Distribution and Chemical Composition of Lignin in Secondary Xylem of Cactaceae.

Cactaceae family has heterogeneity in the accumulation of lignocellulose due to the diversity of shapes and anatomy of the wood. Most studies focus on fibrous and dimorphic species; but the non-fibrous species are poorly studied. The aims of this work were to analyze the syringyl/guaiacyl ratio of lignin and its distribution in secondary xylem, especially in non-fibrous species. The syringyl/guaiacyl (S/G) ratio was quantified from 34 species of cacti by nitrobenzene oxidation of free-extractive wood. The distribution of lignocellulose in wood sections stained with safranin O/fast green was determined with epifluorescence microscopy. The S/G ratio was heterogeneous; most of the non-fibrous species had a higher percentage of syringyl, while the fibrous ones accumulate guaiacyl. Fluorescence emission showed that vessel elements and wide-band tracheids had similar tonalities. It is hypothesized that the presence of a higher percentage of syringyl in most cacti is part of the defense mechanism against pathogens, which together with the succulence of the stem represent adaptations that contribute to survival in their hostile environments.

Characterization of a lignin from Crataeva tapia leaves and potential applications in medicinal and cosmetic formulations.

Lignins are phenolic macromolecules that have several applications. In this work, we examine some biological activities of a lignin-like macromolecule isolated from the Crataeva tapia leaves, not yet studied to evaluate its potential applications in medicinal and cosmetic formulations. Lignin was obtained by alkaline delignification and its physical-chemical characterization was made by means of FT-IR, UV-Vis, NMR spectroscopy, elementary analysis, molecular mass determination and thermal analysis. Lignin is of the GSH type, with levels of hydrogen (5.10%), oxygen (27.18%), carbon (67.60%), nitrogen (0.12%) and phenolic content of 189.6 ± 9.6 mg GAE/g. In addition, it is a thermally stable macromolecule with low antioxidant activity. Cytotoxicity and cytokine production were assessed by flow cytometry. The photoprotective activity was evaluated by adding different concentrations of lignin to a commercial cream. Lignin was not cytotoxic, it stimulated the production of TNF-α, IL-6 and IL-10 and did not promote a significant change in nitric oxide levels. In addition, this macromolecule was able to promote increased absorption of ultraviolet light from a commercial cream. These results reinforce the ethnopharmacological use of C. tapia leaves and suggest the need for further studies to determine the potential medicinal and cosmetic applications (sunscreen) of lignin from C. tapia leaves.

Characterization and use of a lignin sample extracted from Eucalyptus grandis sawdust for the removal of methylene blue dye.

A lignin sample was extracted from Eucalyptus grandis sawdust, by the Klason method, and used as adsorbent for the removal of methylene blue (MB) from aqueous solutions. By using a set of complementary analytical tools, the lignin appeared to be constituted of oxygenated functional groups and aromatic moieties, while showing a specific surface area of 20 m2 g-1 and polydisperse particles. Different experimental conditions with various solid to liquid ratio, pH, as well as other external experimental parameters were investigated for the removal of MB by the lignin sample. The experimental adsorption data at the equilibrium were properly fitted by Langmuir model, while adsorption kinetical isotherms were correctly adjusted by the pseudo-second order model. The MB removal by lignin was spontaneous involving physisorption mechanisms leading to a saturation of the adsorption sites with a maximum adsorbed amount of about 32 mg g-1. The data acquired in this study also emphasized the interests to use lignin as potential adsorbent in the light of its properties for the removal of cationic dyes, including MB, with possible recycling and reuse cycles of lignin without any significant loss of its properties.

Valorization of orange bagasse through one-step physical and chemical combined processes to obtain a cellulose-rich material.

