labeled coniferyl alcohol remain intact during modification of the aromatic ring. The relative level of trideuterium-labeled syringyl monomer (the result of conversion via the cinnamic acid pathway) was negligible, suggesting that the pathway at the monolignol stage is used for conversion of exogenously supplied precursor. Our results provide conclusive evidence of a novel alternative pathway for generation of lignin subunits at the monolignol stage even in plants that do not accumulate coniferin in lignifying tissues.     

Contribution of lignin to the reactivity of wood in chemical modifications II: influence of delignification on reaction with vaporous formaldehyde

Participation of lignin in the reaction between vapor-phase formaldehyde and wood was examined by using gradually delignified wood meal. A fi rst-order rate equation was successfully applied to the weight gain data. From the estimated reaction parameters such as rate constant, k, and ultimate weight gain, a, the reactivity toward formaldehyde was discussed among wood components, and compared with that for acetylation. k decreased monotonously with progress of the elimination of lignin, suggesting that the reaction rate of lignin is dominant over that of whole wood, and the decrease in the ratio of lignin retarded the reaction of wood as a whole. On the other hand, a increased with decreasing lignin content. This may be attributable to the enhanced reactivity of the remaining lignin due to some structural changes and to the increase in the number of reactive sites in polysaccharides as a result of their exposure accompanying the elimination of lignin. The dependencies of k and a on the lignin content were not similar to the case for acetylation, probably because of the difference in the reaction phase. In vapor-phase formaldehyde treatment, the remaining lignin reacts as it is, whereas in liquid-phase acetylation it would undergo rearrangement or swelling of the structure in the reaction solution.