Organic nitrogen in geomacromolecules: insights on speciation and transformation with K-edge XANES spectroscopy

Organic nitrogen incorporated into geomacromolecules (e.g., humic substances, kerogen) represents a major reservoir of nitrogen on the earth's surface, accounting for more than 90% of the total nitrogen in soils, sediments, and aquatic environments. Its primary source is biochemical nitrogen from dead plant and animal residues (predominantly proteinaceous substances), which undergo a complex series of transformations, mediated by microbes and abiotic processes, ultimately resulting in the incorporation of the nonmineralized fraction into geomacromolecules. Simultaneously,the biochemical N is thought to be extensively altered structurally, forming more stable structures (such as heterocyclic forms), although the type of changes in chemical speciation, their timing, and mechanisms are not clear. It is important to have this knowledge because the type of N formed influences not only its reactivity and fate (e.g., the release of bioavailable N in soils) but also the physical and chemical characteristics of the associated macromolecular organic matter. We used nitrogen K-edge XANES spectroscopy (a selective, sensitive, and nondestructive method) to gain new insights into the speciation of this macromolecular nitrogen. Our results verified amide N as being the dominant type in humic substances and sediments but revealed that pyridinic N also is a significant component of the total N (approximately 20-30%), with a subfraction consisting of its oxidized derivatives. An unidentified form of highly oxidized N was present mainly in sediments. While amide N represents residues of original biochemical molecules, pyridinic N probably is generated abiotically. Our results imply that the abiotic formation of pyridinic N sets in during the early stages of organic matter transformations thereby stabilizing organic N, although such processes generating heterocyclic structures may continue much longer.     

Solid- and solution-phase organics dictate copper distribution and speciation in multicomponent systems containing ferrihydrite, organic matter, and montmorillonite

Copper retention by ferrihydrite, leaf compost, and montmorillonite was studied over 8 months in systems that emulate a natural soil where different solid phases compete for Cu through a common solution in a compartmentalized batch reactor. Copper speciation in solution (total dissolved, DPASV-labile, and free) and exchangeable and total Cu in individual solid phases were determined. Organic carbon in solution (DOC) and that retained by the mineral phases were also determined. Cu sorption reached steady-state after 4 months and accounted for 80% of the Cu initially added to the system (0.15 mg L(-1)). The remaining 20% stayed in solution as nonlabile (82.8%), labile (17%), and free (0.2%) Cu species. Copper sorption followed the order organic matter > silicate clays > iron oxides. Within each solid phase, exchangeable Cu was < or = 10% of the total Cu sorbed. DOC reached steady state (22 mg L(-1)) after 4 months and seemed to control Cu solubility and sorption behavior by the formation of soluble Cu-DOC complexes and by sorbing onto the mineral phases. DOC sorption onto ferrihydrite prevented Cu retention by this solid phase. Using a multicomponent system and 8 months equilibrations, we were able to capture some of the more important aspects of the complexity of soil environments bytaking into account diffusion processes and competition among solid- and solution-phase soil constituents in the retention of a metal cation.     

Localization and speciation of chromium in subterranean clover using XRF, XANES, and EPR spectroscopy

Optimization of phytoremediation and assessment of potential health hazards from metals in the environment requires an understanding of absorption, localization, and transport of the target metal by plants. The objectives of this study were to localize Cr and determine the oxidation state and possible complexation mode of Cr in intact plant tissue by means of XANES, synchrotron XRF microprobe spectroscopy, and EPR spectroscopy. Subterranean clover (Trifolium brachycalycinum) was grown hydroponically with Cr(VI) (0.04-2.0 mmol L(-1)) and compared with plants grown without Cr and with inorganic Cr(III) and various Cr(III)-organic sources. The uptake, translocation, and form of Cr in the plant were dependent on the form and concentration of supplied Cr. Chromium was found predominately in the +3 oxidation state, regardless of the Cr source supplied to the plant, though at high Cr(VI) treatment concentrations, Cr(VI) and Cr(V) were also observed. At low Cr(VI) concentrations, the plant effectively reduced the toxic Cr(VI) to less toxic Cr(III), which was observed both as a Cr(III) hydroxide phase at the roots and as a Cr(III)-organic complex in the roots and shoots. At low Cr(VI) treatment concentrations, Cr in the leaves was observed predominately around the leaf margins, while at higher concentrations Cr was accumulated at leaf veins.     

