Regulation of phosphorus bioavailability by iron nanoparticles in a monomictic lake.

Dissolved reactive phosphorous (DRP) in lake systems is conventionally considered to predominate over other dissolved P species, however, this view neglects an important set of interactions that occurs between P and reactive iron hydroxide surfaces. This study addresses the coupling of P with dispersed iron nanoparticles in lakes, an interaction that may fundamentally alter the bioavailability of P to phytoplankton. We used diffusive gradients in thin films (DGT) and ultrafiltration to study Fe-P coupling in the water column of a monomictic lake over a hydrological year. Fe and P were predominantly colloidal (particle diameters > ~5 nm < ~20 nm) in both oxic epilimnetic and anaerobic hypolimnetic waters, but they were both DGT-labile under sub-oxic conditions, consistent with diffusion and dissolution of Fe-and-P-bearing colloids within the DGT diffusive gel. During peak stratification, increases in Fe and P bioavailability were spatially and temporally coincident with Fe nanoparticle dissolution and the formation of a deep chlorophyll maximum at 5-8 m depth. These results provide a window into the coupling and decoupling of P with mobile iron colloids, with implications for our understanding of the behaviour of nutrients and their influence on phytoplankton community dynamics.

Effect of phosphate buffer on aggregation kinetics of citrate-coated silver nanoparticles induced by monovalent and divalent electrolytes.

The attachment efficiency (α) is an important parameter that can be used to characterize nanoparticle (NPs) aggregation behavior and has been a topic of discussion of several papers in the past few years. The importance of α is because it is one of the key parameters that can be used to model NP environmental fate and behavior. This study uses UV-vis and laser Doppler electrophoresis to monitor the aggregation behavior of citrate-coated silver nanoparticles (cit-AgNPs) induced by Na+ and Ca2+ as counter ions in the presence and absence of Suwannee River fulvic acid (SRFA) as a surrogate of natural organic matter and different concentrations of phosphate buffer (0-1mM). Results demonstrate that phosphate buffer, which serves to maintain pH nearly constant over the course of a reaction, is an important determinant of NP aggregation behavior. Increasing phosphate buffer concentration results in a decrease in the critical coagulation concentrations (CCC) of cit-AgNPs to lower counter ion concentration and an increase of α at the same counter ion concentration, both in the absence and presence of SRFA. SRFA stabilizes AgNPs and increases the CCC to higher counter ion concentrations. The outcome of this study can be used to rationalize the variation in α and CCC values reported in the literature for NPs with similar physicochemical properties, where different α and CCC values are reported when different types of buffers and buffer concentrations are used in different studies.

The concentration-dependent aggregation of Ag NPs induced by cystine.

Cystine is widely used in cell culture media. Cysteine, the reduced form of cystine, is widely used to scavenge dissolved Ag in eco-toxicological studies to differentiate dissolved vs. nanoparticle uptake and toxicity. However, little is known about the impact of cysteine and cystine on the aggregation behavior of Ag NPs, in particular as a function of Ag NP concentration. Herein, we investigate how cystine (0-300µM) affects the stability of citrate-, polyvinylpyrrolidone-, and polyethylene glycol-coated silver nanoparticles (cit-Ag NPs, PVP-Ag NPs and PEG-Ag NPs, respectively) with and without Suwannee River fulvic acid (SRFA) as a function of Ag NPs concentration using UV-vis spectroscopy at environmentally and ecotoxicologically relevant Ag NP concentrations (ca. 125-1000µgL(-1)). The results demonstrate, for the first time, the concentration-dependent aggregation of cit-Ag NPs in the presence of cystine with a shift in the critical coagulation concentration (CCC) to lower cystine concentrations at lower cit-Ag NP concentrations. At the highest cit-Ag NP concentration (1000µgL(-1)), reaction limited aggregation was only observed and no CCC was measured. SRFA slowed the aggregation of cit-Ag NPs by cystine and aggregation occurred in reaction limited aggregation (RLA) regime only. No CCC value was measured in the presence of SRFA. Cystine replaces citrate, PVP and PEG coatings, resulting in aggregation of both electrostatically and sterically stabilized Ag NPs. These findings are important in understanding the factors determining the behavior of Ag NPs in cell culture media. Also due to the similarity between cystine and cysteine, these results are important in understanding the uptake and toxicity of Ag NPs vs. Ag ions, and suggest that the reduction of the toxicity of Ag NPs in the presence of cysteine could be due to a combined effect of scavenging Ag(+) ions and Ag NP aggregation in the presence of cysteine.

