Biogenic formation and growth of uraninite (UO₂)

Biogenic UO? (uraninite) nanocrystals may be formed as a product of a microbial reduction process in uranium-enriched environments near the Earth's surface. We investigated the size, nanometer-scale structure, and aggregation state of UO? formed by iron-reducing bacterium, Shewanella putrefaciens CN32, from a uranium-rich solution. Characterization of biogenic UO? precipitates by high-resolution transmission electron microscopy (HRTEM) revealed that the UO? nanoparticles formed were highly aggregated by organic polymers. Nearly all of the nanocrystals were networked in more or less 100 nm diameter spherical aggregates that displayed some concentric UO? accumulation with heterogeneity. Interestingly, pure UO? nanocrystals were piled on one another at several positions via UO?-UO? interactions, which seem to be intimately related to a specific step in the process of growing large single crystals. In the process, calcium that was easily complexed with aqueous uranium(VI) appeared not to be combined with bioreduced uranium(IV), probably due to its lower binding energy. However, when phosphate was added to the system, calcium was found to be easily associated with uranium(IV), forming a new uranium phase, ningyoite. These results will extend the limited knowledge of microbial uraniferous mineralization and may provide new insights into the fate of aqueous uranium complexes.     

Quantitative separation of monomeric U(IV) from UO2 in products of U(VI) reduction

The reduction of soluble hexavalent uranium to tetravalent uranium can be catalyzed by bacteria and minerals. The end-product of this reduction is often the mineral uraninite, which was long assumed to be the only product of U(VI) reduction. However, recent studies report the formation of other species including an adsorbed U(IV) species, operationally referred to as monomeric U(IV). The discovery of monomeric U(IV) is important because the species is likely to be more labile and more susceptible to reoxidation than uraninite. Because there is a need to distinguish between these two U(IV) species, we propose here a wet chemical method of differentiating monomeric U(IV) from uraninite in environmental samples. To calibrate the method, U(IV) was extracted from known mixtures of uraninite and monomeric U(IV) and tested using X-ray absorption spectroscopy (XAS). Monomeric U(IV) was efficiently removed from biomass and Fe(II)-bearing phases by bicarbonate extraction, without affecting uraninite stability. After confirming that the method effectively separates monomeric U(IV) and uraninite, it is further evaluated for a system containing those reduced U species and adsorbed U(VI). The method provides a rapid complement, and in some cases alternative, to XAS analyses for quantifying monomeric U(IV), uraninite, and adsorbed U(VI) species in environmental samples.     

Evidence of uranium and associated trace element mobilization and retention processes at Oklo (Gabon), a naturally radioactive site

The processes that affect the mobility of uranium and other radionuclides in the environment have been largely studied at both the laboratory and the field scales. The natural reactors found at the Oklo uranium mine in Gabon constitute a unique investigation setting as spontaneous fission reactions occurred two billion years ago. Oklo uraninites contain a large amount of other radionuclides as a result of the fission process. We have investigated the dissolution behavior of four uraninite samples from Oklo as a function of temperature (25 and 60 degrees C) and bicarbonate concentration (2.7-30 mmol/L). We have also investigated the dissolution behavior of minor components of the uraninites (i.e., Nd, Cs, Mo, Yb, and Sb) in relation to the dissolution of uranium. The results of the reported work are in good agreement with the kinetic rate laws derived from other uranium(IV) dioxide studies. Some of the minor components are found to be congruently released from the uraninite phase, while it is postulated that dissolution from segregated phases might affect the final concentrations of some of the rare earth elements, i.e., Nd and Yb. In addition, we have performed dissolution studies at 60 degrees C with two uraninites representative of different geochemical environments at Oklo, to study the uranium dissolution rates as a function of the temperature. This has allowed derivation of apparent activation energies for the bicarbonate-promoted oxidative dissolution of the Oklo uraninites. The dissolution behavior of the minor components of the uraninites at 60 degrees C was found to closely follow the behavior observed at 25 degrees C. This indicates that similar codissolution mechanisms operate in the temperature range studied. The implications for the mobility of uranium and other radionuclides in natural and anthropogenic environments are discussed.     