BACKGROUND: Orange bagasse (OB) is an agroindustrial residue of great economic importance that has been little explored for the extraction of cellulose. The present study aimed to investigate different combinations of chemical (sodium hydroxide, peracetic acid and alkaline peroxide) and physical (autoclaving and ultrasonication) treatments performed in one-step processes for cellulose extraction from OB and to characterize the materials obtained according to their composition, morphology, crystallinity and thermal stability. RESULTS: The processing yields ranged from 140 to 820 g kg-1 , with a recovery of 720-1000 g kg-1 of the original cellulose. Treatments promoted morphological changes in the fiber structure, resulting in materials with higher porosity, indicating partial removal of the noncellulosic fractions. The use of combined chemical treatments (NaOH and peracetic acid) with autoclaving was more efficient for obtaining samples with the highest cellulose contents. CONCLUSION: Therefore, ACSH (processed by autoclaving with NaOH) was the most effective one-step treatment, resulting in 71.1% cellulose, 0% hemicellulose and 19.0% lignin, with a crystallinity index of 42%. The one-step treatments were able to obtain materials with higher cellulose contents and yields, reducing reaction times and the quantity of chemical reagents employed in the overall processes compared to multistep conventional processes. © 2020 Society of Chemical Industry.

From cashew byproducts to biodegradable active materials: Bacterial cellulose-lignin-cellulose nanocrystal nanocomposite films.

While the cashew culture is focused on processing and commercialization of cashew nuts, the pseudofruit (cashew apples) - highly perishable and of limited acceptance - are mostly wasted. The cashew tree pruning fiber (CTPF) is another interesting cashew byproduct. In this study, films have been made from bacterial cellulose produced from cashew apple juice, and added with lignin (0-15 wt%) and cellulose nanocrystals (0-8 wt%), both from CTPF, which enhanced tensile properties and decreased water vapor permeability of the films. Moreover, lignin, although imparting brown color and opacity to the films, was effective to provide the films with UV-absorbing and antioxidant properties, making the films interesting for packaging of food products susceptible to lipid oxidation. The films did not exhibit antimicrobial activity against bacteria or yeasts.

Immunostimulatory and antioxidant activities of a lignin isolated from Conocarpus erectus leaves.

In this work, we investigated the antioxidant and immunomodulatory activities of a lignin isolated from Conocarpus erectus leaves. The lignin was characterized by FTIR, 1H NMR, 13C NMR and gel permeation chromatography analysis as well as ultraviolet/visible absorption spectra. The lignin was evaluated for total antioxidant activity (TAA), DPPH and ABTS+ scavenging abilities, and by a lipid peroxidation inhibition assay. Immunomodulatory activity of the lignin (10 µg/mL) on human peripheral blood mononuclear cells (PBMCs) was determined. The C. erectus lignin was found to be of the guaiacyl-syringyl-p-hydroxyphenyl (G-S-H) type, with an average molecular weight of 2709 Da (polydispersity index: 2.1). It showed low TAA (17.92%) and moderate antioxidant activity against DPPH and ABTS+ (IC50: 231.16 and 356.03 µg/mL, respectively). It also inhibited lipid peroxidation by 42.14%. The lignin promoted an increase in mitochondrial ROS levels as well as cytosolic Ca2+ in PBMCs. In addition, it promoted the differentiation and activation of CD8+ T lymphocytes, differentiation of CD14+ monocytes, and stimulated the release of nitric oxide and cytokines, mainly those linked to a Th1 response. The results showed that the C. erectus lignin may be used in future studies in which the modulation of the immune response is a key factor.

Enhancement of the Amazonian Açaí Waste Fibers through Variations of Alkali Pretreatment Parameters.

The açaí fruit depulping produces large amounts of long lignocellulosic fiber bundles that are disposed in the environment. Chemical pretreatments may improve açaí fibers favoring their usage in advanced materials. This work aimed to define optimal alkali reaction parameters to improve the properties of açaí fibers. Two NaOH concentrations (5 % and 10 %) and two reaction temperatures (80 °C and 100 °C) were tested. The raw and treated fibers were analyzed by scanning electron microscopy, Fourier transformed infrared spectroscopy, X-ray diffraction, and thermal analyses. All the alkali pretreatments separated fibers from the bundles, unblocked pit channels by removing silicon structures, exposed the inner lignin, partially removed non-cellulosic compounds, and raised the cellulose crystalline index. The highest temperature and NaOH content resulted in better cleaning and isolation of the fibers, while milder conditions better preserved the cellulose crystalline structure and thermal stability.

Isolation and physicochemical characterization of different lignin streams generated during the second-generation ethanol production process.