An electron microscopy based method for the detection and quantification of nanomaterial number concentration in environmentally relevant media.

Improved detection and characterization of nanomaterials (NMs) in complex environmental media requires the development of novel sampling approaches to improve the detection limit to be close to environmentally realistic concentrations. Transmission electron microscopy (TEM) is an indispensable metrological tool in nanotechnology and environmental nanoscience due to its high spatial resolution and analytical capabilities when coupled to spectroscopic techniques. However, these capabilities are hampered by the conventional sample preparation methods, which suffer from low NM recovery. The current work presents a validated, fully quantitative sampling technique for TEM that overcomes conventional sample preparation shortcomings, and thus enables the use of TEM for measurement of particle number concentration and their detection in complex media at environmentally realistic concentrations. This sampling method is based on ultracentrifugation of NMs from suspension onto a poly-l-lysine (PLL) functionalized TEM grid, using active deposition (by ultracentrifugation) and retention (by PLL interactions with NM surface) of NMs on the substrate, enabling fully quantitative analysis. Similar analysis with AFM was satisfactory in simple media but the lack of chemical-selectivity of AFM limits its applicability for the detection of NMs in complex environmental samples. The sampling approach was validated using both citrate- and PVP-coated AuNMs in pure water, which demonstrated an even distribution of NM on the TEM grid and high NM recovery (80-100%) at environmentally relevant NM concentrations (ca. 0.20-100 µg L(-1)). The applicability of the sampling method to complex environmental samples was demonstrated by the quantification of particle number concentration of AuNMs in EPA soft water (with and without Suwannee River fulvic acid) and lake water. This sample preparation approach is also applicable to other types of NMs with some modifications (e.g. centrifugation force and time) to insure full sample recovery. This TEM sampling method is key to the accurate quantification of NM number concentration, and therefore to improving our understanding of environmental fate, behavior, effects and dose of NMs.

Evaluation of charge and agglomeration behavior of TiO2 nanoparticles in ecotoxicological media.

The dynamic nature of nanoparticle (NP) agglomeration behavior is of paramount interest to many current studies in environmental nanoscience and nano(eco)toxicology because agglomeration affects the NP bioavailability and toxicity. The present study investigates the surface charge and agglomeration behavior of TiO2 NPs in four different ecotoxicological media (OECD algae, OECD L_variegatus, hardwater and plant media) and two different electrolytes KCl (200 mM) and CaCl2 (50 mM). TiO2 NPs were positively charged, and the zeta potential varied from +1.9 mV in hardwater (at pH7.1) to +24.5 mV in CaCl2 electrolyte (at pH7.4) in all media except algae media, where the zeta potential was -6.7 mV (at pH7.7). Despite the differences in the pH and the surface charge of TiO2 NPs in the different media, an immediate agglomeration of the NPs in all standard ecotoxicological media was observed with aggregate sizes in the micrometer scale, as the measured zeta potentials were insufficient to prevent TiO2 NP agglomeration. The isoelectric point (pHiep) of TiO2 NPs in the studied media varied in the range (6.8-7.6), which was attributed to preferential association of anions and cations to TiO2; that is the pHiep decreases with the increased concentration of Cl and increases with the increased concentrations of Na and Mg. Despite the complexity of the ecotoxicological media and the presence of a mixture of different monovalent and divalent electrolytes, the agglomeration kinetics in the media follows the DVLO theory where two distinct agglomeration rates (slow, reaction limited regime and fast, diffusion limited regime) were observable. The critical coagulation concentration (CCC) of TiO2 NPs in the ecotoxicological media varied from 17.6 to 54.0% v/v standard media in UHPW, due to differences in media pH and TiO2 NP surface charge. In the ecotoxicological media (hardwater, L-variegatus and plant), where TiO2 NPs are positively charged, the CCC decrease with the increased divalent anions (act as counter ions) concentration in the media, again in good agreement with the DLVO theory.

