Comprehensive Evaluation and Prediction of the Effectiveness of H2O2-assisted Na2CO3 Pretreatment of Corn Stover Using Multivariate Analysis

In this study, multivariate analysis methods, including a principal component analysis (PCA) and partial least square (PLS) analysis, were applied to reveal the inner relationship of the key variables in the process of H₂O₂-assisted Na₂CO₃ (HSC) pretreatment of corn stover. A total of 120 pretreatment experiments were implemented at the lab scale under different conditions by varying the particle size of the corn stover and process variables. The results showed that the Na₂CO₃ dosage and pretreatment temperature had a strong influence on lignin removal, whereas pulp refining instrument (PFI) refining and Na₂CO₃ dosage played positive roles in the final total sugar yield. Furthermore, it was found that pretreatment conditions had a more significant impact on the amelioration of pretreatment effectiveness compared with the properties of raw corn stover. In addition, a prediction of the effectiveness of the corn stover HSC pretreatment based on a PLS analysis was conducted for the first time, and the test results of the predictability based on additional pretreatment experiments proved that the developed PLS model achieved a good predictive performance (particularly for the final total sugar yield), indicating that the developed PLS model can be used to predict the effectiveness of HSC pretreatment. Therefore, multivariate analysis can be potentially used to monitor and control the pretreatment process in future large-scale biorefinery applications.     

Effect of chemical pretreatments on the fermentation and ultimate digestibility of bagasse by Phanerochaete chrysosporium

When sugarcane bagasse was pretreated at room temperature with various chemicals singly or in combination, (0.25 m NaOH; 0.5 m H₂SO₄; 2.6 M NH₃;0.14 M Ca(OH)₂+0.12 M Na₂CO₃;4.2 m EDA; and 0.5 M H₂SO₄ or 0.25 M NaOH followed by 4.2 M EDA), increases in in-vitro digestibility range from 28.9 to 48.6% depending on the particular chemicals. With most pretreatments, the hemicellulose content decreased and the cellulose content increased slightly, while lignin levels decreased only with NaOH + EDA pretreatments. Further fermentation of the chemically pretreated bagasse with Phanerochaete chrysosporium gave increased digestibilities, the highest final digestibility (59%) being achieved by pre-treatment with 0.25 M NaOH. Pretreatment with NaOH or Ca(OH)₂+Na₂CO₃ and fermentation gave greatly increased levels of hemi-cellulose and cellulose and substantial decreases in lignin levels. Chemical pretreatments of bagasse together with autoclaving considerably enhanced in-vitro digestibility (up to 75%) and lignin degradation (1.4%). Pretreatment with NaOH, Ca(OH)₂+Na₂CO₃, and EDA plus autoclaving gave large increases in total hemicelluloses and cellulose and marked decreases in lignin. Further fermentation with P. chrysosporium gave only slight increases in digestibility.     

杨木APMP制浆化学预处理废液中有机物的分析

研究了不同化学预处理条件对APMP废液中木素和糖类物质浓度的影响。结果表明,化学预处理过程中化学药剂NaOH用量、H₂O₂用量和Na₂SiO₃用量对化学预处理废液中木素和糖类物质浓度的变化有不同影响,影响程度由大到小依次为NaOH用量＞H₂O₂用量＞Na₂SiO₃用量,其中NaOH用量和H₂O2用量对木素和糖类物质浓度的变化有重要影响,Na₂SiO₃用量无明显影响。化学预处理温度对化学预处理废液中木素和糖类物质浓度的影响程度明显大于化学预处理时间。在化学预处理过程中,NaOH用量、H₂O₂用量和化学预处理温度是影响杨木APMP制浆化学预处理废液中木素和糖类物质浓度的重要因素。     

Orientierende Untersuchungen zum Organosolv-Aufschluß von Buchenholz mit einem Propandiol/Wasser-Gemisch

Beech wood (Fagus sylvatica L.) was digested in organosolv processes using ethanol/water (EtOH/H₂O) and propandiol-(1,2)/water (PDO/H₂O). The pulping qualities of both agents were compred. The pulp yield is lower for PDO/H₂O than for EtOH/H₂O. While almost the whole lignin can be recovered in a simple manner from the EtOH spent cooking liquor, the lignin from the PDO-liquors can only be partly isolated. Due to its high boiling point the recovery of PDO from the spent pulping liquors is more difficult than that of EtOH. Schemes of the recovery of pulps and lignins as well as lignin and carbohydrate balances demonstrate the results of the pulping experiments.     

