Lignin plays a vital role in the economics of biorefineries, serving as a source of process energy and a feedstock for sustainable fuels and chemical production. While understanding lignin's chemical composition is crucial, emerging evidence suggests that a more comprehensive understanding of its macromolecular structure is critical to explaining its complex behavior in the biorefinery. This study investigated the collapse of the lignin network in corn stover feedstock after harvest and storage as a result of the microbial digestion of hemicellulose. Fluorescence microscopy was used to detect the collapse of lignin by the changes in lignin's fluorescence lifetime, anisotropy, and the number of effective emitters. Our in situ microscopic results revealed lignin's coil-globule transition phenomena, which was only previously predicted by molecular dynamics modeling of extracted lignin in solvent. This collapse of lignin macromolecular structure was supported by results from NMR, IR, Raman, and powder X-ray diffraction. Our study revealed that the two major approaches for lignin valorization in the lignin-first biorefinery model, namely, monomer extraction and milled wood lignin extraction, were negatively impacted by the lignin collapse. As changes during storage are a source of feedstock variability, our study highlights the importance of understanding the effect of feedstock handling on biorefinery operations and economics.

Woody biomass represents an abundant resource for sustainable biofuels, biochemicals, and bioproducts. Technologies for converting woody biomass have been established for decades, and research consistently highlights the critical role of inorganic species and ash plays in feedstock handling and conversion processes, including equipment plugging, corrosion, and catalyst deactivation. A thorough understanding of the variability, transport behavior, and downstream impact of inorganic species in woody biomass is essential for defining feedstock quality specifications and developing effective management strategies for conversion processes. This review compiles critical information in five main sections: 1) inorganic species concentration in woody biomass, based on anatomical fractions and their sources of variability; 2) technique features for quantifying inorganic elemental chemical analysis; 3) impacts of inorganic species on biomass preprocessing; 4) impacts of inorganic species on pyrolysis, and 5) mitigation strategies. Additionally, this review explores future challenges and opportunities in addressing the impacts of inorganic species on biomass quality. These insights aim to support the sustainable development of the biomass-to-bioenergy pipeline and ensure high-quality lignocellulosic feedstocks for efficient downstream conversions. The findings offer valuable guidance to policy makers, industry stakeholders, and researchers in developing effective strategies for managing inorganic species in woody biomass and fostering the sustainable processes for lignocellulosic biorefineries.

The impacts of biological degradation on surface area, surface energy, wettability, and cohesion of anatomically fractionated (i.e., leaf, stalk, and cob) and bulk corn stover are presented in this study. The physical, thermal and chemical properties of corn stover are critical material attributes that not only influence the mechanical processing and chemical conversion of corn stover, but also the bulk solids handling and transport. The measured surface areas were observed to be dependent on the degree of biological degradation (mild vs. moderate vs. severe) and on the anatomical fraction. The surface area of the bulk corn stover samples increased with the degree of biological degradation. The leaf fraction was the most sensitive to biological degradation, resulting in an increase in surface area from 0.5 m2/g (mildly degraded) to 1.2 m2/g (severely degraded). In contrast, the surface area of the cob fraction remained relatively unaffected by the degree of biological degradation (i.e., mildly degraded−0.55 m2/g, severely degraded−0.40 m2/g. All biologically degraded samples resulted in significant changes to the surface chemistry (evidenced by an increase in surface energy. As a general trend, the surface energy of bulk corn stover increased with the degree of biological degradation—the same trend was observed for the leaf and stalk anatomical fractions; however, the surface energy for the cob fraction remained unchanged. Wettability, calculated from surface energy, for bulk corn stover samples did not reveal any discernable trend with the degree of biological degradation. However, trends in wettability were observed for the anatomical fractions, with wettability increasing for the stalk and leaf fractions, and decreasing for the cob fraction. Excluding the cob fraction, the work of cohesion increased with the degree of biological degradation. Understanding the impacts of biological degradation on the physical, chemical and thermal properties of corn stover offers insights to improve the overall operational reliability, efficiency and economics of integrated biorefineries.