Four different lignin samples were isolated during the processing of rice straw for ethanol production: one after mild alkaline pretreatment (deacetylation) of rice straw, and other three after simultaneous saccharification and fermentation (fermentation residues), under different process conditions. Then, a complete chemical characterization in terms of main components (lignin, carbohydrates, among others) and elemental composition was performed for all the samples. Additionally, the structural and morphological characteristics, calorific value, thermal stability, solubility in solvents and antioxidant potential were also determined. For comparison, a standard lignin (Kraft lignin) was also assayed. The results revealed that the process used for lignin isolation has an important influence in the final properties of the sample. In general, the fermentation residues contained non-hydrolyzed polysaccharides in their composition and lower lignin content when compared to the lignin solubilized in the alkaline liquor. Among several organic solvents and mixtures, acetone 80% (v/v) was the most efficient for lignin solubilisation. Interesting properties and characteristics were observed for all the lignin samples isolated from rice straw processing, which would allow their application in different industrial areas. Based on the properties, potential applications were suggested, which could be of value for integration in biorefineries.

Lignins isolated from Prickly pear cladodes of the species Opuntia fícus-indica (Linnaeus) Miller and Opuntia cochenillifera (Linnaeus) Miller induces mice splenocytes activation, proliferation and cytokines production.

Opuntia fícus-indica and Opuntia cochenillifera are species of Cactaceae, found in the arid regions of the planet. They present water, cellulose, hemicellulose, pectins, extractives, ashes and lignins. Here we aimed to study the immunomodulatory action of lignins from these two species against mice splenocytes, since no study for this purpose has yet been reported. The antioxidant activities of these lignins were evaluated by the DPPH, ABTS, NO assays and total antioxidant activity. Cytotoxicity was evaluated through Annexin V-FITC and propidium iodide-PE probs and cell proliferation was determined by CFSE. Immunomodulation studies with Opuntia lignins obtained were performed through investigation of ROS levels, cytosolic calcium release, changes on mitochondrial membrane potential, cytokine production and NO release. Results showed that Opuntia cochenillifera lignin presented more phenolic amount and antioxidant activities than Opuntia ficius-indica. Both lignins showed high cell viability (>96%) and cell proliferation. Activation signal was observed for both lignins with increase of ROS and cytosolic calcium levels, and changes in mitochondrial membrane potential. In addition, lignins induced high TNF-α, IL-6 and IL-10 production and reduced NO release. Therefore, these lignins present great potential to be used as molecules with a proinflammatory profile, being shown as a promising therapeutic agent.

Comparison of lignin extraction processes: Economic and environmental assessment.

This paper presents the technical-economic and environmental assessment of four lignin extraction processes from two different raw materials (sugarcane bagasse and rice husks). The processes are divided into two categories, the first processes evaluates lignin extraction with prior acid hydrolysis step, while in the second case the extraction processes are evaluated standalone for a total analysis of 16 scenarios. Profitability indicators as the net present value (NPV) and environmental indicators as the potential environmental impact (PEI) are used through a process engineering approach to understand and select the best lignin extraction process. The results show that both economically and environmentally process with sulfites and soda from rice husk presents the best results; however the quality of lignin obtained with sulfites is not suitable for high value-added products. Then, the soda is an interesting option for the extraction of lignin if high quality lignin is required for high value-added products at low costs.

Use of agave bagasse for production of an organic fertilizer by pretreatment with Bjerkandera adusta and vermicomposting with Eisenia fetida.

Agave tequilana Weber is used in tequila and fructans production, with agave bagasse generated as a solid waste. The main use of bagasse is to produce compost in tequila factories with a long traditional composting that lasts 6-8 months. The aim of this study was to evaluate the degradation of agave bagasse by combining a pretreatment with fungi and vermicomposting. Experiments were carried out with fractionated or whole bagasse, sterilized or not, subjecting it to a pretreatment with Bjerkandera adusta alone or combined with native fungi, or only with native bagasse fungi (non-sterilized), for 45 days. This was followed by a vermicomposting with Eisenia fetida and sewage sludge, for another 45 days. Physicochemical parameters, lignocellulose degradation, stability and maturity changes were measured. The results indicated that up to 90% of the residual sugars in bagasse were eliminated after 30 days in all treatments. The highest degradation rate in pretreatment was observed in non-sterilized, fractionated bagasse with native fungi plus B. adusta (BNFns) (71% hemicellulose, 43% cellulose and 71% lignin) at 45 days. The highest total degradation rates after vermicomposting were in fractionated bagasse pre-treated with native fungi (94% hemicellulose, 86% cellulose and 91% lignin). However, the treatment BNFns showed better maturity and stability parameters compared to that reported for traditional composts. Thus, it seems that a process involving vermicomposting and pretreatment with B. adusta could reduce the degradation time of bagasse to 3 months, compared to the traditional composting process, which requires from 6 to 8 months.