Influence of natural organic matter source on copper speciation as demonstrated by Cu binding to fish gills, by ion selective electrode, and by DGT gel sampler

Rainbow trout (Oncorhynchus mykiss, 2 g) were exposed to 0-5 microM total copper in ion-poor water for 3 h in the presence or absence of 10 mg C/L of qualitatively different natural organic matter (NOM) derived from water spanning a large gradient in hydrologic residence time. Accumulation of Cu by trout gills was compared to Cu speciation determined by ion selective electrode (ISE) and by diffusive gradients in thin films (DGT) gel sampler technology. The presence of NOM decreased Cu uptake by trout gills as well as Cu concentrations determined by ISE and DGT. Furthermore, the source of NOM influenced Cu binding by trout gills with high-color, allochthonous NOM decreasing Cu accumulation by the gills more than low-color autochthonous NOM. The pattern of Cu binding to the NOM measured by Cu ISE and by Cu accumulation by DGT samplers was similar to the fish gill results. A simple Cu-gill binding model required an NOM Cu-binding factor (F) that depended on NOM quality to account for observed Cu accumulation by trout gills; values of Fvaried by a factor of 2. Thus, NOM metal-binding quality, as well as NOM quantity, are both important when assessing the bioavailability of metals such as Cu to aquatic organisms.     

X-ray absorption and micro X-ray fluorescence spectroscopy investigation of copper and zinc speciation in biosolids

Despite its pivotal role in determining the risks and time frames associated with contaminant release, metal speciation remains a poorly understood aspect of biosolids chemistry. The work reported here used synchrotron-based spectroscopy techniques to investigate the speciation of copper and zinc in a range of Australian biosolids. High resolution element mapping of biosolids samples using micro X-ray fluorescence spectroscopy revealed considerable heterogeneity in key element associations, and a combination of both organic and inorganic copper and zinc binding environments. Linear combination fitting of K-edge X-ray absorption spectra indicated consistent differences in metal speciation between freshly produced and stockpiled biosolids. While sulfide minerals play a dominant role in metal binding in freshly dewatered biosolids, they are of lesser importance in dried biosolids that have been stockpiled. A degree of metal binding with iron oxide minerals was apparent but the results did not support the hypothesis that biosolids metals are chiefly associated with iron minerals. This work has potential implications for the long-term stability of metals in biosolids and their eventual fate following land application.     

Mercury speciation by X-ray absorption fine structure spectroscopy and sequential chemical extractions: a comparison of speciation methods

Determining the chemical speciation of mercury in contaminated mining and industrial environments is essential for predicting its solubility, transport behavior, and potential bioavailability as well as for designing effective remediation strategies. In this study, two techniques for determining Hg speciation--X-ray absorption fine structure (XAFS) spectroscopy and sequential chemical extractions (SCE)--are independently applied to a set of samples with Hg concentrations ranging from 132 to 7539 mg/kg to determine if the two techniques provide comparable Hg speciation results. Generally, the proportions of insoluble HgS (cinnabar, metacinnabar) and HgSe identified by XAFS correlate well with the proportion of Hg removed in the aqua regia extraction demonstrated to remove HgS and HgSe. Statistically significant (>10%) differences are observed however in samples containing more soluble Hg-containing phases (HgCl2, HgO, Hg3S2O4). Such differences may be related to matrix, particle size, or crystallinity effects, which could affect the apparent solubility of Hg phases present. In more highly concentrated samples, microscopy techniques can help characterize the Hg-bearing species in complex multiphase natural samples.     

