Fractional and structural characterization of ball-milled and enzyme lignins from wheat straw

Ball-milled and enzyme lignins were extracted with 90 and 50% dioxane–water from 6 days ball-milled wheat straw, and subsequently cellulase-treated straw residues, respectively. The crude lignin preparations were purified using a two-step precipitation method instead of the traditional ether precipitation procedure, and fractionated into pure milled lignin (PML), pure enzyme lignin (PEL), hemicellulose-rich milled lignin (HRML), lignin-rich enzyme lignin (LREL), and solubilized lignin during enzyme treatment (SLET) fractions. The five lignin fraction were studied using spectroscopic and degradative tecyhniques. The PML and PEL fractions showed very low content of associated polysaccharides (2.36–2.86%). The PML is mainly composed of β-O-4 ether bonds in the lignin structural units. The less common β-5 and β-β carbon–carbon linkages are also present. The results obtained also indicated that the lignins in wheat straw cell walls appeared to be very closely associated to p-coumaric and ferulic acid, and glucuronic acid or 4-O-methylglucuronic acid. © 1998 John Wiley & Sons, Inc. J Appl Polym Sci 68: 1633–1641, 1998     

Fractionation of Lignocellulosic Materials for the Biorefinery: Separation and Characterization of Lignin from Calamagrostis angustifolia Kom

Dewaxed Calamagrostis angustifolia Kom was pretreated with hot water at 60 and 90°C for 3 h, and then the residue obtained was successively treated with 70% ethanol, and 70% ethanol containing 0.2%, 1.0%, 2.0%, 4.0%, and 8.0% NaOH at 80°C for 3 h. The dissolved components were subjected to further separation to get eight lignin fractions, which were characterized by gel permeation chromatography, Fourier transform infrared, and sugar analysis. All the lignin fractions had small weight-average molecular weights between 810 and 2580 g/mol. Two typical lignins, L₃ (prepared with 70% ethanol) and L₅ (prepared with 70% ethanol containing 1.0% NaOH), were further analyzed using ¹H, ¹³C NMR and HSQC spectroscopy. Signals from guaiacyl (G), syringyl (S), and p-hydroxyphenyl (H) units observed in aromatic/olefinic region of HSQC spectra indicated that the lignin from Calamagrostis angustifolia Kom could be classified as “GSH” lignin. In aliphatic-oxygenated region, β-O-4′ together with small amounts of β-5′, β-β′, and p-hydroxycinnamyl alcohol end group were the main interunit linkages observed. Aqueous ethanol, which could avoid the cleavage of ether bonds in lignin at neutral condition, was more effective than water on lignin extraction.     

Quantitative Phosphorus-31 NMR Analysis of Six Soluble Lignins

By using a set of lignin samples, which have been subjected to thorough analyses by the international wood chemistry community, the recently developed quantitative method of ³¹P NMR spectroscopy was comprehensively examined. The values of total phenolic hydroxyl groups and those of total hydroxyl groups were found to favourably compare with those obtained by other laboratories, applying independent methods of analysis. Furthermore, the application of quantitative ³¹P NMR spectroscopy offered additional detailed structural information for the examined lignins which was in accord with literature accounts for similar wood species and lignin preparations. More specifically, the steam explosion lignins from aspen and yellow poplar woods and that produced by ball milling/enzyme hydrolysis of cottonwood were found to contain relatively high amounts of β-O-4 structures. In contrast, the kraft, organosolv, and the acid hydrolysis processes were found to induce significant chain scission on the resulting lignins. Ball milled cottonwood lignin contained the highest frequency of β-O-4 bonds and the lowest amount of free phenolic hydroxyls. The erythro form of β-O-4 structures were invariably predominant in the lignins from aspen, yellow poplar and cottonwood, in accord with the conclusions of previous reports on hardwood lignins. Thus, the application of quantitative ³¹P NMR spectroscopy offered the detailed chemical composition of the examined lignins.     