The acetyl bromide method is faster, simpler and presents best recovery of lignin in different herbaceous tissues than Klason and thioglycolic acid methods.

We compared the amount of lignin as determined by the three most traditional methods for lignin measurement in three tissues (sugarcane bagasse, soybean roots and soybean seed coat) contrasting for lignin amount and composition. Although all methods presented high reproducibility, major inconsistencies among them were found. The amount of lignin determined by thioglycolic acid method was severely lower than that provided by the other methods (up to 95%) in all tissues analyzed. Klason method was quite similar to acetyl bromide in tissues containing higher amounts of lignin, but presented lower recovery of lignin in the less lignified tissue. To investigate the causes of the inconsistencies observed, we determined the monomer composition of all plant materials, but found no correlation. We found that the low recovery of lignin presented by the thioglycolic acid method were due losses of lignin in the residues disposed throughout the procedures. The production of furfurals by acetyl bromide method does not explain the differences observed. The acetyl bromide method is the simplest and fastest among the methods evaluated presenting similar or best recovery of lignin in all the tissues assessed.

Ethanol and lignin production from Brazilian empty fruit bunch biomass.

Brazil Government is promoting palm plantations to use degraded land for biofuels. Palm production is expected to increase 35 per cent in future and there would be profuse biomass available that needs to be handled efficiently. Therefore, in this study the potential of EFB from Brazil as raw material for biorefinery was explored by compositional analysis and pretreatment conditions optimization to produce ethanol and co-products. EFB from Brazil contains significant cellulose, hemicellulose, lignin and low ash content. The optimized dilute sulfuric acid pretreatment conditions for efficient cellulose and hemicellulose separation were 160°C temperature, 1.025% v/v acid concentration, 10.5min and 20% solid loading. Under optimum pretreatment process conditions, low enzyme loading (10FPU, 20IU cellulase and glucosidase enzyme/g glucan) and 15% solid loading, 51.1g ethanol, 344.1g solid residue (65% lignin and 24.87MJ/kg LHV) and 3.7l xylose rich liquid could be produced per kg dry EFB.

Evaluation of agave fiber delignification by means of microscopy techniques and image analysis.

Recently, the use of different types of natural fibers to produce paper and textiles from agave plants has been proposed. Agave atrovirens can be a good source of cellulose and lignin; nevertheless, the microstructural changes that happen during delignification have scarcely been studied. The aim of this work was to study the microstructural changes that occur during the delignification of agave fibers by means of microscopy techniques and image analysis. The fibers of A. atrovirens were obtained from leaves using convective drying, milling, and sieving. Fibers were processed using the Acetosolv pulping method at different concentrations of acetic acid; increasing acid concentration promoted higher levels of delignification, structural damage, and the breakdown of fiber clumps. Delignification followed by spectrometric analysis and microstructural studies were carried out by light, confocal laser scanning and scanning electron microscopy and showed that the delignification process follows three stages: initial, bulk, and residual. Microscopy techniques and image analysis were efficient tools for microstructural characterization during delignification of agave fibers, allowing quantitative evaluation of the process and the development of linear prediction models. The data obtained integrated numerical and microstructural information that could be valuable for the study of pulping of lignocellulosic materials.

Alkaline hydrogen peroxide pretreatment of cashew apple bagasse for ethanol production: study of parameters.