Imaging in the visible spectrum is a low-cost tool that can be readily deployed for in-field or over-belt monitoring of biomass quality for bio-refining operations. Rapid image analysis coupled with innovative preprocessing may reduce the impacts of feedstock variability through identification of contaminants or other material attributes to guide selective sorting and quality management. Image analysis was employed to evaluate the quality of corn stover in red-green-blue (RGB) chromatic space. This study used controlled, bench-scale imaging as a proof-of-concept for rapid quality assessment of corn stover based on variations in material attributes, including chemical and physical attributes, that relate to biological degradation and soil contamination. Logistic regression-based classification algorithms were used to develop a method for biomass screening as a function of biological degradation or soil contamination. This study demonstrated the use of image analysis to extract features from RGB color space to investigate variations in critical material attributes from chemical composition of corn stover. Fourier transform infrared (FT-IR) suggested a correlation between red band intensity and biological degradation, while detailed surface texture analysis was found to distinguish among variations in ash. These insights offer promise for development of a rapid screening tool that could be deployed by farmers for in-field assessment of biomass quality or biorefinery operators for in-line sorting and process optimization.

An atomic force microscope (AFM) fundamentally measures the interaction between a nanoscale AFM probe tip and the sample surface. If the force applied by the probe tip and its contact area with the sample can be quantified, it is possible to determine the nanoscale mechanical properties (e.g., elastic or Young's modulus) of the surface being probed. A detailed procedure for performing quantitative AFM cantilever-based nanoindentation experiments is provided here, with representative examples of how the technique can be applied to determine the elastic moduli of a wide variety of sample types, ranging from kPa to GPa. These include live mesenchymal stem cells (MSCs) and nuclei in physiological buffer, resin-embedded dehydrated loblolly pine cross-sections, and Bakken shales of varying composition. Additionally, AFM cantilever-based nanoindentation is used to probe the rupture strength (i.e., breakthrough force) of phospholipid bilayers. Important practical considerations such as method choice and development, probe selection and calibration, region of interest identification, sample heterogeneity, feature size and aspect ratio, tip wear, surface roughness, and data analysis and measurement statistics are discussed to aid proper implementation of the technique. Finally, co-localization of AFM-derived nanomechanical maps with electron microscopy techniques that provide additional information regarding elemental composition is demonstrated.

An atomic force microscope (AFM) fundamentally measures the interaction between a nanoscale AFM probe tip and the sample surface. If the force applied by the probe tip and its contact area with the sample can be quantified, it is possible to determine the nanoscale mechanical properties (e.g., elastic or Young's modulus) of the surface being probed. A detailed procedure for performing quantitative AFM cantilever-based nanoindentation experiments is provided here, with representative examples of how the technique can be applied to determine the elastic moduli of a wide variety of sample types, ranging from kPa to GPa. These include live mesenchymal stem cells (MSCs) and nuclei in physiological buffer, resin-embedded dehydrated loblolly pine cross-sections, and Bakken shales of varying composition. Additionally, AFM cantilever-based nanoindentation is used to probe the rupture strength (i.e., breakthrough force) of phospholipid bilayers. Important practical considerations such as method choice and development, probe selection and calibration, region of interest identification, sample heterogeneity, feature size and aspect ratio, tip wear, surface roughness, and data analysis and measurement statistics are discussed to aid proper implementation of the technique. Finally, co-localization of AFM-derived nanomechanical maps with electron microscopy techniques that provide additional information regarding elemental composition is demonstrated.