Fractional Separation and Structural Characterization of Chlorophyll and Lignin from Perennial Ryegrass (L. perenne) and Cocksfoot Grass (D. glomerata)

Abstract

One chlorophyll rich fraction and two lignin preparations were separated from perennial grass and cocksfoot grass by sequential three‐stage treatments with 80% ethanol containing 0.2% NaOH, 2.5% H₂O₂?0.2% EDTA containing 1.5% NaOH, and 2.5% H₂O₂?0.2% TAED containing 1.0% NaOH at 75°C for 3 h, respectively, which released 4.6 and 3.6% chlorophyll rich fraction, 2.3 and 5.8%, and 0.9 and 1.0% lignin preparations, except for releasing 8.0 and 10.4%, 79.1 and 77.0%, and 12.9 and 12.5% of the original hemicelluloses, respectively. The lignin fractions obtained from the two different grasses had very similar molecular weights and structural composition. The NMR spectra of the lignin preparations revealed the presence of p‐hydroxyphenyl, guaiacyl, and syringyl structures, and the lignin in chlorophyll rich fraction contained more guaiacyl and syringyl units than p‐hydroxyphenyl unit, whereas the reverse trend was found in the two lignin preparations. The lignin preparations are distinguished with straw and wood lignins by relatively higher contents of p‐hydroxyphenyl unit and lower amounts of condensed units (β‐5 and 5‐5′) and resinol units (β‐β). This difference in distribution of structural units indicated some structural heterogeneity between grass and straw/or wood lignin.     

BEHAVIOR OF EUCALYPTUS GLOBULUS LIGNIN DURING KRAFT PULPING. II. ANALYSIS BY NMR,ESI/MS,AND GPC

ABSTRACT

Residual and dissolved lignin from different phases of kraft delignification of Eucalyptus globulus wood were isolated and characterized by 1D and 2D ¹H NMR, ¹³C NMR, Electrospray Ionization Mass Spectrometry (ESI/MS), and gel permeation chromatography (GPC). During the temperature rise period, below 70°C, about 20% of the lignin was dissolved without significant structural changes. Above 70°C, the lignin suffered significant degradation/fragmentation in the cell wall prior to dissolution. The lignin ether-linked syringyl units were the most susceptible to alkaline degradation. Through the course of pulping, the residual lignin (RL) revealed a gradual increase of aliphatic moieties of unknown structure, as well as a decrease of native structures such as phenylcoumaran and pino-/syringaresinol lignin units. A significant decrease of the β-O-4 structures content in RL was detected only at the final cooking temperature. The lignin dissolved in the black liquor (BL) consisted of highly branched oligomers with rather low molecular weight (average mass 800–1000 u). A part of BL (about 30%) was chemically linked to carbohydrates and possessed a large molecular weight distribution (500–4000 u). BL showed a progressive decrease in β-O-4 and pino-/syringaresinol structures and formation of enol ether and stilbene structures. The GPC analyses showed a continuous decrease of the molecular weight of both the residual and dissolved lignins during the pulping process, particularly in the residual stage.     

Graft copolymers of lignin with electron poor alkenes

The synthesis of copolymers between lignin and electron poor alkenes is described. Lignin from steam‐exploded pine, from steam‐exploded straw, and organosolv were used as starting materials. Beforehand, lignins were fully characterized by using elemental analysis, ultraviolet spectroscopy, gel permeation chromatography (GPC), Fourier transform infrared (FTIR), and both¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy. The synthesis of copolymers was performed using a previously described procedure utilizing calcium chloride and hydrogen peroxide as reagents. FTIR of copolymers showed absorptions due to the presence of both lignin and the electron withdrawing group on the alkene. GPC analysis showed the presence of fractions with high molecular weights: the M_z of lignin from pine was 3729 while the copolymer with methyl acrylate showed M_z = 383790. Differential calorimetry showed the presence of glass transitions in the range of ?9 to 4.5°C due to the presence of grafted polyalkene chains. When acrylonitrile was used as starting material DSC analysis of the product showed a glass transition at 119°C, which can be attributed to grafted polyacrylonitrile chain. Lignin from steam explosion could be a good raw material in the preparation of graft copolymers. Furthermore, lignin from pine gave better results than that from straw. Finally, lignin from steam explosion gave better results than organosolv lignin. These results can be explained on the basis of the structural properties of used lignins. Both UV and ¹³C NMR spectra showed that lignin from pine contained a consistent amount of double bonds. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 1163–1171, 2003     