The alkaline hydrogen peroxide (AHP) pretreatment of cashew apple bagasse (CAB) was evaluated based on the conversion of the resultant cellulose into glucose. The effects of the concentration of hydrogen peroxide at pH 11.5, the biomass loading and the pretreatment duration performed at 35°C and 250 rpm were evaluated after the subsequent enzymatic saccharification of the pretreated biomass using a commercial cellulase enzyme. The CAB used in this study contained 20.56 ± 2.19% cellulose, 10.17 ± 0.89% hemicellulose and 35.26 ± 0.90% lignin. The pretreatment resulted in a reduced lignin content in the residual solids. Increasing the H2O2 concentration (0-4.3% v/v) resulted in a higher rate of enzymatic hydrolysis. Lower biomass loadings gave higher glucose yields. In addition, no measurable furfural and hydroxymethyl furfural were produced in the liquid fraction during the pretreatment. The results show that alkaline hydrogen peroxide is effective for the pretreatment of CAB.

Use of lignin extracted from different plant sources as standards in the spectrophotometric acetyl bromide lignin method.

A nongravimetric acetyl bromide lignin (ABL) method was evaluated to quantify lignin concentration in a variety of plant materials. The traditional approach to lignin quantification required extraction of lignin with acidic dioxane and its isolation from each plant sample to construct a standard curve via spectrophotometric analysis. Lignin concentration was then measured in pre-extracted plant cell walls. However, this presented a methodological complexity because extraction and isolation procedures are lengthy and tedious, particularly if there are many samples involved. This work was targeted to simplify lignin quantification. Our hypothesis was that any lignin, regardless of its botanical origin, could be used to construct a standard curve for the purpose of determining lignin concentration in a variety of plants. To test our hypothesis, lignins were isolated from a range of diverse plants and, along with three commercial lignins, standard curves were built and compared among them. Slopes and intercepts derived from these standard curves were close enough to allow utilization of a mean extinction coefficient in the regression equation to estimate lignin concentration in any plant, independent of its botanical origin. Lignin quantification by use of a common regression equation obviates the steps of lignin extraction, isolation, and standard curve construction, which substantially expedites the ABL method. Acetyl bromide lignin method is a fast, convenient analytical procedure that may routinely be used to quantify lignin.

Valorization of an industrial organosolv-sugarcane bagasse lignin: Characterization and use as a matrix in biobased composites reinforced with sisal fibers.

In the present study, the main focus was the characterization and application of the by-product lignin isolated through an industrial organosolv acid hydrolysis process from sugarcane bagasse, aiming at the production of bioethanol. The sugarcane lignin was characterized and used to prepare phenolic-type resins. The analysis confirmed that the industrial sugarcane lignin is of HGS type, with a high proportion of the less substituted aromatic ring p-hydroxyphenyl units, which favors further reaction with formaldehyde. The lignin-formaldehyde resins were used to produce biobased composites reinforced with different proportions of randomly distributed sisal fibers. The presence of lignin moieties in both the fiber and matrix increases their mutual affinity, as confirmed by SEM images, which showed good adhesion at the biocomposite fiber/matrix interface. This in turn allowed good load transference from the matrix to the fiber, leading to biobased composites with good impact strength (near 500 J m(-1) for a 40 wt% sisal fiber-reinforced composite). The study demonstrates that sugarcane bagasse lignin obtained from a bioethanol plant can be used without excessive purification in the preparation of lignocellulosic fiber-reinforced biobased composites displaying high mechanical properties.

Advanced oxidation of bleached eucalypt kraft pulp mill effluent.

In this study a poorly biodegradable (BOD/COD = 0.3) industrial alkaline ECF bleaching filtrate was treated using different advanced oxidation processes to evaluate their use in combined chemical-biological treatment aimed at increasing recalcitrant COD removal and improving final effluent quality. Oxidative treatments included ozonation combined with hydrogen peroxide (2, 5, 10, 20 mmol L(-1) O3/0.7, 2, 5, 10 mmol L(-1) H2O2) and photocatalysis with hydrogen peroxide (UV/2, 4 and 8 mmolL(-1) H2O2) and with TiO2 (UV/TiO2/0.7 and 4 mmol L(-1) H2O2). The O3/H2O2 process increased effluent biodegradability by up to 68% as a result of increasing BOD and decreasing COD. Increasing the O3 dose had a greater effect on biodegradability improvement and lignin and colour removal efficiencies than increasing the H2O2 dose. A combined oxidant dose of 5 mmol L(-1) O3 and 2 mmol L(-1) H2O2 resulted in 75% lignin removal, 40% colour removal and 6% carbohydrate loss without mineralizing the organic carbon. The photocatalytic processes led to a decrease in effluent biodegradability through combined decrease in BOD and increase in COD and did not result in efficient lignin or colour removal. Photocatalytic oxidation was apparently inhibited by the high chloride and COD levels in the alkaline filtrate, and may be more efficient in recalcitrant COD removal if performed after biological.