Transformations of citrate and Tween coated silver nanoparticles reacted with Na2S.

Silver nanoparticles (Ag NPs) are susceptible to transformations in environmental and biological media such as aggregation, oxidation, dissolution, chlorination, sulfidation, formation/replacement of surface coatings following interaction with natural organic matter (NOM). This paper investigates the impact of surface coating and Suwannee River fulvic acid (SRFA) on the transformations and behavior of Ag NPs (citrate coated and Tween coated; cit-Ag NPs and Tween-Ag NPs, respectively), following reaction with different concentrations of Na2S solution (as a source of sulfide species, H2S and HS(-)). These transformations and the dominant mechanisms of transformations were investigated using UV-vis and scanning transmission electron microscopy coupled with electron energy loss spectroscopy. Here, we have shown that Ag NP surface coating impacts their dissolution following dilution in ultrahigh purity water, with higher extent of dissolution of Tween-Ag NPs compared with cit-Ag NPs. Tween-Ag NPs are susceptible to dissolution following their sulfidation at low S/Ag molar ratio. Suwannee River fulvic acid (SRFA) slows down the dissolution of Tween-Ag NPs at low sulfide concentrations and reduces the aggregation of cit-Ag NP in the presence of sodium sulfide. Sulfidation appears to occur by direct interaction of sulfide species with Ag NPs rather than by indirect reaction of sulfide with dissolved Ag species subsequent to dissolution. Furthermore, the sulfidation process results in the formation of partially sulfidized Ag NPs containing unreacted (metallic) subgrains at the edge of the NPs for Tween-Ag NPs in the presence of high sulfide concentration (2000nM Na2S), which occurred to less extent at lower Na2S concentration for Tween-Ag NPs and at all concentrations of Na2S for cit-Ag NPs. Thus, sulfidized Ag NPs may preserve some of the properties of the Ag NPs such as their potential to shed Ag(+) ions and their toxic potential of Ag NPs.

Quantitative measurement of the nanoparticle size and number concentration from liquid suspensions by atomic force microscopy.

Microscopy techniques are indispensable to the nanoanalytical toolbox and can provide accurate information on the number size distribution and number concentration of nanoparticles (NPs) at low concentrations (ca. ppt to ppb range) and small sizes (ca. <20 nm). However, the high capabilities of microscopy techniques are limited by the traditional sample preparation based on drying a small volume of suspension of NPs on a microscopy substrate. This method is limited by low recovery of NPs (ca. <10%), formation of aggregates during the drying process, and thus, the complete misrepresentation of the NP suspensions under consideration. This paper presents a validated quantitative sampling technique for atomic force microscopy (AFM) that overcomes the above-mentioned shortcomings and allows full recovery and representativeness of the NPs under consideration by forcing the NPs into the substrate via ultracentrifugation and strongly attaches the NPs to the substrate by surface functionalization of the substrate or by adding cations to the NP suspension. The high efficiency of the analysis is demonstrated by the uniformity of the NP distribution on the substrate (that is low variability between the number of NPs counted on different images on different areas of the substrate), the high recovery of the NPs up to 71%) and the good correlation (R > 0.95) between the mass and number concentrations. Therefore, for the first time, we developed a validated quantitative sampling technique that enables the use of the full capabilities of microscopy tools to quantitatively and accurately determine the number size distribution and number concentration of NPs at environmentally relevant low concentrations (i.e. 0.34-100 ppb). This approach is of high environmental relevance and can be applied widely in environmental nanoscience and nanotoxicology for (i) measuring the number concentration dose in nanotoxicological studies and (ii) accurately measuring the number size distribution of NPs; both are key requirements for the implementation of the European Commission recommendation for definition of nanomaterials.