Extraction and characterization of lignins from maize stem and sugarcane bagasse

Lignins were isolated from maize stem and sugarcane bagasse by using mild dioxane or acidic dioxane solution. The result of nitrobenzene oxidation of the isolated lignins shows that there is a high proportion of p‐hydroxyphenyl alcohol in the lignins of maize stem and sugarcane bagasse. The lignins isolated from maize stem and sugarcane bagasse have relatively same value of the weight‐average (M _w = 3405–3868 g mol⁻¹) and number‐average (M _n = 1411–1612 g mol⁻¹) molecular weights, and polydispersity (M _w/M _n = 2.24–2.51). Acidic dioxane treatment did attack the β‐aryl ether structures in lignins, in particular for β‐aryl syringyl ethers, and broke the ester bonds between arabinose and ferulic acid that etherified to lignins, and it also cleaved lots of bonds in hemicellulosic polymer. The proportion of β‐O‐4 (threo) guaiacyl units is higher than that of β‐O‐4 (erthreo) guaiacyl units. The phenyl glycoside and benzyl ether linkages between lignin and hemicelluloses are also demonstrated in NMR analysis. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Gordon Research Conference on Chemicals and Materials From Renewable Resources

Abstract

This Conference will be held July 2-6, 1984 at Brewster Academy, Wolfeboro, New Hampshire. Among the topics to be discussed are the following: solid state ^iJC NMR spectra of celluloses, ultrastructure ofcellulose fibers - some new aspects, plant cell wall polysaccharides, 13 structural C NMR studies of lignin and lignin-like materials from flash utohydrolyzed wood, isolation of lignins from wood, electron transfer reactions in lignin and pulping chemistry, and structural inhomogeneity of lignin in wood cell walls.     

Fractional Isolation and Structural Characterization of Lignins from Oil Palm Trunk and Empty Fruit Bunch Fibers

Abstract

The lignins from dewaxed oil palm trunk and empty fruit bunch (EFB) fibers were fractionated into 95% ethanol soluble, cold and hot water soluble, and 1% NaOH soluble lignins, respectively. The chemical and structural composition of the lignin preparations was determined by using UV, GPC, FT-IR, ¹³C-NMR spectroscopy and nitrobenzene oxidation. The alkali soluble and 95% ethanol soluble lignin fractions were found to contain low amounts of chemically linked polysaccharides, 2.9-3.9% and 7.5-8.0%, respectively, while the water soluble lignin fractions showed significant amounts of bound polysaccharides (16.2-23.3%). All of the lignin fractions contained a high proportion of non-condensed syringyl units, together with small amounts of non-condensed guaiacyl and fewer p-hydroxyphenyl units. The lignin from oil palm EFB fiber contained a significant amount of esterified p-hydroxybenzoic acid and a minor quantity of esterified glucuronic acid. Trace of ferulic acids was both esterified and etherified to lignin side chains in the EFB fiber cell walls.     

BEHAVIOR OF EUCALYPTUS GLOBULUS LIGNIN DURING KRAFT PULPING. I. ANALYSIS BY CHEMICAL DEGRADATION METHODS

ABSTRACT

Eucalyptus globulus wood was subjected to kraft pulping, reaching different extents of delignification. The residual and dissolved lignins were isolated by soft acidolysis and acidic precipitation, respectively, and submitted to analysis of residual sugars and methoxyl groups, as well as to analysis by nitrobenzene and permanganate oxidation and thioacidolysis. Results from both residual and dissolved lignins analyses indicated that in the initial phase of pulping there is a preferential removal of lignin enriched in guaiacylpropane (G) and p-hydroxyphenylpropane (H) units, which is highly condensed and bonded with polysaccharides, predominantly with xylan. During the bulk and residual phases of pulping, lignin enriched in syringylpropane (S) units is progressively removed, leading to the increase of S/G ratio of black liquor lignin. After a purification step of dissolved lignins, two fractions with distinct levels of carbohydrates and with different structural features were obtained. The overall results were interpreted in terms of the topochemistry of the kraft pulping process.     