Non-uraninite products of microbial U(VI) reduction

A promising remediation approach to mitigate subsurface uranium contamination is the stimulation of indigenous bacteria to reduce mobile U(VI) to sparingly soluble U(IV). The product of microbial uranium reduction is often reported as the mineral uraninite. Here, we show that the end products of uranium reduction by several environmentally relevant bacteria (Gram-positive and Gram-negative) and their spores include a variety of U(IV) species other than uraninite. U(IV) products were prepared in chemically variable media and characterized using transmission electron microscopy (TEM) and X-ray absorption spectroscopy (XAS) to elucidate the factors favoring/inhibiting uraninite formation and to constrain molecular structure/composition of the non-uraninite reduction products. Molecular complexes of U(IV) were found to be bound to biomass, most likely through P-containing ligands. Minor U(IV)-orthophosphates such as ningyoite [CaU(PO(4))(2)], U(2)O(PO(4))(2), and U(2)(PO(4))(P(3)O(10)) were observed in addition to uraninite. Although factors controlling the predominance of these species are complex, the presence of various solutes was found to generally inhibit uraninite formation. These results suggest a new paradigm for U(IV) in the subsurface, i.e., that non-uraninite U(IV) products may be found more commonly than anticipated. These findings are relevant for bioremediation strategies and underscore the need for characterizing the stability of non-uraninite U(IV) species in natural settings.     

Application of high-angle annular dark field scanning transmission electron microscopy,scanning transmission electron microscopy-energy dispersive X-ray spectrometry,and energy-filtered transmission electron microscopy to the characterization of nanoparticles in the environment

A major challenge to the development of a fundamental understanding of transport and retardation mechanisms of trace metal contaminants (<10 ppm) is their identification and characterization at the nanoscale. Atomic-scale techniques, such as conventional transmission electron microscopy, although powerful, are limited by the extremely small amounts of material that are examined. However, recent advances in electron microscopy provide a number of new analytical techniques that expand its application in environmental studies, particularly those concerning heavy metals on airborne particulates or water-borne colloids. High-angle annular dark field scanning transmission electron microscopy (HAADF-STEM), STEM-energy-dispersive X-ray spectrometry (EDX), and energy-filtered TEM (EFTEM) can be effectively used to identify and characterize nanoparticles. The image contrast in HAADF-STEM is strongly correlated to the atomic mass: heavier elements contribute to brighter contrast. Gold nanocrystals in pyrite and uranium nanocrystals in atmospheric aerosols have been identified by HAADF-STEM and STEM-EDX mapping and subsequently characterized by high-resolution TEM (HRTEM). EFTEM was used to identify U and Fe nanocrystals embedded in an aluminosilicate. A rare, As-bearing nanophase, westerveldite (FeAs), was identified by STEM-EDX and HRTEM. The combined use of these techniques greatly expands the effective application of electron microscopy in environmental studies, especially when applied to metals of very low concentrations. This paper describes examples of how these electron microbeam techniques can be used in combination to characterize a low concentration of heavy metals (a few ppm) on nanoscale particles.     

Thermodynamic constraints on the oxidation of biogenic UO2 by Fe(III) (Hydr)oxides

Uranium mobility in the environment is partially controlled by its oxidation state, where it exists as either U(VI) or U(IV). In aerobic environments, uranium is generally found in the hexavalent form, is quite soluble, and readily forms complexes with carbonate and calcium. Under anaerobic conditions, common metal respiring bacteria can reduce soluble U(VI) species to sparingly soluble UO2 (uraninite); stimulation of these bacteria, in fact, is being explored as an in situ uranium remediation technique. However, the stability of biologically precipitated uraninite within soils and sediments is not well characterized. Here we demonstrate that uraninite oxidation by Fe(III) (hydr)oxides is thermodynamically favorable under limited geochemical conditions. Our analysis reveals that goethite and hematite have a limited capacity to oxidize UO2(biogenic) while ferrihydrite can lead to UO2(biogenic) oxidation. The extent of UO2(biogenic) oxidation by ferrihydrite increases with increasing bicarbonate and calcium concentration, but decreases with elevated Fe(II)(aq) and U(VI)(aq) concentrations. Thus, our results demonstrate that the oxidation of UO2(biogenic) by Fe(III) (hydr)oxides may transpire under mildly reducing conditions when ferrihydrite is present.     