Technical note: A comparison of alkali treatment methods to improve neutral detergent fiber digestibility of corn stover

The objective of this study was to compare neutral detergent fiber (NDF) digestibility of corn stover that had been treated by 2 alkali treatment methods. Two experiments were conducted to test a sodium hydroxide (NaOH) treatment method that uses an ethanol/water co-solvent (NaOH/ethanol-H₂O, United States Patent No. 20140220228) and a calcium hydroxide (CaOH) treatment method, which uses water as a solvent (CaOH/H₂O). An in situ trial was conducted to compare NDF digestion kinetics between NaOH/ethanol-H₂O-treated stover, CaOH/H₂O-treated stover, untreated corn stover, and soy hulls. The digestion rate of potentially digestible NDF (k_d) of NaOH/ethanol-H₂O-treated corn stover (5.36%/h) was higher than CaOH/H₂O-treated stover (2.27%/h), or untreated corn stover (1.76%/h) and similar to the k_d of soy hulls (4.93%/h). The indigestible NDF (iNDF) fraction of untreated corn stover (35.1% of NDF) was reduced by CaOH/H₂O treatment (27.3% of NDF) and by NaOH/ethanol-H₂O treatment (2.8% of NDF). The iNDF fraction in soy hulls (3.6% of NDF) was similar to iNDF of NaOH/ethanol-H₂O-treated stover. An in vivo digestibility trial was also conducted to compare fiber digestibility of diets supplemented with untreated corn stover, NaOH/ethanol-H₂O-treated corn stover, or soy hulls. Total-tract apparent dry matter (DM) and NDF digestibility were measured with 8 lactating Holstein cows in a replicated 4 × 4 Latin square with four 21-d periods. Apparent DM digestibility (DMD) was improved when supplemental soy hulls were added to the base diet (60.0% DMD) compared with the base diet with no supplemental fiber (57.7% DMD). Apparent DM digestibility was reduced when diets were supplemented with untreated stover (52.4%). Dry matter digestibility of NaOH/ethanol-H₂O-treated stover was similar (54.8% DMD) to all other treatments. Digestibility of NDF was lowest when cows were fed the diet with supplemented untreated stover (35.5% of NDF), and improved when soy hulls (40.6% of NDF) or NaOH/ethanol-H₂O-treated stover (43.8% of NDF) were added to the diets. The NaOH/ethanol-H₂O treatment process improves the DM and NDF digestibility of corn stover to values similar to those of soy hulls.     

Sodium Chlorite Treatment of Plant Materials: Fiber and Lignin Composition,Digestibility, and Their Interrelationships

Effects of NaClO₂ treatment on various structural components of different feedstuffs were studied. Six hays (alfalfa, tall fescue, orchardgrass, red clover, timothy, and a mixed hay), two straws (barley and wheat), four stovers (two corn and two soybean), three hulls (peanut, rice, and soybean), and corn cobs were treated with sodium chlorite at 11 concentrations ranging from .000 to .394 g of sodium chlorite/g of feed. At .394 g, NDF fiber and organic dry matter percent digestibilities were alfalfa hay, 93, 98; barley straw, 96, 98; corn cobs, 98, 99; corn stover I, 100, 100; corn stover II, 98, 99; tall fescue hay, 95, 97; mixed hay, 93, 96; orchardgrass hay, 93, 96; peanut hulls, 15, 42; red clover hay, 88, 95; rice hulls, 64, 79; soybean hulls, 98, 99; soybean stover I, 97, 98; soybean stover II, 97, 98; timothy hay, 95, 96; and wheat straw, 99, 99. Although NDF content (organic matter basis) was significantly correlated with concentration of NaClO₂ for all feeds, ADF content (organic matter basis) was not for soybean hulls and lignin content was not for peanut hulls. Regression analysis of lignin oxidation products against concentration of NaClO₂ showed a significant correlation for syringaldehyde when using all feeds. Individually, only for corn cobs was syringaldehyde highly correlated with treatment. Regressions between lignin oxidation products and digestibility measurements likewise varied with the feed and with the specific measure of digestibility.     