Fractional Separation of Hemicelluloses and Lignin in High Yield and Purity from Mild Ball-Milled Periploca sepium

Abstract

A novel three-step procedure for separation of hemicelluloses and lignin with high yield and purity was proposed in this study, where wood is mildly milled and successively extracted to produce three hemicellulosic and lignin fractions representing the total hemicelluloses and lignin in wood. The sequential treatments of the mild ball-milled Periploca sepium with 80% aqueous dioxane containing 0.05 M HCl at 85°C for 4 h, DMSO at 85°C for 4 h, and 8% NaOH at 50°C for 3 h resulted in a total release of over 85% of the original hemicelluloses and 86% of the original lignin. In particular, approximately 36% of the original hemicelluloses and 50% of the original lignin were separated during the first mild acidolytic hydrolysis process after low intensity milling. The structure of the acidic dioxane-, DMSO-, and alkali-soluble hemicellulosic and lignin fractions were elucidated using wet chemical analysis, FT-IR, and solution-state ¹H, ¹³C, and ³¹P NMR techniques. Results showed that both the mild ball milling and the mild acidolysis under the conditions given did not affect the separated lignin macromolecular structure. On the other hand, the mild acidolytic hydrolysis condition did cause substantial hemicellulosic depolymerization exception for a significant cleavage the ether linkages between lignin and hemicelluloses. The acidic dioxane-soluble lignin fraction was structurally different from the DMSO- and alkali-soluble lignin preparations and may originate mainly from the primary wall, while the alkali-soluble lignin preparation was mainly released from the secondary wall of Periploca sepium. Furthermore, it was found that the acidic dioxane-soluble hemicelluloses mainly contained more branched and less acidic arabinoxylans, and the 8% NaOH-soluble hemicellulosic fraction H₃ was both less branched and less acidic in structure, whereas the DMSO-soluble hemicelluloses were more acidic but less branched and consisted mainly of 4-O-methylglucuronoarabinoxylan.     

Effects of pretreatment temperature and alkali concentration on the composition of alkali-soluble lignins from wheat straw

A new rapid method of two step precipitation for isolation of alkali-soluble lignins with relatively free of polysaccharides from wheat straw was proposed in this study. The isolated alkali-soluble lignin fractions LA contained rather low amounts of neutral sugars (0.7–1.0%) and had low average molecular weights (960–1440 Da). The isolated lignin fractions LA contained roughly equal amounts of guaiacyl (G) and syringly (S) units with relatively fewer p-hydroxyphenyl (H) units, and appeared to be closely associated to phenolic acids and glucuronic acid or 4-O-methylglucuronic acid. The chemical compositions in each of the fractions are reported. The physico-chemical properties and structure features of these lignin fractions LA were characterized by UV-, IR-, ¹³C-NMR-spectroscopy, and alkaline nitrobenzene oxidation. © 1996 John Wiley & Sons, Inc.     

Lignin–phenol–formaldehyde resin adhesives prepared with biorefinery technical lignins

In this study, four biorefinery technical lignins were used to synthesize lignin–phenol–formaldehyde (LPF) resin adhesives with a proposed formulation that was designed based on accurate analysis of the active sites in lignin with ³¹P nuclear magnetic resonance (NMR). The properties of the LPF resin adhesives and the plywoods prepared with them were tested. The structural features and curing behavior of the LPF resin adhesives were thoroughly investigated by solution‐ and solid‐state ¹³C NMR. Results indicated that the proposed formulation exhibited favorable adaptability for all four of these technical lignins for synthesis of LPF resin adhesives. High‐performance plywood with low emissions of formaldehyde could be successfully prepared with the synthesized LPF resin adhesives. All the LPF resin adhesives exhibited similar structure and curing behavior with the commercial phenol–formaldehyde (CPF) resin adhesive. However, the LPF resin adhesives showed relatively higher curing temperatures as compared with the CPF resin adhesive. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42493.     