Hydrophobic interactions between spin-label 5-SASL and humic acid as revealed by ESR spectroscopy

The spin-label probe 5-SASL (stearic acid spin-label with nitroxide free radical in position 5 of hydrocarbon chain), detectable by electron spin resonance (ESR), was tested to evaluate pH and reaction time dependencies of hydrophobic interactions with humic acid (HA). Strong changes were observed in 5-SASL ESR spectra in the presence of HA suspensions below pH 5, with disappearance of the three isotropic narrow hyperfine lines of the nitroxide group (typical of free spin-label) and formation of "immobilized" 5-SASL spectra. These changes were interpreted as due to 5-SASL bonding with hydrophobic groups of HA, by van der Waals forces and/or hydrogen bonds, in very hydrophobic sites (probably water-protected) existent in HA below pH 5. However, such sites are absent above pH 5, as demonstrated by a specific experiment to check 5-SASL spectra reversibility. On the other hand, the HA suspension was more efficient in dissolving 5-SASL than water above pH 5. This fact also suggests the existence of "surface" hydrophobic sites, where the spin-label binds to HA while maintaining the nitroxide group in contact with water, as evidenced by the typical free spin-label spectrum and hyperfine interaction splitting (a0 = 1.574 mT). Also experiments checking 5-SASL reversibility bonding with HA were consistent with the supramolecular association model to HA.     

Fate of prions in soil: interactions of RecPrP with organic matter of soil aggregates as revealed by LTA-PAS

The contribution of soil organic matter (OM) to the adsorption of a recombinant prion protein (RecPrP) was studied in microcosm systems (soil aggregates from two different soils) before and after OM removal by low temperature ashing (LTA). The LTA technique allows a controlled removal of OM layer by layer, like a peeling of an onion skin, with minimal disturbance of the mineral matrix. Soil aggregates were selected as a representative model of the "in situ" conditions. Adsorption from batch vs percolation experiments were compared, and the aggregates were characterized by photoacustic Fourier-transform IR spectroscopy (PAS-FTIR). High affinity (H-type) adsorption isotherms were found with complete removal of RecPrP from solution for protein/soil ratios up to 1:62.5. OM removal from aggregates decreased the adsorbed RecPrP in amounts corresponding to 330-1000 microg mg(-1) of soil organic carbon (OC) indicating that native OM has specific adsorption capacity comparable and/or superior to the mineral matrix. The coupled LTA-PAS-FTIR approach demonstrated that, albeit OM composition was homogeneous throughout the aggregates, its presence in the most external surfaces of the aggregates affects the diffusion dynamics of RecPrP within the aggregates during percolation.     

Redox speciation of copper in rainwater: temporal variability and atmospheric deposition

Copper speciation was determined in 68 rainwater samples collected in Wilmington, NC, from August 25, 2000, to September 24, 2002. Volume-weighted average concentrations of Cu(total), dissolved Cu(II), and dissolved Cu(I) were 5.3, 3.2, and 1.4 nM, respectively, with a significantly higher ratio of Cu(II)/Cu(I) in summer relative to winter events. The concentrations of all Cu species were higher in storms of continental origin relative to marine-dominated events, suggesting anthropogenic and/or terrestrial sources are important contributors of Cu in precipitation. Concentrations of strong Cu-complexing ligands were consistently lower than dissolved Cu concentrations, indicating a significant portion, but not all, of the dissolved Cu in rainwater is strongly complexed. A portion of these ligands, in addition to the sulfite and chloride in precipitation, may be Cu(I)-complexing ligands, which may explain the resistance of Cu(I) against oxidation in rainwater. Using our rainwater concentration data along with other published rainwater Cu concentrations and an estimate for total global annual rain, the total global flux of Cu removed from the atmosphere via wet deposition is 150 x 106 kg yr(-1). This represents complete removal of the estimated Cu input into the troposphere and indicates essentially all Cu released into the global atmosphere is removed by rain.     