Oxidative Dissolution of Biogenic Uraninite in Groundwater at Old Rifle, CO

Reductive bioremediation is currently being explored as a possible strategy for uranium-contaminated aquifers such as the Old Rifle site (Colorado). The stability of U(IV) phases under oxidizing conditions is key to the performance of this procedure. An in situ method was developed to study oxidative dissolution of biogenic uraninite (UO?), a desirable U(VI) bioreduction product, in the Old Rifle, CO, aquifer under different variable oxygen conditions. Overall uranium loss rates were 50-100 times slower than laboratory rates. After accounting for molecular diffusion through the sample holders, a reactive transport model using laboratory dissolution rates was able to predict overall uranium loss. The presence of biomass further retarded diffusion and oxidation rates. These results confirm the importance of diffusion in controlling in-aquifer U(IV) oxidation rates. Upon retrieval, uraninite was found to be free of U(VI), indicating dissolution occurred via oxidation and removal of surface atoms. Interaction of groundwater solutes such as Ca2? or silicate with uraninite surfaces also may retard in-aquifer U loss rates. These results indicate that the prolonged stability of U(IV) species in aquifers is strongly influenced by permeability, the presence of bacterial cells and cell exudates, and groundwater geochemistry.     

Uranium(VI) reduction by iron(II) monosulfide mackinawite

Reaction of aqueous uranium(VI) with iron(II) monosulfide mackinawite in an O(2) and CO(2) free model system was studied by batch uptake measurements, equilibrium modeling, and L(III) edge U X-ray absorption spectroscopy (XAS). Batch uptake measurements showed that U(VI) removal was almost complete over the wide pH range between 5 and 11 at the initial U(VI) concentration of 5 × 10(-5) M. Extraction by a carbonate/bicarbonate solution indicated that most of the U(VI) removed from solution was reduced to nonextractable U(IV). Equilibrium modeling using Visual MINTEQ suggested that U was in equilibrium with uraninite under the experimental conditions. X-ray absorption near edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) spectroscopy showed that the U(IV) phase associated with mackinawite was uraninite. Oxidation experiments with dissolved O(2) were performed by injecting air into the sealed reaction bottles containing mackinawite samples reacted with U(VI). Dissolved U measurement and XAS confirmed that the uraninite formed from the U(VI) reduction by mackinawite did not oxidize or dissolve under the experimental conditions. This study shows that redox reactions between U(VI) and mackinawite may occur to a significant extent, implying an important role of the ferrous sulfide mineral in the redox cycling of U under sulfate reducing conditions. This study also shows that the presence of mackinawite protects uraninite from oxidation by dissolved O(2). The findings of this study suggest that uraninite formation by abiotic reduction by the iron sulfide mineral under low temperature conditions is an important process in the redistribution and sequestration of U in the subsurface environments at U contaminated sites.     

Redox behavior of uranium at the nanoporous aluminum oxide-water interface: implications for uranium remediation

Sorption-desorption experiments show that the majority (ca. 80-90%) of U(VI) presorbed to mesoporous and nanoporous alumina could not be released by extended (2 week) extraction with 50 mM NaHCO(3) in contrast with non-nanoporous α alumina. The extent of reduction of U(VI) presorbed to aluminum oxides was semiquantitatively estimated by comparing the percentages of uranium desorbed by anoxic sodium bicarbonate between AH(2)DS-reacted and unreacted control samples. X-ray absorption spectroscopy confirmed that U(VI) presorbed to non-nanoporous alumina was rapidly and completely reduced to nanoparticulate uraninite by AH(2)DS, whereas reduction of U(VI) presorbed to nanoporous alumina was slow and incomplete (<5% reduction after 1 week). The observed nanopore size-dependent redox behavior of U has important implications in developing efficient remediation techniques for the subsurface uranium contamination because the efficiency of in situ bioremediation depends on how effectively and rapidly U(VI) bound to sediment or soil can be converted to an immobile phase.     

Electron microbeam investigation of uranium-contaminated soils from Oak Ridge, TN, USA

Two samples of uranium-contaminated soil from the Department of Energy's Oak Ridge Reservation in Oak Ridge, Tennessee were investigated using electron microprobe analysis and transmission electron microscopy. The objectives of this research were to identify and characterize soil particles and rock chips with high uranium concentrations, to investigate the extent of uranium penetration into chips of parent material, and to identify solid-phase hosts for uranium in the samples. Three distinct solid-phase hosts for uranium have been identified: (1) iron oxyhydroxides, including goethite and ferrihydrite; (2) mixed Mn-Fe oxides; and (3) discrete uranium phosphates. In all three, uranium is associated with phosphorus. The ubiquitous U-P association highlights the influence of phosphate on the environmental fate of uranium. Uranium-bearing phases are found well within chips of weathered shale, as far as 900 microm from fractures and chip edges, indicating that uranium has diffused into the shale matrix.     

