Accumulation and depuration of trinitrotoluene and related extractable and nonextractable (bound) residues in marine fish and mussels.

To determine if trinitrotoluene (TNT) forms nonextractable residues in mussels (Mytilus galloprovincialis) and fish (Cyprinodon variegatus) and to measure the relative degree of accumulation as compared to extractable TNT and its major metabolites, organisms were exposed to water fortified with (14)C-TNT. After 24 h, nonextractable residues made up 75% (mussel) and 83% (fish) while TNT accounted for 2% of total radioactivity. Depuration half-lives for extractable TNT, aminodinitrotoluenes (ADNTs) and diaminonitrotoluenes (DANTs) were fast initially (<0.5 h), but slower for nonextractable residues. Nonextractable residues from organisms were identified as ADNTs and DANTs using 0.1 M HCL for solubilization followed by liquid chromatography-tandem mass spectrometry. Recovered metabolites only accounted for a small fraction of the bound residue quantified using a radiotracer likely because of low extraction or hydrolysis efficiency or alternative pathways of incorporation of radiolabel into tissue.

Effect of varying feedstock-pretreatment chemistry combinations on the formation and accumulation of potentially inhibitory degradation products in biomass hydrolysates.

A variety of potentially inhibitory degradation products are produced during pretreatment of lignocellulosic biomass. Qualitative and quantitative interrogation of pretreatment hydrolysates is paramount to identifying potential correlations between pretreatment chemistries and microbial inhibition in downstream bioconversion processes. In the present study, corn stover, poplar, and pine feedstocks were pretreated under eight different chemical conditions, which are representative of leading pretreatment processes. Pretreatment processes included: 0.7% H(2)SO(4), 0.07% H(2)SO(4), liquid hot water, neutral buffer solution, aqueous ammonia, lime, lime with oxygen pressurization, and wet oxidation. Forty lignocellulosic degradation products resulting from pretreatment were analyzed using high performance liquid chromatography in combination with UV spectroscopy or tandem mass spectrometry detection (HPLC-PDA-MS/MS) and ion chromatography (IC). Of these compounds, several have been reported to be inhibitory, including furfural, hydroxymethyl furfural, ferulic acid, 3,4-dihydroxybenzaldehyde, syringic acid among others. Formation and accumulation of monitored compounds in hydrolysates is demonstrated to be a function of both the feedstock and pretreatment conditions utilized.

Compositional analysis of water-soluble materials in switchgrass.

Any valuation of a potential feedstock for bioprocessing is inherently dependent upon detailed knowledge of its chemical composition. Accepted analytical procedures for compositional analysis of biomass water-soluble extracts currently enable near-quantitative mass closure on a dry weight basis. Techniques developed in conjunction with a previous analytical assessment of corn stover have been applied to assess the composition of water-soluble materials in four representative switchgrass samples. To date, analytical characterization of water-soluble material in switchgrass has resulted in >78% mass closures for all four switchgrass samples, three of which have a mass closure of >85%. Over 30 previously unknown constituents in aqueous extracts of switchgrass were identified and quantified using a variety of chromatographic techniques. Carbohydrates (primarily sucrose, glucose, and fructose) were found to be the predominant water-soluble components of switchgrass, accounting for 18-27% of the dry weight of extractives. Total glycans (monomeric and oligomeric sugars) contributed 25-32% to the dry weight of extractives. Additional constituents contributing to the mass balance for extractives included various alditols (2-3%), organic acids (10-13%), inorganic ions (11-13%), and a distribution of oligomers presumed to represent a diverse mixture of lignin-carbohydrate complexes (30-35%). Switchgrass results are compared with previous analyses of corn stover extracts and presented in the context of their potential impact on biomass processing, feedstock storage, and future analyses of feedstock composition.

Compositional analysis of water-soluble materials in corn stover.