An improved method to identify irradiated rice by EPR spectroscopy and thermoluminescence measurements

Minimal change in irradiated foods with low dose treatment makes the identification process a difficult task. Two independent physical methods, electron paramagnetic resonance (EPR) spectroscopy and thermoluminescence (TL) detection were employed for detection of irradiation treatment on Basmati rice. EPR investigation of 0.5–2.0 kGy irradiated rice samples showed a short lived, asymmetric, dose dependent spectrum (g = 2.005), characterised by the radicals of irradiated starch. However, this signal disappears with time. The present work explores the possibility to identify irradiated rice by the relaxation characteristics and thermal behaviour of the radicals. This study reports for the first time that the different microwave saturation behaviours of the signal (g = 2.004) in irradiated and non-irradiated rice samples provide an important clue to identify radiation treatment beyond the period when the radiation specific EPR spectral lines have disappeared. TL investigation involving scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX) of the poly-minerals isolated from the rice samples allowed to discriminate clearly between irradiated and non-irradiated samples even after a prolonged period of storage.     

Carbon speciation of diesel exhaust and urban particulate matter NIST standard reference materials with C(1s) NEXAFS spectroscopy

The U.S. National Institute of Standards and Technology (NIST) provides a number of particulate matter (PM) standard reference materials (SRM) for use in environmental and toxicological methodology and research. We present here the first analysis with respect to the molecular structure of the carbon in three such NIST SRM samples, i.e., diesel engine exhaust soot from heavy duty equipment engines (SRM 1650), diesel soot from a forklift engine (SRM 2975), and urban PM collected in St. Louis, MO (SRM 1648), with near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. The NEXAFS spectra of the two diesel soot samples appear quite similar, while they differ significantly from the urban PM spectrum, in agreement with X-ray diffraction data published recently. Such comparison is made in terms of aromatic and aliphatic carbon species, as well as by a general comparison with graphitic materials. Both diesel soot SRM samples contain basic graphitic structures, but the presence of exciton resonance and extended X-ray absorption fine structure oscillations in SRM 1650 and the lack therof in SRM 2975 suggest that SRM 1650 is the more graphitic material.The presence of polycyclic aromatic hydrocarbons, which have a characteristic NEXAFS resonance at the same position as graphite, can obscure the graphitic character of soot, unless an extraction of the organic matter is made. Our NEXAFS data do not suggest that the urban PM sample SRM 1648 contains a substantial amount of graphite-like material.     

Identification of copper binding sites in soil organic matter through chemical modifications and 13C CP-MAS NMR spectroscopy

Metal binding to an organic peat soil was probed by paramagnetic doping with copper, chemical modifications of the organic matter in the soil, and 13C CP-MAS NMR spin lattice relaxation rate measurements. Carboxyl and hydroxyl functional groups were determined to be most significant in copper uptake by the unmodified soil. Esterification and acetylation of the soil showed that metal binding by carbohydrate structures occurs independently of other functional groups and may even induce a pseudochelation phenomenon. Sorption isotherms corroborate the importance of carbohydrate structures in metal binding. These results suggest that environmental modeling of metal binding and retention in soils should incorporate estimates of the distributions of all functional groups in the soil organic matter (e.g. aliphatic, carbohydrate, phenolic, carboxyl) and their relative binding strengths.     