Influence of Chemical Pretreatment on the Dissolved Organics in Poplar Alkaline Peroxide Mechanical Pulping Effluent

The components of the effluent from the chemical pretreatment of poplar alkaline peroxide mechanical pulp (APMP) were analyzed in this study. The main dissolved organics were low-molecular weight (LMW) lignin, oligosaccharides, and monosaccharides. The lignin and sugar concentrations in the effluent obtained using different chemical pretreatment conditions and chemical dosages were analyzed using ultraviolet (UV) spectroscopy. This analysis provided a theoretical basis for the subsequent treatment and utilization of APMP effluent. The experimental results showed that the dosages of NaOH, H₂O₂, and Na₂SiO₃ in the chemical pretreatment process affected the lignin and sugar concentrations in the effluent and that different chemicals had differing degrees of influence. The degree of influence exhibited the following order: NaOH>H₂O₂>Na₂SiO₃. More specifically, the dosages of NaOH and H₂O₂ had stronger influences on the lignin and sugar concentrations than that of Na₂SiO₃. Indeed, the Na₂SiO₃ dosage hardly affected the lignin and sugar concentrations in the effluent, but Na₂SiO₃ could stabilize the chemical pretreatment system and improve the reactive efficiency of NaOH and H₂O₂. The pretreatment temperature and time also affected the organic components, and the influence of the temperature was stronger than that of time.     

The effect of feeding pods of multipurpose trees (MPTs) on the degradability of dry matter and cell wall constituents of maize stover and alfalfa incubated in the rumen of sheep

This study evaluated the effects of feeding alfalfa and pods of tree legumes on rumen pH and ammonia concentration as well as in sacco degradability of dry matter and fibre constituents of maize stover, alfalfa hay and their neutral detergent extracts. The feeds were incubated in the rumens of five South African Merino sheep fed individually in an incomplete (5 × 4) Latin square design using five diets. The diets comprised equal proportions of pasture hay and either alfalfa (Alfalfa diet) or pods of Acacia sieberiana (Sieberiana diet), Acacia nilotica (Nilotica diet) or Leucaena leucocephala (Leucaena diet). The fifth diet was composed of pasture hay alone (Hay diet) and served as a negative control. Supplementation of hay with either legume pods or alfalfa significantly (p < 0.05) increased rumen ammonia concentration from 56 to a maximum of 86 mg l^?1. The pH of the rumen fluid ranged between 6.2 and 6.5 for all diets, but variations in pH were not significant. Diet did not affect (p > 0.05) the degradability constants lag time (LT), slowly degradable fraction (B), potential degradability (PD), effective degradability (ED) and rate of degradation (C) of dry matter (DM) but significantly (p < 0.01) affected ED of the fibre constituents of the incubated feeds. The effective degradabilities of the incubated feeds differed (p < 0.001), with alfalfa and maize stover having average values of 555 and 318 g kg^?1 DM respectively. The rate of degradation of alfalfa was also higher (p < 0.01) than that of maize stover. Neutral detergent extraction improved ED of the cell wall constituents of maize stover but produced the opposite effect for alfalfa. The interactions between incubation feed and extract were significant (p < 0.01) for ED and C of neutral detergent fibre and hemicellulose. The observations show that pods from tree legumes are comparable to alfalfa in the provision of rumen ammonia but limit microbial activity in the degradability of fibre constituents. They also show that maize stover and alfalfa differ in their degradabilities and that the differences may be attributed to their cell wall chemistry. © 2001 Society of Chemical Industry     

Evaluation of the prebiotic potential of five kiwifruit cultivars after simulated gastrointestinal digestion and fermentation with human faecal bacteria

We studied the in vitro prebiotic potential of five different cultivars of kiwifruit including the green‐fleshed ‘Hayward’ and ‘Zesh004’ and the gold‐fleshed ‘Hort16A’, ‘Zesy002’ and ‘Zesy003’. The kiwifruit (25 g fresh weight equivalent) were subjected to simulated gastrointestinal digestion before fermentation for 16 h with faecal microbiota from ten individual donors. Microbial metabolites including lactate were quantified while changes in microbiome composition were determined by 16S rRNA sequencing. Lactate concentrations were highest with ‘Hayward’ (P = 0.01) and correlated with the amount of the kiwifruit fibre and polyphenols, chlorogenic acid and cryptochlorogenic acid. All the kiwifruit behaved similar to inulin in increasing the relative abundance of Bifidobacterium (P < 0.001), but unlike inulin, significantly (P < 0.001) increased the abundances of Ruminococcaceae and decreased Bacteroides. In comparison with inulin, the green‐fleshed kiwifruit selectively increased Lachnospira (P = 0.008) while the gold‐fleshed kiwi fruit increased Akkermansia (P < 0.001). These data suggest that the fibre and polyphenol content of the kiwifruit play a role in modulating gut microbial metabolism. Further clinical studies with these kiwifruit cultivars are required to confirm the potential prebiotic benefits that may be achieved by normal dietary intervention.     