Corn stover is one of the leading feedstock candidates for commodity-scale biomass-to-ethanol processing. The composition of water-soluble materials in corn stover has been determined with greater than 90% mass closure in four of five representative samples. The mass percentage of water-soluble materials in tested stover samples varied from 14 to 27% on a dry weight basis. Over 30 previously unknown constituents of aqueous extracts were identified and quantified using a variety of chromatographic techniques. Monomeric sugars (primarily glucose and fructose) were found to be the predominant water-soluble components of corn stover, accounting for 30-46% of the dry weight of extractives (4-12% of the dry weight of feedstocks). Additional constituents contributing to the mass balance for extractives included various alditols (3-7%), aliphatic acids (7-21%), inorganic ions (10-18%), oligomeric sugars (4-12%), and a distribution of oligomers tentatively identified as being derived from phenolic glycosides (10-18%).

Pseudo reaction kinetics of organic degradation products in dilute-acid-catalyzed corn stover pretreatment hydrolysates.

A variety of degradation products are produced upon pretreatment of lignocellulosic biomass with dilute acid. To date, the complexity of these samples has significantly limited the scope of efforts to perform summative analyses of degradation products. Qualitative and quantitative interrogation of hydrolysates is also paramount to identifying potential correlations between pretreatment chemistry and microbial inhibition in downstream bioconversion processes. A recently developed reversed-phase high performance liquid chromatography technique with UV detection has been applied to perform quantitative assessments of a variety of hydrolysate components as a function of pretreatment time and temperature. Correlations of product concentrations to the pretreatment severity function indicate differing responses of various compounds to the kinetic influences of temperature and reaction time. Of the compounds measured, four demonstrated initial accumulation rates were sufficiently linear over the time period tested to enable determination of activation energy E(a). All four compounds appear to demonstrate higher E(a) than that assumed in the commonly applied severity function. Overall accumulation trends for most compounds indicated similar under-estimation of apparent activation energy by the severity function. Biotechnol. Bioeng. 2007;98: 1135-1145. (c) 2007 Wiley Periodicals, Inc.

Effect of dissolved carbon dioxide on accumulation of organic acids in liquid hot water pretreated biomass hydrolyzates.

Liquid hot water pretreatment has been proposed as a possible means of improving rates of enzymatic hydrolysis of biomass while maintaining low levels of inhibitory compounds. Supplementation of liquid hot water pretreatment with dissolved carbon dioxide, yielding carbonic acid, has been shown to improve hydrolysis of some biomass substrates compared with the use of water alone. Previous studies on the application of carbonic acid to biomass pretreatment have noted a higher pH of hydrolyzates treated with carbonic acid as compared with the samples prepared with water alone. This study has applied recently developed analytical methods to quantify the concentration of organic acids in liquid hot water pretreated hydrolyzates, prepared with and without the addition of carbonic acid. It was observed that the addition of carbon dioxide to liquid hot water pretreatment significantly changed the accumulated concentrations of most measured compounds. However, the measured differences in product concentrations resulting from addition of carbonic acid did not account for the measured differences in hydrolyzate pH.

High-performance liquid chromatography method for simultaneous determination of aliphatic acid, aromatic acid and neutral degradation products in biomass pretreatment hydrolysates.

A variety of degradation products are produced upon dilute acid pretreatment of lignocellulosic biomass. Within this larger construct, organic acids, phenols and aromatic aldehydes represent important compound classes to investigate due to increasing evidence of their inhibitory effect on fermentative microorganisms. An analytical extraction procedure is presented, enabling isolation of potential analytes away from alternative products in biomass hydrolysates. Additionally, a reversed-phase high-performance liquid chromatography (HPLC) method has been developed and validated, affording simultaneous separation and quantitative determination of 32 potential analytes in water with UV detection at 210 nm. The method was subsequently employed to quantify a variety of aliphatic acid, aromatic acid, aldehyde and phenolic degradation products in a corn-stover hydrolysate at concentration levels ranging from 0.02 to 41 mM.

(E)-1-(2-Hydroxy-4-methoxyphenyl)-2-(3,4,5-trimethoxyphenyl)ethene.

In the crystal structure of the title compound, C(18)H(20)O(5), all geometric parameters fall within experimental error of the expected values. Analysis of the molecular-packing plots reveals an infinite one-dimensional linear array running parallel to the c axis, formed by an O-H...O intermolecular hydrogen-bonding interaction. The stilbene framework and most of the substituents are approximately coplanar.