Effect of monovalent and divalent cations, anions and fulvic acid on aggregation of citrate-coated silver nanoparticles.

The dynamic nature of nanoparticle (NP) aggregation behavior is of paramount interest to many current studies in environmental and toxicological nanoscience. The present study seeks to elucidate the influence that different electrolytes have on the aggregation of citrate-coated silver NPs (cit-AgNPs). The use of both UV-vis spectroscopy and dynamic light scattering (DLS, both z-average hydrodynamic diameter (z-dh) and size distribution analysis data) allowed improvement in the data quality and interpretation as compared to other studies using only DLS and reporting solely the z-dh, as the change in the z-dh can be related to analytical errors and uncertainties rather than only aggregation or dissolution of NPs. Divalent cations (CaCl2, Ca(NO3)2, CaSO4, MgCl2 and MgSO4) have stronger influence (ca. 50-65 fold) on aggregation of cit-AgNPs as compared to monovalent cations (NaCl, NaNO3, Na2SO4), as expected. For electrolytes with monovalent cations, there was no specific ion effect of nitrate and sulfate anions. However, the addition of chloride anions resulted in enhanced apparent aggregation, possibly due to the formation of AgCl NPs that sorb/attach to the surface of cit-AgNPs. Suwannee River fulvic acid enhances the stability of cit-AgNPs and shifts the critical coagulation concentrations to higher electrolyte concentrations for all types of electrolytes. Aggregation kinetics in the presence of mixture of monovalent and divalent cations is additive and controlled by the dominant cations. An empirical formula (αmixture=αNa+(50 to 65)Ca) is proposed that reproduces the effect of mixtures of electrolytes in the presence of humic substances and cations that can be used to help predict the aggregation behavior of cit-AgNPs in environmental and ecotoxicological media.

Rationalizing nanomaterial sizes measured by atomic force microscopy, flow field-flow fractionation, and dynamic light scattering: sample preparation, polydispersity, and particle structure.

This study aims to rationalize the variability in the measured size of nanomaterials (NMs) by some of the most commonly applied techniques in the field of nano(eco)toxicology and environmental sciences, including atomic force microscopy (AFM), dynamic light scattering (DLS), and flow field-flow fractionation (FlFFF). A validated sample preparation procedure for size evaluation by AFM is presented, along with a quantitative explanation of the variability of measured sizes by FlFFF, AFM, and DLS. The ratio of the z-average hydrodynamic diameter (d(DLS)) by DLS and the particle height by AFM (d(AFM)) approaches 1.0 for monodisperse samples and increases with sample polydispersity. A polydispersity index of 0.1 is suggested as a suitable limit above which DLS data can no longer be interpreted accurately. Conversion of the volume particle size distribution (PSD) by FlFFF-UV to the number PSD reduces the differences observed between the sizes measured by FlFFF (d(FlFFF)) and AFM. The remaining differences in the measured sizes can be attributed to particle structure (sphericity and permeability). The ratio d(FlFFF)/d(AFM) approaches 1 for small ion-coated NMs, which can be described as hard spheres, whereas d(FlFFF)/d(AFM) deviates from 1 for polymer-coated NMs, indicating that these particles are permeable, nonspherical, or both. These findings improve our understanding of the rather scattered data on NM size measurements reported in the environmental and nano(eco)toxicology literature and provide a tool for comparison of the measured sizes by different techniques.

Flow field-flow fractionation for the analysis and characterization of natural colloids and manufactured nanoparticles in environmental systems: a critical review.