Structural characteristics of different softwood lignins according to 1D and 2D NMR spectroscopy

Abstract

Along with cellulose, lignin is one of the most common polymers of plant origin. Numerous studies show that the complex and irregular structure of lignin is still unknown. First of all, the existing problems are connected with the peculiarities of lignins as a natural chemical compound. In this paper the general regularities of the chemical structure of dioxane lignins isolated from the softwood – larch (Lárix sibírica), pine (Pinus cembra) and juniper (Juníperus commúnis). The studies were carried out using a combination of ¹³C, ³¹P and 2D NMR spectroscopy. According to HSQC spectra it was established that the main structural fragments are the structures of β-aryl ether, followed by phenylcoumarans, with lower amounts of other substructures. Signals belonging to a methyl substituted phenylcoumaron were first identified on the NMR spectra of softwood dioxane lignins. Signals related to the structure of 3′-O-methylated dihydroquercetin were found in the composition of larch dioxane lignin.     

木质素分级对其应用性能的影响

木质素是一种结构复杂的天然芳香类聚合物,功能基团丰富,具有良好的开发应用前景。工业碱木质素是当前木质素转化利用的主体,主要源于制浆黑液。由于原料来源、制浆工艺的影响,木质素多分散性高、结构与性能不一,致使相应的木质素基产品的均一性与稳定性较差。通过木质素分级可以获得具有不同分子量或特定化学结构特征的木质素级分,促使各级分的多分散性降低、均一性提高,进而针对特定性能的级分进行改性或者直接利用可进一步提高产品的稳定性。对现行木质素的主要分级方式进行了总结,重点探讨了工业碱木质素的分级方式对其后续利用的影响及存在问题。最后,针对木质素不同级分的特点对木质素未来分级的方向及应用前景提出了展望。     

Structural characterization of residual lignins isolated with cyanamide-activated hydrogen peroxide from various organosolvs pretreated wheat straw

The posttreatment of various organosolvs pretreated wheat straw with cyanamide-activated hydrogen peroxide was studied. About 44–80% of the total residual lignin and 38–85% of the total residual hemicelluloses were released or degraded during the posttreatment with 1.8% H₂O₂–0.18% cyanamide at 50°C under pH 10.0 for 4 h from different aqueous organic acids or alcohols pretreated straw. The seven degraded residual lignin preparations were subjected to a comprehensive physicochemical and structural characterization by UV, FTIR, and ¹H and ¹³NMR spectroscopy, and GPC. The nitrobenzene oxidation method was also applied to the in situ lignins. It was found that the seven residual lignin preparations contained large amounts of noncondensed syringyl and guaiacyl units, together with fewer noncondensed p-hydroxyphenyl units, esterified p-coumaric acid, and mainly etherified ferulic acid. All of the lignin fractions are free of associated polysaccharides and had molecular-average weights ranging between 2980 and 3820 g mol⁻¹. Analysis of these low molecular weight degradation products revealed an oxidation of residual lignin had occurred. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Lignin and Other Aromatic Substances Released from Spruce Wood During Pressurized Hot-Water Extraction,Part 2: Structural Characterization

Extraction of ground spruce sapwood with pressurized hot water in an accelerated solvent extractor (ASE) at 170°C during 20, 60, and 100 min resulted in isolation of galactoglucomannans and aromatic substances, including lignin. The isolated lignin preparations were characterized by spectrometric (UV, FT-IR, ¹H NMR, liquid and solid-state ¹³C NMR), chromatographic (RP-HPLC, HP-SEC, GC-FID, and GC-MS), conventional pyrolysis, thermally assisted hydrolysis, and methylation techniques in tandem with GC-MS, and classical wet chemistry (methoxyl groups, total and phenolic hydroxyl groups, derivatization followed by reductive cleavage—DFRC). The content of β-O-4 bonds in isolated lignins was similar to that in MWL and their proportion decreased with extraction time. The oxidation of isolated lignins and content of total hydroxyl groups were significantly increased with extraction time. The lignin structure underwent condensation and demethylation reactions during hot-water extraction. The induction of new phenylcoumaran substructures was proposed in isolated lignins.     