Biomass quality is influenced by water's abundance, distribution, and status in relation to other chemical species within the polymer matrix. Water interacts with polymers that make up the cell walls, and these interactions govern the physical and chemical changes that occur during the storage and preprocessing of biomass feedstocks. Time-domain nuclear magnetic resonance (TD-NMR) was employed to explore variations in the physical constraints of water within the lignocellulosic microstructure in distinct anatomical fractions of biomass and as a function of biological degradation. The Carr-Purcell-Meiboom-Gill sequence, when combined with knowledge of the chemical composition and physical structure of pine residues and corn stover anatomical fractions, gives an accurate measurement of the bound and free water. In this work, the impacts of storage and biological degradation were investigated to elucidate changes in the status and distribution of water within distinct plant tissues. We also investigate how degradation during storage affects water interactions in different pine residues (e.g., bark, branch, and needle) and corn stover (e.g., cob, leaf, and stalk) anatomical fractions using transverse relaxation times (T-2). As demonstrated herein, TD-NMR provides quantitative data on lignocellulosic biomass-water interactions within anatomical fractions, which can further aid in the investigation of preprocessing effects on feedstock quality. Our findings suggest that biological heating enhances biomass-water interactions at the cellular and macromolecular scale. In addition, analysis of three-dimensional scanning electron microscopy reconstructions indicates that surface roughness wavelengths align with microscale roughness, suggesting that pine forestry residue and corn stover particles have primarily hydrophobic exterior surfaces. This study offers multiscale insights into understanding the microstructure, wettability, and chemical environment that dictate diffusion, enzyme access, and recalcitrance of lignocellulosic biomass.

The variability of chemical, physical, and mechanical properties of lignocellulosic biomass feedstocks has a major impact on the efficiency of biomass processing and conversion to fuels and chemicals. Storage conditions represent a key source of variability that may contribute to biomass quality variations from the time of harvest until delivery to the biorefinery. In some cases, substantial microbial degradation can take place during storage. In this work, we investigate how degradation during storage affects the surface texture, surface energy, and porosity of different corn stover anatomical fractions (e.g., leaf, stalk, and cob). Understanding any potential changes in surface properties is important because interparticle interactions during bioprocessing cause aggregation and blockages that lead to at least process inefficiency and at most complete equipment failure. The surface roughness and texture parameters of corn stover with variable degrees of microbial degradation were calculated directly from stereomicroscopy and scanning electron microscopy micrographs. Surface energy and porosity were measured by inverse gas chromatography. The results show differing trends in the impact of increasing biological heating and degradation depending on the specific corn stover tissue type that was analyzed. These results also indicate that biomass surface properties are scale-dependent and that the scale, which is most industrially relevant, may depend on the specific unit operation within the biorefinery being considered.

A comprehensive review of the literature shows that enzyme hydrolysis efficiency decreases with increased solids loadings at constant enzyme:cellulose ratios for pretreated lignocellulosic substrates. In seeking a mechanistic explanation for this phenomenon, we found that a nitrogen atmosphere enhances enzyme hydrolysis and minimizes the decrease in glucose yields as solids loadings are increased in an agitated bioreactor. For liquid hot water pretreated corn stover, at solids loadings of both 100 and 200 g/L and hydrolyzed for 72 hr in a 1 L bioreactor at pH 5.0 with 3.6 mg protein per g biomass, glucose yields were 55% in a nitrogen atmosphere versus 45% in air with agitation and about 34% without agitation. While mixing promotes biomass/enzyme contact and disperses sugars released during hydrolysis that would otherwise cause product inhibition, nitrogen gas displaces air, avoiding deactivation of cellulases by oxygen. The nitrogen effect points to a facile approach of enhancing hydrolysis at high solids loadings.

Bioenergy is an important renewable energy option worldwide, but the industry is susceptible to a myriad of risks including biomass supply, of which drought plays a role. Crops yields decrease during drought, increasing year-to-year risk for the agricultural industry. For the renewable energy industry, in particular, the effect of drought on crops is substantial and complex. This chapter discusses the current state of knowledge regarding how drought affects biomass destined for renewable energy as it relates to dry biomass yields and chemistry, the latter of which heavily impacts cost of production and final product yields. Advanced supply systems are one option for reducing biomass supply risk. These systems lead to higher, less variable crop yields during uncontrollable events like drought; however, the quality of material supplied in a drought year may still vary as drought impacts plant chemistry. This chapter provides analysis for chemical composition of four bioenergy crops observing that both carbohydrates and lignin decrease during a drought year compared to a year with minimal to no drought. These chemical changes can impact biochemical conversion through inhibitor formation and altering degradability during pretreatment.