Combined chemical biological treatment of bleached eucalypt kraft pulp mill effluent.

Effectiveness of ozonation before and after biological treatment for removal of recalcitrant organic matter in bleached kraft pulp effluents was compared. Two industrial ECF bleached eucalypt kraft pulp effluents (E1 and E2) were pretreated with 100 mg O3/L. Raw and pretreated effluents were treated biologically in bench-scale sequencing batch reactors, under constant conditions. Following biological treatment, effluents were post-treated with 100 and 200 mg O3/L. Effluent pretreatment increased effluent biodegradability by 10% in E1 and 24% in E2. Combined O3-biological treated led to small but significant increases in COD, BOD and lignin removal over biological treatment alone, but pretreatment had no significant effect on effluent colour and carbohydrate removal. Ozone pretreatment did not affect biological activity during treatment of effluent E1 but resulted in a 38% lower specific oxygen uptake rate in effluent E2. At an equivalent dose of 100 mg/L, pre-ozonation produced better quality effluent than post-ozonation, especially with regard to COD and colour. Likewise, when an equivalent dose of 200 mg/L was applied, splitting the dose equally between pre- and post-treatments was more efficient than applying the entire dose in the post-treatment. The potential for combined chemical-biological treatment to improve effluent quality has been confirmed in this study.

Lignin concentration in oat (Avena byzantina L. ) aerial part as measured by four analytical methods

The current analytical methods to quantify lignin in forages are not satisfactory. A spectrophotometric method, the acetyl bromide lignin(ABL), has been employed to determine lignin concentration in forages; however, it suffers from the lack of an ideal standard with which the optical density readings of samples are compared to. A lignin, extracted from the plant with a solution of acidic dioxane, was employed to build a calibration curve for this method. This procedure was then compared with other methods (acid detergent lignin - ADL, Klason lignin – KL and potassium permanganate lignin – PerL) to determine lignin content on different fractions (stem, leaf and whole plant) of eight oat cultivars (Avena byzantina L.). There was no agreement among the four methods. In general, ABL and KL methods yielded thehighest values while ADL method yielded the lowest, particularly for the young plants. Lignin concentration was higher in the stem fractionas compared to leaf. It was detected influence of maturity stage in the investigated samples. It is concluded that the ABL method employingas standard lignin extracted with acidic dioxane has potential to be employed as a method to determine lignin concentration.

Os métodos analíticos para quantificar a concentração de lignina atualmente em uso não se tem mostrado satisfatórios. Um método espectrofotométrico, a lignina brometo de acetila (LBA) tem sido empregado para determinar o teor de lignina em plantas forrageiras; entretanto, padece da inexistência de um padrão de referência ideal, com o qual as leituras de densidade óptica das amostras são comparadas. Uma lignina, extraída da planta com solução ácida dedioxano, foi empregada para a construção de uma curva de calibração para o método em questão. Este procedimento foi comparado com outros métodos (lignina detergente ácido - LDA, lignina Klason –LK e lignina permanganato de potássio - LPer) na estimativa do teor de lignina em diferentes frações vegetais (caule, folha e parte aérea) de oito cultivares de aveia (Avena byzantina L.). Não houve concordância de valores entre os quatro métodos analíticos. Num âmbito geral,LBA e LK forneceram as maiores estimativas enquanto a LDA resultounos menores valores, particularmente nas amostras de plantas mais jovens. A concentração de lignina foi mais elevada na fração caule do que na folha. Foi detectado efeito da maturidade nas amostras analisadas. Conclui-se que o método LBA usando como padrão de referência a lignina extraída com dioxano ácido tem potencial para ser empregado nas determinações dos teores de lignina.