丽春红S探针双波长吸收光谱法测定食品中的Cu

文中建立了测定食品中Cu的二元双波长可见吸收光谱(DWO-VIS)法。在pH 6.86的Tris-盐酸缓冲介质中,丽春红S与Cu(Ⅱ)发生反应,在可见光区生成具有一个明显正吸收峰和一个明显负吸收峰的离子缔合物。最大正吸收波长和最大负吸收波长分别为445 nm和510 nm。Cu(Ⅱ)在0.06~1.6 mg/L(正吸收,445 nm)、0.04~1.6 mg/L(负吸收,510 nm)和0.03~1.6 mg/L(DWO-VIS,445 nm+510 nm)内遵从朗伯-比尔定律,表观摩尔吸光系数(κ)分别为8.70×10~3(正吸收法)、2.04×10~4(负吸收法)和2.92×10~4(DWO-VIS法)L/(mol·cm),定量限分别为0.62、0.38和0.26 mg/100 g。还探讨了吸收光谱特征、适宜反应条件及共存物质的影响。双波长法用于饼干、面条及奶粉中Cu的测定,加标回收率为97.9%~102%,相对标准偏差RSD(n=5)为2.5%~2.8%。     

Sulfur speciation in soil by S K-Edge XANES spectroscopy: comparison of spectral deconvolution and linear combination fitting

Defined, quartz-diluted mixtures of sulfur (S) compounds with different oxidation state (OS) were analyzed by K-edge XANES spectroscopy using linear combination fitting (LCF) and spectrum deconvolution by fitting several Gaussian and arctangent functions (GCF). Additionally, for different soils the S speciation as calculated by both methods was compared with results of a wet-chemical S speciation. For mixtures of FeS, L-cysteine, and Na2SO4, the S speciation was recovered with satisfactory accuracy and precision by both methods at the 2 and 0.2 mg S g(-1) level. For GCF, white-line peaks must be normalized with respect to their OS-specific absorption cross-section. LCF must be conducted with dilute reference compounds to avoid self-absorption effects. For mixtures of FeS, FeS2, S°, and L-cysteine, both procedures showed poor accuracy. For the soils, similar percentages of reduced inorganic S, organic S, and sulfate were calculated by LCF, GCF, and wet chemical S speciation. GCF allows a fair estimation of S species groups with different OS (inorganic reduced S, organic reduced S, organic intermediate S, oxidized S) in soils without standards. If dilute standards of all S compounds assumed to be present in a sample are available, LCF is more objective and allows a more detailed S speciation.     

Protein and water structural changes in fish surimi during gelation as revealed by isotopic H/D exchange and Raman spectroscopy

Structural changes of proteins and water during gelation of fish surimi, have been studied by isotopic H/D exchange of water and Raman spectroscopy assisted by monitoring of rheological characteristics, in order to get insights into the structural and functional properties of surimi gels. The results indicate the following: (i) Protein hydrogen bond rearrangements occur involving mainly α-helix to β-sheet transition, (ii) the relative intensity of the symmetric H₂O stretching band near 3220 cm⁻¹ tends to decrease upon gelation, (iii) H/D exchange reveal a slower deuteration kinetics in the gels as compared to the surimi, (iv) the low temperature scanning electron microscopy shows a smaller pore size of the gel network as compared to the surimi, and suggests that water domains in gel are more inaccessible to D₂O, which is consistent with higher water holding capacity in the gel.     

Surface complexation of copper(II) on soil particles: EPR and XAFS studies

The interactions of transition metals with natural systems play an important role in the mobility and the bioavailability of these metals in soils. In this study, the adsorption of copper(II) onto natural soil particles was studied as a function of pH and metal concentration. The retention capacity of soil particles was determined at pH 6.2 to be equal to 6.7 mg of copper/g of solid. The Langmuir and Freundlich isotherm equations were then used to describe the partitioning behavior of the system at different pH values. A combination of EPR, extended X-ray absorption fine structure (EXAFS), and X-ray absorption near-edge structure (XANES) spectroscopies was used to probe the Cu atomic environment at the soil particles/aqueous interface. The spectroscopic study revealed that copper(II) ions are held in inner-sphere surface complexes. It also revealed that Cu was in an octahedral coordination with first-shell oxygen atoms. A weak tetragonal distortion was pointed out due to the Jahn-Teller effect, with a mean Cu-Oequatorial bond distance of 1.96 A and a Cu-Oaxial bond distance of 2.06 A. A detailed analysis of the spectroscopic data suggested that Cu(II) was bonded to organic matter coated onto the mineral fraction of soil particles.     

