Production and Characterization of High Value Prebiotics From Biorefinery-Relevant Feedstocks.

Hemicellulose, a structural polysaccharide and often underutilized co-product stream of biorefineries, could be used to produce prebiotic ingredients with novel functionalities. Since hot water pre-extraction is a cost-effective strategy for integrated biorefineries to partially fractionate hemicellulose and improve feedstock quality and performance for downstream operations, the approach was applied to process switchgrass (SG), hybrid poplar (HP), and southern pine (SP) biomass at 160°C for 60 min. As a result, different hemicellulose-rich fractions were generated and the chemical characterization studies showed that they were composed of 76-91% of glucan, xylan, galactan, arabinan, and mannan oligosaccharides. The hot water extracts also contained minor concentrations of monomeric sugars (≤18%), phenolic components (≤1%), and other degradation products (≤3%), but were tested for probiotic activity without any purification. When subjected to batch fermentations by individual cultures of Lactobacillus casei, Bifidobacterium bifidum, and Bacteroides fragilis, the hemicellulosic hydrolysates elicited varied responses. SG hydrolysates induced the highest cell count in L. casei at 8.6 log10 cells/ml, whereas the highest cell counts for B. fragilis and B. bifidum were obtained with southern pine (5.8 log10 cells/ml) and HP hydrolysates (6.4 log10 cells/ml), respectively. The observed differences were attributed to the preferential consumption of mannooligosaccharides in SP hydrolysates by B. fragilis. Lactobacillus casei preferentially consumed xylooligosaccharides in the switchgrass and southern pine hydrolysates, whereas B. bifidum consumed galactose in the hybrid poplar hydrolysates. Thus, this study (1) reveals the potential to produce prebiotic ingredients from biorefinery-relevant lignocellulosic biomass, and (2) demonstrates how the chemical composition of hemicellulose-derived sources could regulate the viability and selective proliferation of probiotic microorganisms.

Structural changes in lignocellulosic biomass during activation with ionic liquids comprising 3-methylimidazolium cations and carboxylate anions.

BACKGROUND: Lignocellulosic biomass requires either pretreatment and/or fractionation to recover its individual components for further use as intermediate building blocks for producing fuels, chemicals, and products. Numerous ionic liquids (ILs) have been investigated for biomass pretreatment or fractionation due to their ability to activate lignocellulosic biomass, thereby reducing biomass recalcitrance with minimal impact on its structural components. In this work, we studied and compared 1-allyl-3-methylimidazolium formate ([AMIM][HCOO]) to the commonly used 1-ethyl-3-methylimidazolium acetate ([EMIM][CH3COO]) for its potential to activate hybrid poplar biomass and enable high cellulose and hemicellulose enzymatic conversion. Although [EMIM][CH3COO] has been widely used for activation, [AMIM][HCOO] was recently identified to achieve higher biomass solubility, with an increase of 40% over [EMIM][CH3COO]. RESULTS: Since IL activation is essentially an early stage of IL dissolution, we assessed the recalcitrance of [EMIM][CH3COO] and [AMIM][HCOO]-activated biomass through a suite of analytical tools. More specifically, Fourier transform infrared spectroscopy and X-ray diffraction showed that activation using [AMIM][HCOO] does not deacetylate hybrid poplar as readily as [EMIM][CH3COO] and preserves the crystallinity of the cellulose fraction, respectively. This was supported by scanning electron microscopy and enzymatic saccharification experiments in which [EMIM][CH3COO]-activated biomass yielded almost twice the cellulose and hemicellulose conversion as compared to [AMIM][HCOO]-activated biomass. CONCLUSION: We conclude that the IL [AMIM][HCOO] is better suited for biomass dissolution and direct product formation, whereas [EMIM][CH3COO] remains the better IL for biomass activation and fractionation.

Effects of Oligosaccharides Isolated From Pinewood Hot Water Pre-hydrolyzates on Recombinant Cellulases.