The impacts and variability of inorganic material (measured as total ash) on surface area, surface energy, wettability, and cohesion of corn stover samples from Iowa were examined. The impact of total ash on the acid component of surface energy was determined to be significant with the acid component increasing with increasing ash content, in particular, the samples with total ash contents greater than 10%. Negligible effects with ash content were observed on both the dispersive and base components of surface energy. The work of cohesion for the compositions of corn stover with varying amounts of total inorganic matter increased with increasing total ash content, giving rise to potential bulk solids handling and transport challenges related to segregation, agglomeration, rat-holing, arching, and discontinuous flow patterns. Both the wettability (hydrophilicity) and work of cohesion increased for compositions of corn stover with increasing total inorganic content. Washing proved effective at removing extrinsic inorganic material from corn stover with a reduction in total ash content from 20.4% to 6.2%, accompanied by a significant reduction (from 85.6 to 42.5 mJ/m(2)) in the acid component of the surface energy.

Traditional approaches for corn stover utilization create an inhomogeneous, whole plant mixture that does not achieve performance and cost targets for biofuel production. The improvement of conversion performance will require one to gain a fundamental understanding of the inherent cellular structure and ash content in the process. This study investigated cellular structure and inorganic species contents with scanning electron microscopy (SEM), advanced energy dispersive X-ray spectroscopy (EDS), and wet analytical chemistry methods. These techniques provided a complementary means to understand variability within the dynamic cell walls and the distributions of inorganic species at the anatomical and tissue scales for various corn stover fractions. Material properties were investigated for bales from in-field storage scenarios ranging from mild to severe biological degradation. Results show that stem, cob, and leaf tissues all maintained cell wall and surface structure integrity after mild biological degradation. However, severe degradation (with spontaneous heating) caused remarkable structural distortion and cell wall shrinkage for all tissue types. Inorganic species quantitation data shows that three fractions of severe degradation have slightly reduced total ash, 4.91% vs 3.92% for cobs, 7.28% vs 5.98% for stems, and 14.21% vs 13.07% for leaves. Silica contents also decreased (2.67% vs 1.83% for cobs, 3.60% vs 2.89% for stems, and 9.72% vs 8.48% for leaves), likely due to the loss of extrinsic inorganic species. SEM-EDS inorganic species mapping of stems demonstrated that severe biological degradation caused the translocation of the dominant silicon elements from pith to rind tissue as well as a reduction of potassium in the stem. These findings offer detailed knowledge of feedstock variability at the anatomical tissue level that can be used to inform the development of advanced processing strategies. Innovative storage and selective fractionation processes have the potential to manage low-quality fractions/tissues and remove unfavorable inorganic species for efficient conversion of corn stover as well as other lignocellulosic biomass.

Feedstock variability that originates from biomass production and field conditions propagates through the value chain, posing a significant challenge to the emerging biorefinery industry. Variability in feedstock properties impacts feeding, handling, equipment operations, and conversion performance. Feedstock quality attributes, and their variations, are often overlooked in assessing feedstock value and utilization for conversion to fuels, chemicals, and products. This study developed and employed a multiscale analytical characterization approach coupled with data analytic methods to better understand the sources and distribution of feedstock quality variability through evaluation of 24 corn stover bales collected in 4 counties of Iowa. In total, 216 core samples were generated by sampling nine positions on each bale using a reliable bale coring process. The samples were characterized for a broad suite of physicochemical properties ranging across field and bale, macro, micro, and molecular scales. Results demonstrated that feedstock quality attributes can vary at all spatial scales and that multiple sources of variability must be considered in order to establish and manage biomass quality for conversion processes.