Reoxidation of reduced uranium with iron(III) (hydr)oxides under sulfate-reducing conditions

In cultures of Desulfovibrio desulfuricans 620 the effects of iron(III) (hydr)oxides (hematite, goethite, and ferrihydrite) on microbial reduction and reoxidation of uranium (U) were evaluated under lactate-limited sulfate-reducing conditions. With lactate present, G20 reduced U(VI) in both 1,4-piperazinediethanesulfonate (PIPES) and bicarbonate buffer. Once lactate was depleted, however, microbially reduced U served as an electron donor to reduce Fe(III) present in iron(III) (hydr)oxides. With the same initial amount of Fe(III) (10 mmol/L) for each iron(III) (hydr)oxide, reoxidation of U(IV) was greater with hematite than with goethite orferrihydrite. As the initial mass loading of hematite increased from 0 to 20 mmol of Fe(III)/L, the rate and extent of U(IV) reoxidation increased. Subsequent addition of hematite [15 mmol of Fe(III)/L] to stationary-phase cultures containing microbially reduced U(IV) also resulted in rapid reoxidation to U(VI). Analysis by U L3-edge X-ray absorption near-edge spectroscopy (XANES) of microbially reduced U particles yielded spectra similar to that of natural uraninite. Observations by high-resolution transmission electron microscopy, selected area electron diffraction, and energy-dispersive X-ray spectroscopic analysis confirmed that precipitated U associated with cells was uraninite with particle diameters of 3-5 nm. By the same techniques, iron sulfide precipitates were found to have a variable Fe and S stoichiometry and were not associated with cells.     

Structural identification of isomallotusinin and other phenolics in Phyllanthus emblica L. fruit hull

The air-dried fruit hull of Phyllanthus emblica L. was extracted with 95% ethanol, and then the extract was partitioned by diethyl ether and ethyl acetate (EA). The EA fraction was then subjected to separation and purification using silica gel and Sephadex LH-20 column chromatography repeatedly to obtain five hydrolysable tannins. They were identified as mucic acid 1,4-lactone 3-o-gallate (C1), isocorilagin (C2), chebulanin (C3), chebulagic acid (C4) and isomallotusinin (C5) using mass spectrometry and nuclear magnetic resonance (NMR) spectrometry. Isomallotusinin and chebulanin were identified from emblica dried fruit hull for the first time, and isomallotusinin was the first time identified from Phyllanthus. Furthermore, the antioxidant abilities of these hydrolysable tannins were investigated using DPPH and ABTS⁺ radical scavenging systems. All hydrolysable tannins showed strong DPPH and ABTS⁺ radical scavenging activities. Isomallotusinin and chebulagic acid exhibited the highest antioxidant activity compared to other purified compounds tested.     

Sustainability of uranium mining and milling: toward quantifying resources and eco-efficiency

The mining of uranium has long been a controversial public issue, and a renewed debate has emerged on the potential for nuclear power to help mitigate against climate change. The central thesis of pro-nuclear advocates is the lower carbon intensity of nuclear energy compared to fossil fuels, although there remains very little detailed analysis of the true carbon costs of nuclear energy. In this paper, we compile and analyze a range of data on uranium mining and milling, including uranium resources as well as sustainability metrics such as energy and water consumption and carbon emissions with respect to uranium production-arguably the first time for modern projects. The extent of economically recoverable uranium resources is clearly linked to exploration, technology, and economics but also inextricably to environmental costs such as energy/water/chemicals consumption, greenhouse gas emissions, and social issues. Overall, the data clearly show the sensitivity of sustainability assessments to the ore grade of the uranium deposit being mined and that significant gaps remain in complete sustainability reporting and accounting. This paper is a case study of the energy, water, and carbon costs of uranium mining and milling within the context of the nuclear energy chain.     