How Well Do MWL and CEL Preparations Represent the Whole Hardwood Lignin?

Hardwood lignin preparations were isolated using classical milled wood lignin (MWL) and cellulolytic enzyme lignin (CEL) protocols. Furthermore, we managed to produce a lignin preparation of a very high yield, above 90%, with high purity and minimal structural degradation. This was achieved by dissolution and regeneration of milled wood prior to enzymatic hydrolysis, along with the use of 80% dioxane for lignin extraction. This preparation (RCEL-80) yield was about 4.5 and 2.5 times higher than the yields of the traditional MWL and CEL preparations, correspondingly, at the same milling time. The preparations obtained were comprehensively analyzed with state-of-the-art quantitative NMR techniques and wet chemistry methods. CELs were representative preparations for hardwood lignins as the S/G ratios in the CELs were equivalent to those in lignin in situ. Degradation of the main lignin subunits was very low. Importantly, the structures of CELs were independent of the preparation yield and were very similar to the structure of the high-yield lignin, RCEL-80. In contrast, the structures of MWLs were noticeably dependent on the preparation yield, especially when the yield was below 15–20% of total wood lignin. In particular, the S/G ratio increased with increasing MWL yield, but was still lower than the S/G ratios of the whole lignins in situ, even at high MWL yields. The amounts of β-O-4 lignin units in MWL were lower than those in the corresponding CEL preparations. It has been concluded that CEL preparations were representative of the whole wood lignin whereas MWLs represent a fraction with a lower S/G ratio.     

Investigation of oil palm based Kraft and auto-catalyzed organosolv lignin susceptibility as a green wood adhesives

The aim of this study is to highlight the application and potentiality of oil palm based lignins in the synthesis of green phenolic resins. The delignification processes were conducted using Kraft and auto-catalyzed ethanol–water pulping processes. The extracted lignins were characterized using elemental analysis, Fourier transform-infrared, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy, molecular weight distribution (M_n, M_w and polydispersity), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). The obtained FTIR results revealed that the Kraft lignin contained substantial amounts of guaiacyl units with smaller amounts of syringyl units. The molecular weight of Kraft lignin was 1564 g mol⁻¹ which is higher than organosolv lignin at 1231 g mol⁻¹. The activated free ring position (2.99%) of Kraft lignin was comparatively higher than that of organosolv lignin (2.06%) which was measured using Mannich reactivity analysis. Thermal analysis of Kraft lignin showed higher thermal stability compared to organosolv lignin. The structural and thermal characteristics implied that Kraft lignin had higher potential for the production of green phenolic resins when compared with organosolv lignin.     

Structure and Reactivity of Spruce Mechanical Pulp Lignins Part II. Organosolv Fractionation of Lignins in a Flow-Through Reactor

Various spruce mechanical pulps were subjected to delignification with ethanol-water (1 : 1, v / v ) containing 0.1 M acetic acid at 175°C in a flow- through reactor. A thermomechanical pulp (TMP) and the corresponding samples derived from its bleaching (BTMP) and yellowing ( YBTMP ) treatments were delignified to a similar extent, about 70% of delignification degree, as compared with spruce wood. A series of five successive lignin fractions was recovered from each pulp sample and then characterized by various analytical methods. Large structural variations were observed within these fractions. The number of phenolic structures appeared to be an important factor influencing the dissolution of lignins in the ethanol-water medium. These lignin fractions were found to be different in the amount of β-aryl ether structures and in the relative importance of main carbon-carbon bonding patterns. The results are discussed in relation to the lignin fractionation in the flow-through reactor.