Loblolly pine residues have enormous potential to be the raw material for advanced biofuel production due to extensive sources and high cellulose content. Hot water (HW) pretreatment, while being a relatively economical and clean technology for the deconstruction of lignocellulosic biomass, could also inhibit the ensuing enzymatic hydrolysis process because of the production of inhibitors. In this study, we investigated the effect of oligosaccharide fractions purified from HW pre-hydrolyzate of pinewood using centrifugal partition chromatography (CPC) on three recombinant cellulolytic enzymes (E1, CBHI and CBHII), which were expressed in the transgenic corn grain system. The efficiency of recombinant enzymes was measured using either a 4-methylumbelliferyl-ß-D-cellobioside (MUC) or a cellulose-dinitrosalicylic acid (DNS) assay system. The results showed that HW pre-hydrolyzate CPC fractions contain phenolics, furans, and monomeric and oligomeric sugars. Among CPC fractions, oligomers composed of xylan, galactan, and mannan were inhibitory to the three recombinant enzymes and to the commercial cellulase cocktail, reducing the enzymatic efficiency to as low as 10%.

Sustainable Hydrogels Based on Lignin-Methacrylate Copolymers with Enhanced Water Retention and Tunable Material Properties.

Synthesizing lignin-based copolymers would valorize a major coproduct stream from pulp and paper mills and biorefineries as well as reduce the dependence on petrochemical-based consumer goods. In this study, we used organosolv lignin isolated from hybrid poplar ( Populus trichocarpa × P. deltoides) to generate lignin-containing methacrylate hydrogels. The copolymer hydrogels were synthesized by first grafting 2-hydroxyethyl methacrylate (HEMA) onto lignin (OSLH) via esterification and then by free radical polymerization of OSLH with excess HEMA. The copolymer hydrogels were prepared with different stoichiometric ratios of OSLH (e.g., 0, 10, 20, and 40 wt %) with respect to HEMA. Copolymerization with OSLH led to an increase in cross-linking density, which in turn enhanced the hydrogel's material properties; we report up to 39% improvement in water retention, 20% increase in thermostability, and up to a 3 order increase in magnitude of the storage modulus ( G'). The copolymer's properties, such as water retention and glass transition temperature, could be tuned by altering the percent functionalization of lignin OH groups and the ratio of OSLH to HEMA.

Applications of Trametes versicolor crude culture filtrates in detoxification of biomass pretreatment hydrolyzates.

Laccases have wide range of substrate specificity and find applications from pulp industry to waste water remediation. Laccases have also been used in combined pretreatment of biomass hydrolyzates to remove enzymatic and fermentation inhibitors. In this study, laccase production by Trametes versicolor strains isolated from different regions of the United States was induced using copper salts. T. versicolor crude culture filtrates (CCF), without any purification step, were tested for removal of model inhibitor compounds as well as in poplar and rice straw pretreatment hydrolyzates. Phenolic inhibitors were removed by 76% and 94% from the dilute acid hydrolyzates of rice straw and poplar, respectively, when incubated with the CCF for 12h, at room temperature. Xylo-oligosaccharide concentrations present in rice straw hydrolyzates were reduced by 64% when incubated with T. versicolor CCF. T. versicolor CCF could be a low cost technology for decreasing enzymatic and fermentation inhibitors.

Characterization of Rice Straw Prehydrolyzates and Their Effect on the Hydrolysis of Model Substrates Using a Commercial endo-Cellulase, ß-Glucosidase and Cellulase Cocktail.

Pretreatment and enzymatic saccharification are two major upstream processes that affect the economic feasibility and sustainability of lignocellulosic biofuel production. Cellulase-inhibiting degradation products, generated during dilute acid pretreatment, increase enzyme usage, and therefore, it is essential to mitigate their production. In an attempt to elucidate the most deleterious degradation product to enzymatic hydrolysis, hydrolyzates were generated from rice straw, and their effect on enzyme activity was determined. Ground rice straw was subjected to the following pretreatments having a combined severity factor of 1.75: T1-160 °C, pH 1.7; T2-180 °C, pH 2.25; and T3-220 °C, pH 7.0. The liquid prehydrolyzates were freeze-dried, and their inhibitory effects on the activities of a commercial cellulase cocktail, endo-cellulase, and ß-glucosidase were determined using filter paper, carboxymethyl cellulose, and cellobiose, respectively. Addition of 15 g L-1 of T1, T2, or T3 freeze-dried prehydrolyzates resulted in 67%, 57%, and 77% reduction in CMC-ase activity of endo-cellulase, respectively. In the presence of 35 g L-1 of T1, T2, or T3 prehydrolyzates, the filter paper activity of the cellulase cocktail was reduced by 64%, 68%, and 82%, respectively. Characterization of the freeze-dried prehydrolyzates showed that T3 had significantly higher xylo-oligosaccharides and total phenolic content than T2 and T1.