Uranium immobilization by sulfate-reducing biofilms

Hexavalent uranium [U(VI)] was immobilized using biofilms of the sulfate-reducing bacterium (SRB) Desulfovibrio desulfuricans G20. The biofilms were grown in flat-plate continuous-flow reactors using lactate as the electron donor and sulfate as the electron acceptor. U(VI)was continuously fed into the reactor for 32 weeks at a concentration of 126 microM. During this time, the soluble U(VI) was removed (between 88 and 96% of feed) from solution and immobilized in the biofilms. The dynamics of U immobilization in the sulfate-reducing biofilms were quantified by estimating: (1) microbial activity in the SRB biofilm, defined as the hydrogen sulfide (H2S) production rate and estimated from the H2S concentration profiles measured using microelectrodes across the biofilms; (2) concentration of dissolved U in the solution; and (3) the mass of U precipitated in the biofilm. Results suggest that U was immobilized in the biofilms as a result of two processes: (1) enzymatically and (2) chemically, by reacting with microbially generated H2S. Visual inspection showed that the dissolved sulfide species reacted with U(VI) to produce a black precipitate. Synchrotron-based U L3-edge X-ray absorption near edge structure (XANES) spectroscopy analysis of U precipitated abiotically by sodium sulfide indicated that U(VI) had been reduced to U(IV). Selected-area electron diffraction pattern and crystallographic analysis of transmission electron microscope lattice-fringe images confirmed the structure of precipitated U as being that of uraninite.     

Chemical and analytical screening of some edible mushrooms

Fractionation of extracts of the edible mushroom, Volvariella volvacea, led to the isolation of two heterocyclic carboxylic acids, namely pyridine-3-carboxylic acid [nicotinic acid, (5)] and pyrazole-3(5)-carboxylic acid (6) and the four steroidal metabolites ergosterol (1), 5-dihydroergosterol (2), ergosterol peroxide (3), cerevisterol (4). Significantly, compound (6) was identified for the first time, to our knowledge, in the mushroom kingdom and is of taxonomic significance. Compounds (2–4) were isolated for the first time from the Volvariella genus. In view of the structural similarity of compound (6) to pyrazole-3-carboxylic acids, which act as agonists for nicotinic acid receptors, the levels of compounds (5) and (6) were estimated for the first time using HPTLC in V. volvacea and two other edible mushrooms, namely Agaricus bisporus and Calocybe indica. Significant levels of compound (5) were found in C. indica, and compound (6) was found in abundance in A. bisporus. Correlations are suggested between the occurrence of these compounds in mushrooms and consumption as well as beneficial health effects of this food.     

VARIETAL AROMA COMPOUNDS OF VITIS VINIFERA CV. KHAMRI GROWN IN TUNISIA

ABSTRACT

The aroma composition of the grape juice of Khamri, a native variety of Vitis vinifera grown in Tunisia, was investigated for the first time. A total of 27 free and 20 glycosidically bound compounds were identified by gas chromatography‐mass spectrometry. According to the obtained results, the aroma compounds were C6 alcohols, benzene compounds, terpenes, acids and norisoprenoids. On the basis of gas chromatography‐olfactometry, these compounds were grouped, according to volatiles exhibiting the identical odor quality, into 10 groups of the same character (aromatic series) as a way of establishing an aroma profile for the studied variety. The high glycosidically bound norisoprenoid concentrations and the absence of the bound form of the acids were a positive factor for the Khamri variety potential aroma.

PRACTICAL APPLICATIONS

This article aimed on the identification of the unknown Tunisian grapevine varieties that are very well adapted to the arid conditions and that could have a good quality. The discovery of unknown autochthonous grapevine varieties with good aroma and pomological characteristics could be of great importance for the agriculture sector all over the world especially the arid regions.

     

Validated measurements of the uranium isotopic signature in human urine samples using magnetic sector-field inductively coupled plasma mass spectrometry