Chemical,nutritive, deoxynivalenol and zearalenone content of corn relative to the site of inoculation with different isolates of Fusarium graminearum

Kernels from two samples of 18 ears of corn were hand-shelled into three fractions (Fl, F2 and F3) relative to the site (F3) where the ears had previously been inoculated with one of four isolates of Fusarium graminearum (Schwabe). F3 and Fl were from the nearest and remotest parts of the ear, respectively, to the inoculation site. Ears of uninoculated control corn were processed in the same manner. The resulting 30 samples were analysed, after grinding (20 mesh), for deoxynivalenol (DON), zearalenone (ZEA), ergosterol (EGS), dry matter (DM), 200 kernel weight, Kjeldahl nitrogen (N), ether extract, ash, sugar, starch, acid and neutral detergent fibre (ADF, NDF, respectively), cellulose, hemicellulose and lignin. The true metabolisable energy (TME), TME corrected to zero N balance (TME_n), true available amino acid (TAAA) and true DM digestibility (TDMD) contents of these samples and corn to which 4, 16 or 64 mg purified DON kg^?1 were added, were determined using adult White Leghorn chickens. The DON, ZEA, EGS, N and ash contents were higher (P<0.001), and ether extract, TME and TME_n values were lower (p<0.01) in F3 than in Fl. Higer avilabilites accured for aspartic and glutamic acids, isoleucine, leucine and histidine in F2 and F3 than F1 (P<0.05). Carbohydrate and fibre values varied little (p>0.05) among fractions either within or among incoulation treatments. Sources of Fusarium inoculate influenced (P<0.05) the DON, ZEA, N, fat and TDMD of the corn. The TME_n and total TAAA of corn decreased (P<0.001) when purified DON was added. Moderate relationships (r=0.57?0.73) occurred between DON and ZEA, EGS and N. In general, these results are similar to those for wheat, and indicate that chemical and nutritive content of feedingstuffs may be increased when they are infected by toxigenic fungi.     

Phenolics and in-vitro degradability of protein and fibre in West African Browse

Relationships among soluble phenolics, soluble and insoluble proanthocyanidins (PAC), lignin, N, neutral-detergent fibre (NDF), and in-vitro degradability of protein and NDF were determined in 72 West African fodder trees and shrubs. Species were collected in the semi-arid (Niger), sub-humid (Nigeria) and humid/sub-humid (Benin) zones. Variation among species in chemical composition and in-vitro degradability of protein and NDF was large. Zones did not differ in mean content of phenolic compounds. Protein degradability was negatively correlated with soluble phenolics (r = ?0.34, P < 0.01) and soluble PAC (r = ?0.47, P < 0.001). NDF was positively correlated with soluble PAC (r = 0.44, P < 0.001), insoluble PAC (r = 0.28, P < 0.05) and lignin (r = 0.76, P < 0.001). NDF degradability was negatively correlated with soluble PAC (r = ?0.40, P < 0.001) and lignin (r = ?0.59, P < 0.001). Chemical composition and in-vitro degradability along with field observations can provide useful criteria for determining the nutritive value of browse species.     

Dietary fibre level and enzyme inclusion affect nutrient digestibility and excreta characteristics in grower pigs

An experiment was conducted with 12 grower pigs to determine the effects of (1) manipulating dietary neutral detergent fibre (NDF) level and (2) dietary inclusion of fibre‐degrading enzymes on nutrient digestibility and excreta characteristics in pigs. Four diets were formulated to contain three levels of NDF: 1, 66 g kg^?1 NDF (low fibre); 2, 121 g kg^?1 NDF (moderate fibre); 3, 222 g kg^?1 NDF (high fibre); 4, diet 3 plus 2.0 g kg^?1 enzyme. Increasing levels of dietary NDF linearly decreased dry matter and energy digestibilities (r² = 0.99, P < 0.05). Lowering dietary NDF from 222 to 121 g kg^?1 improved nitrogen digestibility by 13% (P < 0.01). Faecal production was decreased by 9% for each 1% decrease in dietary NDF content (r² = 0.99, P < 0.05). Inclusion of fibre‐degrading enzymes in the high‐fibre diet improved dry matter and energy digestibilities by 2 and 3% respectively, and reduced faecal production by 10% (P < 0.01). Faecal and manure (faeces plus urine) pH values from pigs fed the high‐fibre diet and the high‐fibre plus enzyme diet were lower than those from pigs fed the other experimental diets (P < 0.05). In conclusion, lowering the dietary NDF level or inclusion of fibre‐degrading enzymes in high‐fibre diets may offer relatively practical and easy methods for reducing waste production in pigs. © 2002 Society of Chemical Industry     