The use of flow field flow fractionation (FlFFF) for the separation and characterization of natural colloids and nanoparticles has increased in the last few decades. More recently, it has become a popular method for the characterization of manufactured nanoparticles. Unlike conventional filtration methods, FlFFF provides a continuous and high-resolution separation of nanoparticles as a function of their diffusion coefficient, hence the interest for use in determining particle size distribution. Moreover, when coupled to other detectors such as inductively coupled plasma-mass spectroscopy, light scattering, UV-absorbance, fluorescence, transmission electron microscopy, and atomic force microscopy, FlFFF provides a wealth of information on particle properties including, size, shape, structural parameters, chemical composition and particle-contaminant association. This paper will critically review the application of FlFFF for the characterization of natural colloids and natural and manufactured nanoparticles. Emphasis will be given to the detection systems that can be used to characterize the nanoparticles eluted from the FlFFF system, the obtained information and advantages and limitation of FlFFF compared to other fractionation and particle sizing techniques. This review will help users understand (i) the theoretical principles and experimental consideration of the FlFFF, (ii) the range of analytical tools that can be used to further characterize the nanoparticles after fractionation by FlFFF, (iii) how FlFFF results are compared to other analytical techniques and (iv) the range of applications of FlFFF for natural and manufactured NPs.

Algal testing of titanium dioxide nanoparticles--testing considerations, inhibitory effects and modification of cadmium bioavailability.

The ecotoxicity of three different sizes of titanium dioxide (TiO(2)) particles (primary particles sizes: 10, 30, and 300nm) to the freshwater green alga Pseudokirchneriella subcapitata was investigated in this study. Algal growth inhibition was found for all three particle types, but the physiological mode of action is not yet clear. It was possible to establish a concentration/dose-response relationship for the three particle sizes. Reproducibility, however, was affected by concentration-dependent aggregation of the nanoparticles, subsequent sedimentation, and possible attachment to vessel surfaces. It is also believed that heteroaggregation, driven by algal exopolymeric exudates, is occurring and could influence the concentration-response relationship. The ecotoxicity of cadmium to algae was investigated both in the presence and absence of 2mg/L TiO(2). The presence of TiO(2) in algal tests reduced the observed toxicity due to decreased bioavailability of cadmium resulting from sorption/complexation of Cd(2+) ions to the TiO(2) surface. However, for the 30nm TiO(2) nanoparticles, the observed growth inhibition was greater than what could be explained by the concentration of dissolved Cd(II) species, indicating a possible carrier effect, or combined toxic effect of TiO(2) nanoparticles and cadmium. These results emphasize the importance of systematic studies of nanoecotoxicological effects of different sizes of nanoparticles and underline the fact that, in addition to particle toxicity, potential interactions with existing environmental contaminants are also of crucial importance in assessing the potential environmental risks of nanoparticles.

Nanoparticles: structure, properties, preparation and behaviour in environmental media.

There is increasing interest and need to develop a deeper understanding of the nature, fate and behaviour of nanoparticles in the environment. This is driven by the increased use of engineered nanoparticles and the increased pressure to commercialise this growing technology. In this review we discuss the key properties of nanoparticles and their preparation and then discuss how these factors can play a role in determining their fate and behaviour in the natural environment. Key focus of the discussion will relate to the surface chemistry of the nanoparticle, which may interact with a range of molecules naturally present in surface waters and sediments. Understanding these factors is a core goal required for understanding the final fate of nanomaterials and predicting which organisms are likely to be exposed to these materials.

Size fractionation and characterization of natural aquatic colloids and nanoparticles.

Atomic force microscopy (AFM) was used to image and quantify natural nanoparticles (prefiltered <25 nm) from three different freshwater sites (Vale Lake, Bailey Brook and Tern Rivers). Four fractions were analysed by AFM; the prefiltered fraction (<25 nm) and three fractions collected after separation of this prefiltered sample by flow field-flow fractionation (FlFFF) which corresponds to material which has size ranges of <4.2 nm, 4.2-15.8 nm and 15.8-32.4 nm, as determined by FlFFF theory. The large majority of materials in all samples appeared as <3 nm nanoparticles, nearly spherical and rich in chromophores active at 254 nm UV, which thus correspond to natural organic matter. However, nanoparticles were also imaged up to slightly more than 25 nm in size, indicating a slight disagreement in sizing between filtration and FlFFF. In addition, some particles in certain fractions were found to be covered with a thin film of less than 0.5-1.0 nm. Substantial differences between sites were observed.