Separation of xylose oligomers using centrifugal partition chromatography with a butanol-methanol-water system.

Xylose oligomers are the intermediate products of xylan depolymerization into xylose monomers. An understanding of xylan depolymerization kinetics is important to improve the conversion of xylan into monomeric xylose and to minimize the formation of inhibitory products, thereby reducing ethanol production costs. The study of xylan depolymerization requires copious amount of xylose oligomers, which are expensive if acquired commercially. Our approach consisted of producing in-house oligomer material. To this end, birchwood xylan was used as the starting material and hydrolyzed in hot water at 200 °C for 60 min with a 4 % solids loading. The mixture of xylose oligomers was subsequently fractionated by a centrifugal partition chromatography (CPC) with a solvent system of butanol:methanol:water in a 5:1:4 volumetric ratio. Operating in an ascending mode, the butanol-rich upper phase (the mobile phase) eluted xylose oligomers from the water-rich stationary phase at a 4.89 mL/min flow rate for a total fractionation time of 300 min. The elution of xylose oligomers occurred between 110 and 280 min. The yields and purities of xylobiose (DP 2), xylotriose (DP 3), xylotetraose (DP 4), and xylopentaose (DP 5) were 21, 10, 14, and 15 mg/g xylan and 95, 90, 89, and 68 %, respectively. The purities of xylose oligomers from this solvent system were higher than those reported previously using tetrahydrofuran:dimethyl sulfoxide:water in a 6:1:3 volumetric ratio. Moreover, the butanol-based solvent system improved overall procedures by facilitating the evaporation of the solvents from the CPC fractions, rendering the purification process more efficient.

Hot water and dilute acid pretreatment of high and low specific gravity Populus deltoides clones.

Populus sp. are hardwood feedstocks that grow in forest management areas that are logged for softwoods; however, they are also being considered as an energy-destined feedstock. The objective of this work was to determine the effect of xylose yield from dilute acid and hot water pretreatments performed in unstirred batch stainless steel reactors at temperatures ranging from 140 to 200°C. Populus deltoides clones S13C20 and S7C15 used in this study originated from Eastern Texas and were cultivated for 14 years in Pine Tree, AR. P. deltoides clones S13C20 and S7C15 had specific gravities of 0.48 and 0.40, respectively. Bark and wood were examined separately. As expected, hot water pretreatments, in the tested temperature range, resulted in very little direct xylose recovery. However, the 140°C dilute acid pretreatment of the lower specific gravity clone, S7C15, wood yielded the highest average xylose recovery of 56%. This condition also yielded the highest concentration of furfural, 9 mg/g sample, which can be inhibitory to the fermentation step. The highest xylose recovery from bark samples, 31%, was obtained with clone S7C15, using the 160°C dilute acid pretreatment for 60 min.

Separation and purification of xylose oligomers using centrifugal partition chromatography.

Xylose oligomers, which have a prebiotic effect, have been used as additives to human and animal food. These oligomers are also the primary intermediate in hemicellulose degradation during the pretreatment of biomass. Centrifugal partition chromatography (CPC) was used in this study to separate and purify xylan-derived oligomers from birchwood xylan. The xylan was partially hydrolyzed to achieve varying degrees of polymerization at 130°C using 0.98% aqueous sulfuric acid for 20 min with a 2.5% solid loading. The CPC solvent system consisting of dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and water in a 1:6:3 volumetric ratio was used because of its ability to dissolve xylose oligomers of different degrees of polymerization. The CPC was operated in the ascending mode with the water- and DMSO-rich bottom phase acting as the stationary phase, while the THF-rich top phase was the eluent. This paper delineates a method for the production and purification of xylose monomer and xylose oligomers (up to xylopentaose) using CPC. The amount and purity of compounds collected from the CPC fractionation based on 1 g of birchwood xylan were 25.26 mg of xylose at 91.86% purity, 10.71 mg of xylobiose at 85.07% purity, 4.15 mg of xylotriose at 54.71% purity, 5.03 mg of xylotetraose at 38.33% purity and 3.31 mg of xylopentaose at 30.43% purity.

Sweetgum (Liquidambar styraciflua L.): extraction of shikimic acid coupled to dilute acid pretreatment.