The relationships between gas production and chemical composition of 23 browses of the genus Sesbania

This study evaluated the differences in the gas produced by Sesbania accessions and related gas production to macro-constituents and tannin content of the browse species. Fresh samples were incubated in triplicate (about 200 mg per syringe) in a medium prepared from sieved rumen liquor harvested from two rumen-fistuiated cows fed native pasture hay ad libitum and 2.0 kg cottonseed cake per head per day. Sesbania species had similar rates of gas production but differed (P < 0.001) in the volume of gas produced. The ranking order for gas production was S grandiflora > S sesban > S keniensis > S goetzei. There was little variation in the gas production parameters within S grandiflora and S goetzei. But the variation was large within S sesban. Regression analysis showed that the rate of gas production was negatively affected (P < 0.025) by the lignin content of the browses, but the volume of gas produced was negatively related to the neutral detergent fibre, hemicellulose, lignin and insoluble proanthocyanidins content of browses. Ouster analysis showed a broad agreement between gas production constants, macro-constituents and tannin contents in classifying accessions into high or low nutritive value groups.     

Active packaging of ground beef patties by edible zein films incorporated with partially purified lysozyme and Na2EDTA

In this study, antimicrobial activity of zein films incorporated with partially purified lysozyme and disodium ethylenediaminetetraacetic acid (Na₂EDTA) has been tested on selected pathogenic bacteria and refrigerated ground beef patties. The developed films containing 700 μg cm^?2 lysozyme and 300 μg cm^?2 Na₂EDTA showed antimicrobial activity on Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhimurium. The application of lysozyme and Na₂EDTA incorporated zein films on beef patties significantly decreased total viable counts (TVC) and total coliform counts after 5 days of storage compared to those of control patties (P < 0.05). Zein films incorporated with lysozyme and Na₂EDTA or Na₂EDTA alone significantly slowed down the oxidative changes in patties during storage (P < 0.05). Redness indices of patties coated with zein films were significantly lower than those of uncoated control patties during storage (P < 0.05). This study demonstrated the potential usage of zein films containing lysozyme and Na₂EDTA for active packaging of refrigerated meat products.     

Salmonella typhimurium Attached to Chicken Skin Reduced Using Electrical Stimulation and Inorganic Salts

Chicken skins inoculated with Salmonella typhimurium at 1 × 10⁸ CFU/mL were subjected to chemical or electrical treatments (4 mA/ cm² current, 1 kHz frequency and 50% duty cycle) for 10 min in 1% sodium chloride (NaCl), sodium carbonate (Na₂CO₃), or trisodium phosphate (Na₃PO₄ 12H₂O or TSP). Salmonellae on chicken skins and in treatment solutions and rinsing water were enumerated with microbiological platings. Chicken skin was also examined using scanning electron microscopy.S. typhimurium attached to skins were reduced by 90% after electrical treatments in 1% NaCl, Na₂CO₃, or TSP, while the reduction ranged from 34% to 76% in groups treated by the salts alone.     

Lignification of switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii) plant parts during maturation and its effect on fibre degradability

Five cultivars of switchgrass (Panicum virgatum L) and four cultivars of big bluestem (Andropogon gerardii Vitman) were harvested at vegetative, boot and heading stages of maturity. Leaf and stem fractions were analysed for detergent fibre composition and 48-h ruminal in-vitro degradability, ester- and ether-linked non-core lignin phenolic acids, and core lignin composition. Big bluestem leaves contained more neutral detergent fibre than switchgrass, but general composition of the fibre did not differ. Stem fibre of switchgrass had relatively lower levels of cellulose and lignin at the vegetative stage than observed in big bluestem. Esterified and etherified p-coumaric and ferulic acid concentrations were generally higher in switchgrass plant parts. Yield of nitrobenzene oxidation products from core lignin was greater for switchgrass leaves, but very little difference in composition was noted. Leaf tissue contained lower concentrations of all lignin components than stems. Maturation resulted in increased total lignification, but all components did not respond in the same manner. Variation for all measures of lignification seemed to be as great within species as between the grass species. Degradability of fibre declined with maturation. The only species difference was that switchgrass fibre was more degradable at the vegetative stage. Relationships between lignification and fibre degradability were in agreement with some, but not all, previously reported relationships. Concentration of core lignin was only a significant predictor of fibre degradability when the relationship was examined across maturity stages. Within a stage of maturity, lignin composition was more closely related to fibre degradability than was concentration. However, the best predictors of fibre digestibility differed among species, plant part and maturity.     