Characterization of natural aquatic colloids (<5 nm) by flow-field flow fractionation and atomic force microscopy.

Flow-field flow fractionation (FIFFF) coupled to a UV detector and to atomic force microscopy (AFM) has been used, for the first time, to characterize ultrafine natural colloids (<5 nm) from selected freshwaters. FIFFF-UV measures a "weight diffusion coefficient distribution" and the corresponding "weight hydrodynamic diameter distribution" was calculated by applying FIFFF theory and the Stokes-Einstein equation. In addition, FIFFF has been used to prepare fractions of very narrow size range for AFM analysis. AFM measures number distribution of particle height (related to radius), and these were calculated. Both raw and transformed data show good agreement between the techniques, with conversion of the UV data to a number-weighted distribution giving better agreement and reduced errors. The small differences between the corrected data from UV analysis and the raw AFM data are either due to the fundamental differences in the analytical techniques, that is, measurement of hydrodynamic properties (FIFFF) or properties after sorption to a solid phase (AFM), or are due to the assumptions of the Stokes-Einstein equation not being met, that is, the fine natural colloids are not spherical or permeable. The methodology offers a means of quantifying fine colloid nonsphericity and permeability.

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.

Size fractionation and characterization of natural colloids by flow-field flow fractionation coupled to multi-angle laser light scattering.

Flow-field flow fractionation (FlFFF) coupled to multi-angle laser light scattering (MALLS) was evaluated for size and shape determination of standard spherical and arbitrarily shaped natural colloids. Different fitting methods for light scattering data retrieved from MALLS were evaluated to determine the particle size of spherical standards and natural colloids. In addition, FlFFF was optimized for best fractionation in connection to MALLS, minimal colloids-membrane interaction, and minimal sample losses. FlFFF, calibrated with standard particles, was used to determine hydrodynamic diameter, or radius (D(h) or R(h)), of the fractionated colloids, whereas the MALLS was used to determine root mean square radius of gyration (R(g)) for fractionated colloids. Combining both results, by calculating the R(g)/R(h) ratio, allows an estimation of colloid deviation from the shape of homogeneous sphere. Accordingly, this study demonstrates that, FlFFF-MALLS is a valuable technique for characterizing heterogeneous and arbitrarily shaped natural colloidal particles in terms of size and shape. To check the usefulness of FlFFF-MALLS in natural colloid studies, the technique was used to investigate the sedimentation behavior of extracted soil colloidal particles. Results illustrate that, in a silty till sample, carbonates function as cement between the colloidal particles, and consequently, change their sedimentation behavior. On the other hand, carbonate dissolution generates a more homogeneous colloidal sample.

Natural sample fractionation by FlFFF-MALLS-TEM: sample stabilization, preparation, pre-concentration and fractionation.

Two flow field flow fractionation (FlFFF) systems: symmetrical (SFlFFF) and asymmetrical (ASFlFFF) were evaluated to fractionate river colloids. Samples stability during storage and colloids concentration are the main challenges limiting their fractionation and characterization by FlFFF. A pre-fractionation (<0.45 microm) and addition of a bactericide such as NaN3 into river colloidal samples allowed obtaining stable samples without inducing any modification to their size. Stirred cell ultra-filtration allowed colloidal concentration enrichment of 25-folds. Scanning electron microscope (SEM) micrographs confirmed the gentle pre-concentration of river samples using the ultra-filtration stirred cell. Additionally, larger sample injection volume in the case of SFlFFF and on channel concentration in the case of ASFlFFF were applied to minimize the required pre-concentration. Multi angle laser light scattering (MALLS), and transmission electron microscope (TEM) techniques are used to evaluate FlFFF fractionation behavior and the possible artifacts during fractionation process. This study demonstrates that, FlFFF-MALLS-TEM coupling is a valuable method to fractionate and characterize colloids. Results prove an ideal fractionation behavior in case of Brugeilles sample and steric effect influencing the elution mode in case of Cézerat and Chatillon. Furthermore, comparison of SFlFFF and ASFlFFF fractograms for the same sample shows small differences in particle size distributions.