Liquidambar styraciflua L., also known as sweetgum, is an understory hardwood species that has widespread distribution in the southeast USA, especially in pine plantations. In addition to being a possible biorefinery feedstock, sweetgum contains shikimic acid, which is a precursor for the drug Tamiflu. Sweetgum bark was extracted with 65 degrees C water and yielded 1.7 mg/g of shikimic acid, while sweetgum de-barked wood yielded 0.2 mg/g of shikimic acid. Because shikimic acid can be extracted with water, the coupling of the phytochemical hot water extraction with dilute acid pretreatment was examined. The addition of a 65 degrees C shikimic acid extraction step coupled to pretreatment with 0.98% H(2)SO(4) at 130 degrees C for 50 min resulted in 21% and 17% increases in xylose percent recovery from bark and de-barked wood, respectively. These results indicate that, in addition to recovering a high value product, the 65 degrees C wash step also increases xylose recovery.

Switchgrass water extracts: extraction, separation and biological activity of rutin and quercitrin.

Switchgrass (Panicum virgatum L.) has recently received significant attention as a possible feedstock for the production of liquid fuels such as ethanol. In addition, switchgrass may also be a source of valuable co-products, such as antioxidants, and our laboratory recently reported that switchgrass contains policosanols and alpha-tocopherol. Motivation for this work began when a switchgrass sample was extracted with water at 50 degrees C and was then tested for low-density lipoprotein (LDL) oxidation inhibition activity using the Thiobarbituric Acid Reactive Substances (TBARS) assay. The TBARS results showed that the switchgrass water extracts inhibited LDL oxidation by as much as 70% in comparison to the control. Liquid chromatography coupled with mass spectrometry (LC-MS) and high performance liquid chromatography (HPLC) were used to identify the compounds that were responsible for LDL oxidation inhibition activity as flavonoids: quercitrin (quercetin-3-O-rhamnoside) and rutin (quercetin-3-O-rutinoside). To maximize flavonoid concentrations, switchgrass was then extracted with water and 60% methanol at different temperatures. The 60% methanol treatment resulted in higher rutin and quercitrin yields when compared to water-only extraction; however, the use of this solvent would not be practical with current biorefinery technology. Centrifugal partition chromatography (CPC) was then used to purify rutin and quercitrin from the switchgrass water extract, which were then tested via the TBARS assay and shown to exhibit lipid peroxidation inhibition activity similar to that obtained with pure flavonoid standards. This is the first report on the presence of rutin and quercitrin in switchgrass. The results support the extraction of viable coproducts from switchgrass prior to conversion to liquid fuel.

Policosanol, alpha-tocopherol, and moisture content as a function of timing of harvest of Switchgrass (Panicum virgatum L.).

Using switchgrass ( Panicum virgatum L.) as a cellulosic feedstock for the production of ethanol could lead to the extraction of co-products prior to the pretreatment step, thereby adding value to the ethanol conversion process. Policosanols, registered as 142583-61-7, are present in Poaceae and are a mixture of long-chained primary alcohols. Policosanols are composed mainly of docosanol (C(22)), tetracosanol (C(24)), hexacosanol (C(26)), octacosanol (C(28)), triacontanol (C(30)), and dotriacontanol (C(32)). This study determined changes in moisture, policosanol, and alpha-tocopherol concentrations of Cave-in-Rock and Blackwell switchgrass cultivars during maturation from July to December in Arkansas and Oklahoma. Moisture content on a dry weight basis declined from 150 to 50% with progressive harvests. The total policosanol concentration ranged between 89 mg/kg for July harvested Cave-in-Rock switchgrass from Arkansas and 182 mg/kg for August harvested Cave-in-Rock switchgrass for Oklahoma, and these values remained relatively constant throughout the season. This is the first report on the presence of policosanols in switchgrass. Total switchgrass policosanol concentrations were lower than those typically reported for sorghum grains; however, switchgrass-extracted policosanols contained different policosanol ratios, wherein C(30) and C(32) alcohol ranges were 36-41 and 43-50%, respectively. alpha-Tocopherol extracted from both switchgrass cultivars varied between 320 and 400 mg/kg but decreased in the October harvest after frost.

Extraction of co-products from biomass: example of thermal degradation of silymarin compounds in subcritical water.