Detection of irradiation treatment in dried mushrooms by photostimulated luminescence,EPR spectroscopy and thermoluminescence measurements

Detection of the irradiation treatment of nine species of dried mushrooms commercially available employing photostimulated luminescence (PSL), electron paramagnetic resonance spectroscopy (EPR/ESR) and thermoluminescence (TL) is described. PSL is adaptable to six to seven species shortly after irradiation, but the PSL signal decays with storage and may render detection impossible. After 4 months of storage, six species could still be well identified by PSL if the radiation dose was higher than 5 kGy. EPR spectroscopy enabled detection with five to six species, both cellulose and crystalline sugar-like radicals being observed. For some mushrooms, hat or leg parts showed a different response and some lots could not be identified at all. TL - although being more cumbersome - allowed unequivocal identification of all species of investigated mushrooms. It is concluded that dried mushrooms exposed to ionising radiation can be successfully detected by at least one of the analytical methods investigated.     

Evidence for bioavailable copper-dissolved organic matter complexes and transiently increased copper bioavailability in manure-amended soils as determined by bioluminescent bacterial biosensors

The short-term (3 months) dynamics of bioavailable copper (Cu) species was determined in soils amended with various amounts of manure and Cu. Bioavailable Cu species were operationally defined as those species that were able to induce gene expression in a Cu-specific Pseudomonas fluorescens biosensor. Biosensor measurements were backed by analysis of total Cu in soil and of total Cu and free Cu2+ ion activity in solution. Cu bioavailability relative to the total Cu concentration increased dramatically with increasing Cu loading of manure and with increasing manure amendment to soil. In both cases, the immediate increase in bioavailability could be explained in part by increased Cu concentration in solution and in part by an increased bioavailability of dissolved Cu species. In contrast to Cu bioavailability, Cu2+ ion activity decreased progressively with increasing manure loading. Cu bioavailability declined rapidly during the weeks after manure amendment concomitant with a marked slow-down of C mineralization indicating a shift from initially bioavailable Cu-dissolved organic matter (Cu-DOM) complexes to nonavailable Cu-DOM complexes over time. Our data do not support the conventional view of metal bioavailability being primarily related to the free metal ion activity and strongly suggest differential bioavailability of Cu-DOM complexes in manure-amended soils.     

Size-based speciation of natural colloidal particles by flow field flow fractionation, inductively coupled plasma-mass spectroscopy, and transmission electron microscopy/X-ray energy dispersive spectroscopy: colloids-trace element interaction

Flow field flow fractionation (FIFFF), inductively coupled plasma-mass spectroscopy (ICP-MS), and transmission electron microscopy (TEM) coupled to X-ray energy dispersive spectrometry (X-EDS) are used in series for the first time to characterize colloids. Results demonstrate the utility of FIFFF-ICP-MS-TEM/X-EDS to relate physical properties (size) of colloids to their chemical properties (chemical composition, surface chemical composition, and colloids-trace elements association). Results suggest that the major part of natural organic matter (NOM) is concentrated in the fraction < 0.01 microm (C2). Aluminum, iron, and manganese are the main colloidal components in the fraction 0.01-0.45 microm (C1). Aluminum occurs as aluminum oxides or aluminosilicates in the whole size range, while iron and manganese occur as individual oxyhydroxides in the size range < 0.20 microm. Within the C2 fraction, Al, Mn, Cu, and Ni elements are complexed to NOM (e.g., humic substances). Iron is complexed to NOM in some samples and probably free in other samples. Lead is totally free in all samples. Within the C1 fraction, Cu and Pb are mostly associated to Fe and Mn oxyhydroxides. Consequently, NOM with Fe and Mn oxyhydroxides are the main colloidal carriers of trace elements in the Loire watershed system.