Industrial hemp has gained resurgent interest recently for applications in various sectors. This study evaluated 11 different industrial hemp [6 fiber-only and 5 dual-purpose (fiber and grain)] cul...

Production of biofuels, bioproducts, and bioenergy requires a well-characterized, stable, and reasonably uniform biomass supply and well-established supply chains for shipping biomass from farm fields to biorefineries, while achieving year-round production targets. Preserving and stabilizing biomass feedstock during storage is a necessity for cost-effective and sustainable biofuel production. Ensiling is a common storage method used to preserve and even improve forage quality; however, the impact of ensiling on biomass physical and chemical properties that influence bioconversion processes has been variable. Our objective in this work was to determine the effects of ensiling on lignocellulosic feedstock physicochemical properties and how that influences bioconversion requirements. We observed statistically significant decreases (p < 0.05) in the content of two major structural carbohydrates (glucan and xylan) of 5 and 8%, respectively, between the ensiled and non-ensiled materials. We were unable to detect differences in sugar yields from structural carbohydrates after pretreatment and enzymatic hydrolysis of the ensiled materials compared to non-ensiled controls. Based on this work, we conclude that ensiling the corn stover did not change the bioconversion requirements compared to the control samples and incurred losses of structural carbohydrates. At the light microscopy level, ensiled corn stover exhibited little structural change or relocation of cell wall components as detected by immunocytochemistry. However, more subtle structural changes were revealed by electron microscopy, as ensiled cell walls exhibit ultrastructural characteristics such as wall delimitation intermediate between non-ensiled and dilute-acid-pretreated cell walls. These findings suggest that alternative methods of conversion, such as deacetylation and mechanical refining, could take advantage of lamellar defects and may be more effective than dilute acid or hot water pretreatment for biomass conversion of ensiled materials.

Feedstock variability is a significant barrier to the scale-up and commercialization of lignocellulosic biofuel technologies. Variability in feedstock characteristics and behavior creates numerous challenges to the biorefining industry by affecting continuous operation and bioffiels yields. Currently, feedstock variability is understood and explained largely on the basis of chemical composition. Physical and mechanical properties and behavior of lignocellulosic feedstock in various unit operations, studied through advanced analytical methods, can further explain variability. Such studies will enable us in developing processes and designing equipment to improve operation and conversion performance. In this perspective, we review several advanced analytical methods that measure density, moisture content, thermal properties, flowability, grindability, rheology properties, and micromorphological characteristics. We also discuss the correlations and interactions among these properties that reflect the complexity of lignocellulosic biomass as a feedstock and the associated quality metrics and logistics of supplying consistent quality feedstock to a biorefinery. We also examine methods that have not traditionally been used to characterize lignocellulosic feedstocks but have the potential to bridge the gap in our explanation of feedstock variability.

Almost 36million tons of paper and yard trimmings are not currently being recycled or composted annually and likely end up in landfills. One approach to valorize these resources would be for biofuels production. Nonrecyclable municipal solid waste (MSW) paper and yard trimmings were assessed for their suitability as feedstocks to produce biofuels from sugars. Both materials met the cost target of less than $85.51/ton ($2016) with MSW paper ranging from $36.40 to $62.18/dry ton and grass clippings from $25.64 to $45.86/dry ton. MSW paper exceeded the carbohydrate requirement of 59% while grass clippings had significantly lower amounts of carbohydrates (30%). However, both types of MSW had ash contents significantly above the required 5% total ash with paper at 22% ash and grass clippings at 16%. While neither waste material was individually found to be suitable for biofuels production based on compositional requirements, it was determined that blending either waste with corn stover would produce materials that met carbohydrate, ash, and cost targets. The higher carbohydrate and lower ash stover compensated for the lower carbohydrates and high ash of the waste materials, while the lower cost waste materials offset the higher cost of corn stover.

This report summarizes the findings of a qualitative analysis to identify integrated strategies needed for more affordable biofuels. It outlines five key strategies needed to achieve lower fuel production costs in an integrated biorefinery and provides high-level research needs across the biofuel supply chain.