漆酶对不同漂段纸浆催化氧化木素的效果研究

以未漂硫酸盐桉木浆(Sp浆)、氧脱木素桉木浆(O浆)、H_2O_2漂白桉木浆(P浆)作为不同漂段原料,探讨漆酶-谷氨酸体系对不同漂段纸浆的催化效果。结果表明,漆酶催化总体上可使纸浆的卡伯值增大,纸浆的白度随之下降;漆酶协同谷氨酸可催化氧化纤维表面木素与相邻纤维上的木素发生自由基偶合反应,同时使纤维表面的微细纤维暴露出来,两者共同作用使相邻纤维形成更好的结合力,使纸浆的强度性能大幅度提升;谷氨酸可通过漆酶催化氧化作用连接到木素上,增加纤维原料中的羧基含量,Sp浆、O浆、P浆羧基含量分别较未处理纸浆提高20%、50%、43%。     

Preparation and characterisation of dietary fibre from sugar beet pulp

Dried sugar beet pulp was ground and washed with 95% ethanol in order to obtain a colourless and odourless product which could be used as dietary fibre after drying and sieving. Sugar beet fibre had the same chemical composition as the initial pulp: 26–32% of hemicelluloses, 22–24% cellulose, 21.5–23% uronic acids, ～1–2% lignin, ～7–8% protein, 7.5–12% ash and ～0.5% residual sucrose. The main physicochemical properties such as density, cation exchange capacity (～0.6 meq g^?1) and water holding capacity (24–32 g g^?1) were determined. The water holding capacity of sugar beet fibre was only slightly affected by pH and ionic strength (adjusted by NaCl or CaCl₂) but it held less water in the H⁺ form than in the salt form, Na⁺ or Ca²⁺.     

Binding of Iron by Cellulose, Lignin, Sodium Phytate and Beta-Glucan, Alone and in Combination, Under Simulated Gastrointestinal pH Conditions

Cellulose (Ce), Lignin (L), Na phytate (P) and beta-glucan (G), alone, and in combination, were evaluated as to their effect on iron profiles in model systems containing FeSO₄• 7H₂O under simulated gastrointestinal pH conditions. Both cellulose and lignin alone, were found to bind a high percentage of the iron in an insoluble form at pH 5.7±0.4 and pH 6.6±0.1, but less at pH 2.0±0.1. However, in the presence of other model components, cellulose unlike lignin had a minimal effect on total binding at all pH levels. Na phytate, alone, formed soluble complexes at all pH levels and when added to lignin led to the solubiization of some of the iron bound to lignin. Beta-glucan formed soluble iron complexes, which appeared to have a low binding strength with iron.     

Lignin in the ‘cell contents’ fraction of tropical forages

In several tropical forage species the neutral detergent extraction used to determine cell wall and cell contents fractions solubilised a substantial UV‐absorbing polymeric fraction which appeared to be a soluble lignin–carbohydrate complex. A crude product was isolated from spear grass (Heteropogon contortus) with a yield of 30 g kg^?1. In other forages an acetyl bromide method was used to estimate lignin extractable by neutral detergent. For eight species of tropical grasses this gave the following values: Aristida calcyna, 38; Mitchell grass (Astrebla squarrosa), 37; Chloris gayana, 31; Eragrostis sp, 37; spear grass, 49; Iseilema membranaceum, 35; Panicum maximum, 36; Sorghum sp, 42; Themeda triandra, 79 g kg^?1. The levels in two legumes were: Alysicarpus bupleurifolius, 29; Macroptilium atropurpureum, 27 g kg^?1 dry matter. Sequential extractions on spear grass and Mitchell grass showed that most of the 50% dioxan‐soluble fraction was also soluble in neutral detergent, raising the possibility that ‘dioxan lignin’, on which studies of forage lignin structure in other species have been conducted, would be found in the cell contents fraction rather than in the fibre fractions on which lignin analyses have been performed. Addition of dioxan extractives reduced the in vitro neutral detergent fibre (NDF) digestibility of spear grass and Mitchell grass. © 2002 Society of Chemical Industry