In an effort to increase revenues from a given feedstock, valuable co-products could be extracted prior to biochemical or thermochemical conversion with subcritical water. Although subcritical water shows significant promise in replacing organic solvents as an extraction solvent, compound degradation has been observed at elevated extraction temperatures. First order thermal degradation kinetics from a model system, silymarin extracted from Silybum marianum, in water at pH 5.1 and 100, 120, 140, and 160 degrees C were investigated. Water pressure was maintained slightly above its vapor pressure. Silymarin is a mixture of taxifolin, silichristin, silidianin, silibinin, and isosilibinin. The degradation rate constants ranged from 0.0104 min(-1) at 100 degrees C for silichristin to a maximum of 0.0840 min(-1) at 160 degrees C for silybin B. Half-lives, calculated from the rate constants, ranged from a low of 6.2 min at 160 degrees C to a high of 58.3 min at 100 degrees C, both for silichristin. The respective activation energies for the compounds ranged from 37.2 kJ/gmole for silidianin to 45.2 kJ/gmole for silichristin. In extracting the silymarin with pure ethanol at 140 degrees C, no degradation was observed. However, when extracting with ethanol/water mixtures at and 140 degrees C, degradation increased exponentially as the concentration of water increased.

Milk thistle extracts inhibit the oxidation of low-density lipoprotein (LDL) and subsequent scavenger receptor-dependent monocyte adhesion.

Silymarin encompasses a group of flavonolignans that are extracted from Silybum marianum (Asteraceae) fruits. The silymarins have previously been reported to lower low-density lipoprotein (LDL) levels associated with high-fat diets. The present study reports the efficacy of the silymarins in inhibiting oxidized low-density lipoprotein (oxLDL) generation and subsequent scavenger receptor (SR) mediated monocyte adherence to oxLDL. The flavonolignans that comprise silymarin include silichristin (SC), silidianin (SD), silibinin (SBN), and isosilibinin (IS). These flavonolignans (300 microM) lowered oxLDL generation, measured by the thiobarbituric acid-reacting substances (TBARS) assay, by 60.0, 28.1, 60.0, and 30.1%, respectively. SBN treatment of LDL in the presence of copper sulfate (CuSO 4) resulted in a significant dose-dependent inhibition of monocyte adhesion. Inhibition was paralleled by a decrease in binding of anti-oxLDL antibodies recognized by U937 monocyte Fc gamma receptors (FcgammaR). These results showed that silymarin and SBN, likely through antioxidant and free radical scavenging mechanisms of action, inhibit the generation of oxLDL and oxidation-specific neoepitopes recognized by SR and FcgammaR expressed on monocytes/macrophages.

Pretreatment of milk thistle seed to increase the silymarin yield: an alternative to petroleum ether defatting.

Milk thistle (Silybum marianum L.) seed meal is extracted for the flavonolignans, silychristin, silydianin, silybinin A, silybinin B, isosilybinin A and isosilybinin B, which are collectively known as the silymarin complex. To obtain the flavonolignans, the meal is usually treated with successive washes of petroleum ether to remove the lipids, followed by extraction of the flavonolignans with ethanol. This work examines the possible replacement of petroleum ether and ethanol by water or other aqueous solutions in these processes. To replace petroleum ether, pretreatments with 1.2% NaOH (w/w), 1.5% H2SO4 (w/w), 2% NaHCO3 (w/w), 0.14% cellulase and water were investigated. Of these pretreatments, 1.5% H2SO4 and water produced similar flavonolignan yields as petroleum ether. Results established that pretreating the milk thistle seed meal with 1.5% H2SO4 (w/w) at 50 degrees C for 18 h could replace the petroleum ether pretreatment. In addition, it was shown that similar amounts of flavonolignan could be recovered with a 1.5% H2SO4/water (100 degrees C) extraction as with a petroleum ether/ethanol extraction. Although cellulase pretreatment was not examined extensively, significant advances in cellulase effectiveness and cost have occurred in the past few years by companies such as Genencor International and Novozymes. These advances should help to make enzyme use for cellulose conversion, as well as extraction pretreatment, technically and economically feasible.

Pressurized water versus ethanol as a Silybum marianum extraction solvent for inhibition of low-density lipoprotein oxidation mediated by copper and J774 macrophage cells.