Increased interest in measuring uranium isotope ratios in environmental samples (biological materials, soils, dust particles, water) has come from the necessity to assess the health impact of the use of depleted uranium (DU) based ammunitions during recent military conflicts (e.g., Gulf war, Kosovo) and from the need to identify nondeclared nuclear activities (nuclear safeguards). In this context, very important decisions can arise which have to be based on measurement data of nondisputable uncertainty. The present study describes the certification to 2.5% (k = 2) relative combined uncertainty of n(235U)/n(238U) at ultralow uranium levels (approximately 5-20 pg g(-1)) in human urine samples. After sample decomposition and matrix separation, the isotope ratios were measured by means of a single-detector magnetic sector-field inductively coupled plasma mass spectrometry instrument fitted with an ultrasonic nebulizer. Correction for mass discrimination effects was obtained by means of the certified isotopic reference material IRMM-184. The analytical procedure developed was validated in three complementary ways. First, all major sources of uncertainty were identified and propagated together following the ISO/GUM guidelines. Second, this quality was controlled with a matrix matching NUSIMEP-3 sample (approximately 0.06-0.7% difference from certified). Third, the instrumental part of the procedure was proven to be reproducible from the confirmation of the results obtained for three samples remeasured 7 months later (approximately 1.5% difference). The results obtained for 33 individuals indicated that none seemed to have been exposed to contamination by DU.     

Composition of the essential oils of Sideritis cladestina ssp Cyllenea and Sideritis sipylea

The quantitative composition of the essential oils from two different Sideritis (Labiatae) species (S cladestina spp cyllenea and S sipylea) is reported. Over 80 components have been identified in each species by means of GC and GC? MS analysis, and a number of substances have been identified for the first time in these essential oils.     

Occurrence of sotolon,abhexon and theaspirane‐derived molecules in Gueuze beers. Chemical similarities with ‘yellow wines’

The typical Belgian Gueuze beers are produced with aged hop from a grist of malt and wheat, according to a very long oxidation process (extended boiling, cooling overnight in an open‐air container, oak‐barrel ageing). Two theaspirane‐oxidation‐derived products, dihydrodehydro‐β‐ionone and 4‐hydroxy‐7,8‐dihydro‐β‐ionone, are evidenced here for the first time in Gueuze beers. Both compounds have been recently identified in oxidative wines such as Jura Flor‐Sherry and Sauternes wines. Another analogy with Jura Flor‐Sherry wines was the presence of the nutty/curry odorants sotolon and abhexon, although at lower concentrations. Copyright © 2012 The Institute of Brewing & Distilling     

Acids in chicory roots and malt 1. Identification in roasted products and method of determination

A determination method for non-volatile acids in roasted chicory roots and roasted barley malt is described. The clean-up was accomplished using preparative gel electrophoresis followed by gas chromatography (GC)/mass spectrometry of the methoximes (ketonic acids) or trimethylsilyl derivatives (all acids). Using these methods, 64 non-volatile acids have been identified in chicory (among those 48 for the first time). In barley malt 60 acids could be detected (47 for the first time). Ten commercially unavailable substances have been synthesized. Most of the acids have been quantified using the same clean-up and GC/selected ion monitoring or GC/flame ionisation detection (FID). The recoveries range from 41 to 105% (average 79%), the detection limits from 1 to 72g/kg being somewhat lower for the FID and considerably lower for malt, and the relative standard deviations from 1 to 10% for the main acids, from 2 to 89% for the minor acids and from 8 to 133% for the trace acids. In addition formic and acetic acids were determined by isotachophoresis.

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Säuren in Zichorienwurzeln und Malz: 1. Identifizierung in Röstprodukten und Bestimmungsmethode

Zusammenfassung In Röstzichorie wurden 64 nichtflüchtige Säuren, davon 48 neu in Zichorie, in Röstmalz 60 Säuren, davon 47 neu in Malz, durch GC/MS nach Abtrennung durch präparative Gelelektrophorese, Überführung der Oxosäuren in ihre Methoxime und Trimethylsilylierung identifiziert. 10 im Handel nicht erhältliche Säuren wurden synthetisiert. Die meisten Säuren wurden quantitativ nach derselben Abtrennung mittels GC/SIM oder GC/FID bestimmt. Die Wiederfindungsraten liegen zwischen 41 und 105%, im Mittel bei 79%, die Nachweisgrenzen zwischen 1 und 72 g/kg (etwas niedriger bei GC/FID, erheblich niedriger bei Malz), die Variationskoeffizienten zwischen 1 und 10% bei den Hauptsäuren, zwischen 2 und 89% bei den Minorsäuren und zwischen 8 und 133% bei den Spurensäuren. Zusätzlich wurden Ameisen- und Essigsäure durch Isotachophorese bestimmt.

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Excerpt from doctoral thesis: Säuren in Zichorie und Malz of H. Barlianto [13]