Silybum marianum contains flavonolignans, termed silymarin (SM), that are therapeutic agents for many inflammation-based diseases including atherosclerosis. Oxidation of human low-density lipoprotein was induced by CuSO4 or J774 macrophage cells and measured by the formation of thiobarbituric acid reactive substances (TBARS). SM was extracted by pressurized hot water (PHWE) or ethanol, and the effects of these extracts on TBARS formation were evaluated in comparison with those of SM preparations made from blending masses of individual flavonolignan standards in ratios identical to those of the water and ethanol extracts. Ethanol-extracted SM and its blended counterpart inhibited the generation of TBARS by 82% and 43%, respectively, at 150 mumol/L doses. TBARS levels in the presence of 150 micromol/L of the PHWE and its blended SM counterpart were reduced by 84% and 38%, respectively. Extracts from milk thistle fruit displayed higher protective effects than blended SM solutions of the same concentration with an identical compositional makeup. The appearance of degradation peaks in the water extract did not create any cytotoxic effects. Results of this study confirm that PHWE can be used to extract flavonolignans from milk thistle and that these extracts may possess therapeutic potential different from or beyond that of traditional organic solvent preparations.

Effect of Albizia julibrissin water extracts on low-density lipoprotein oxidization.

High-value phytochemicals could be extracted from biomass prior to the current cellulosic pretreatment technologies (i.e., lime, ammonia, dilute acid, or pressurized hot water treatments) provided that the extraction is performed with a solvent that is compatible with the pretreatment. This work reports on the extraction of flavonoids from Albizia julibrissin biomass. While extracting A. julibrissin foliage with 50 degrees C water, 2.227 mg/g of hyperoside and 8.134 mg/g quercitrin were obtained, which is in the realm of what was obtained with 60% methanol. A. julibrissin foliage, flower, and whole plant extracts were tested in terms of their potential to inhibit low-density lipoprotein (LDL) oxidization. The highest inhibition was obtained with foliage water extracts, which were standardized at 2.5 microM of flavonoids. Also, the 2.5 microM foliage water extract resulted in a reduction from 43% to only 1% of the observed monocyte adherence. To have commercial application, A. julibrissin water extracts should be devoid of toxicity. The A. julibrissin foliage, flower, and whole plant water extracts were not toxic to Vero 76 cells. In summary, A. julibrissin biomass can be extracted with 50 degrees C water to yield an antioxidant stream, showing that it may be possible to couple extraction of valuable phytochemicals to the cellulosic pretreatment step.

Extraction of hyperoside and quercitrin from mimosa (Albizia julibrissin) foliage.

Mimosa, an excellent energy crop candidate because of its high growth yield, also contains, on a dry basis, 0.83% hyperoside and 0.90% quercitrin. Hyperoside has been documented as having anti-inflammatory and diurectic properties, whereas quercitrin may play a role in intestinal repair following chronic mucosal injury. Thus, mimosa might first be extracted for important antioxidant compounds and then used as a feedstock for energy production. This article presents results from studies aimed at determining the effect of three extraction parameters (temperature, solvent composition, and time) on the yield of these important quercetin compounds. Conditions are sought which maximize yield and concentration, whereas complementing subsequent biomass pretreatment, hydrolysis and fermentation.

Silymarin extraction from milk thistle using hot water.

Hot water is attracting attention as an extraction solvent in the recovery of compounds from plant material as the search for milder and "greener" solvents intensifies. The use of hot water as an extraction solvent for milk thistle at temperatures above 100 degrees C was explored. The maximum extraction yield of each of the silymarin compounds and taxifolin did not increase with temperature, most likely because significant compound degradation occurred. However, the time required for the yields of the compounds to reach their maxima was reduced from 200 to 55 min when the extraction temperature was increased from 100 to 140 degrees C. Severe degradation of unprotected (plant matrix not present) silymarin compounds was observed and first-order degradation kinetics were obtained at 140 degrees C.

Extraction of antioxidant compounds from energy crops.

Energy crops offer enormous opportunities for increasing the sustain ability of agriculture and energy production in the United States. Nevertheless, opportunities for sustaining biomass energy production may well hinge on producing energy and extracting high-value products from the same crop. Seven potential energy crops (mimosa, sericea, kudzu, arunzo, switchgrass, velvet bean, and castor) were extracted and assayed for the presence of potentially high-value antioxidant compounds. Of these crops, mimosa and sericea had the highest antioxidant potential and were selected for further study. High-performance liquid chromatography (ultraviolet) and liquid chromatography/mass spectrometry techniques were then utilized to help identify the compounds with high antioxidant potential using extract fractionation, and total phenolics and oxygen radical absorbance capability assays as a guide. These analyses indicate that methanol extracts of mimosa foliage most likely contain quercetin, a flavonol that has been associated with cardio- protection. Future work will concentrate on quantifying the quercetin content of mimosa (likely parts-per-million levels), as well as identifying and quantifying other antioxidants found in energy crops.