Sedimentation Field-flow Fractionation Research Articles

Kinetics of chemically mediated neurodegeneration/neuroregeneration of mouse olfactory epithelium: monitoring by hyperlayer sedimentation field flow fractionation

The increase in the incidence of neurodegenerative diseases linked to aging or injury needs to be addressed in research into neuroprotective or neuroregenerative therapies, and requires the development of specific biological models. To achieve this goal we propose (1) the use of the mouse olfactory epithelium as a biological support which specifically exhibits a regenerative or a self-renewing capacity and during the lifetime necessitates the presence of neural stem cells, and (2) the use of an intraperitoneal injection of 2,6-dichlorobenzonitrile (diclobenil) as a chemical inducer of neurodegeneration in olfactory epithelium by selectively killing mature cells. We developed a biological model to follow the processes of neurodegeneration (chemically induced) and neuroregeneration (self-renewal of olfactory epithelium). The purpose of this study was to develop a method to monitor quickly neurodegeneration/neuroregeneration processes in order to further screen protective and regenerative therapies. For this purpose, we used the sedimentation field flow fractionation elution of olfactory epithelium. We obtained specific elution profiles and retention parameters allowing the monitoring of the induction and kinetics of biological processes. The use of insulin-like growth factor1α as a neuroprotective agent in an innovative nebulization protocol showed sedimentation field flow fractionation to be a simple, fast and low-cost method to monitor such a biological event on the scale of an entire organism.

Development and characterization of PLGA nanoparticles as delivery systems of a prodrug of zidovudine obtained by its conjugation with ursodeoxycholic acid

Context: Zidovudine (AZT) is employed against AIDS and hepatitis; its use is limited by active efflux transporters (AETs) that induce multidrug resistance for intracellular therapies and hamper AZT to reach the brain. Ursodeoxycholic acid (UDCA) conjugation with AZT (prodrug UDCA–AZT) allows to elude the AET systems.Objective: To investigate the effect of the Pluronic F68 coating on the loading, release and stability of poly(D,L lactide-co-glicolide) nanoparticles (NPs) embedded with UDCA–AZT.Materials and methods: The mean diameter of the NP prepared by nanoprecipitation or emulsion/solvent evaporation methods was determined using both photon correlation spectroscopy and sedimentation field–flow fractionation; particle morphology was detected by scanning electron microscope. The stability of the free and encapsulated UDCA–AZT was evaluated in rat liver homogenates by high-performance liquid chromatography analysis.Results and discussion: The mean diameter of the NPs was found to be ∼600 nm with a relatively high polydispersity. The NPs obtained by emulsion/solvent evaporation were not able to control the prodrug release, differently from NPs obtained by nanoprecipitation. The presence of the Pluronic coating did not substantially modify the kinetics of the drug release, or the extent of the burst effect that were instead only influenced by the preparation parameters. UDCA–AZT incorporated in the NPs was more stable in the rat liver homogenates than the free prodrug and no influence of the Pluronic coating was observed.Conclusions: Considering the different potential applications of nanoparticles coated and uncoated with Pluronic (brain and macrophage targeting, respectively), both of these nanoparticle systems could be useful in the therapies against HIV.

Determination of size distribution of colloidal TiO2 nanoparticles using sedimentation field-flow fractionation combined with single particle mode of inductively coupled plasma-mass spectrometry

Sedimentation field-flow fractionation (SdFFF) provides mass-based separation of colloidal particles, and thus a size-based separation if the particles are of uniform density. The particle size can be measured from the retention time in SdFFF if the density of the particle is known. In the single-particle mode of inductively coupled plasma-mass spectrometer (SP-ICPMS), the signal intensity depends on the particle size, and the signal frequency is proportional to the particle concentration. In this study, SdFFF was employed for size-based separation and size analysis of two types of TiO2 nanoparticles, rutile and anatase, having rather broad size distributions. SdFFF provided high enough resolution to show the difference in size distributions between two samples, for which dynamic light scattering (DLS) could not show clearly. SdFFF fractions were collected and analyzed by SP-ICPMS to determine size distributions of the SdFFF fractions. The sizes and Ti concentrations of SdFFF fractions of TiO2 colloids determined by SP-ICPMS were well correlated with SdFFF results. Results revealed that SP-ICP could become a useful tool for size-based separation and also for size analysis of various types of colloidal nanoparticles, when it is combined with a particle separator such as SdFFF, especially for samples having relatively broad size distributions.

Kinetic study of aggregation of milk protein and/or surfactant-stabilized oil-in-water emulsions by Sedimentation Field-Flow Fractionation

Milk proteins are able to facilitate the formation and stabilization of oil droplets in food emulsions. This study employed Sedimentation Field-Flow Fractionation (SdFFF) to monitor changes in particle size distribution of freshly prepared emulsions with varying weight contributions of sodium caseinate (SC) and whey protein concentrate (WPC). The effect of the addition of Tween 80 (T) on the initial droplet size was also investigated. The results indicated that emulsifying ability follows the order Tween 80>WPC>SC, with corresponding weight average droplet diameter of 0.319, 0.487 and 0.531μm respectively, when each of the above emulsifiers was used solely. The stability of sodium caseinate emulsions was studied at 30.5 and 80.0°C by measuring the particle size distribution for a period of 70h. Emulsions withstood the temperatures and exhibited an initial increase in particle size distribution caused by heat-induced droplet aggregation, followed by a decrease to approximately the initial droplet size. The rate of droplet aggregation depends on the severity of thermal processing, as revealed by the kinetics of particle aggregation during aging at different temperatures. Comparison of the experimental rate constants found from SdFFF, with those determined theoretically gives invaluable information about the oil droplet stability and the aggregation mechanism. Based on the proposed mechanistic scheme various physicochemical quantities, which are very important in explaining the stability of oil-in-water emulsions, were determined. Finally, the advantages of SdFFF in studying the aggregation of the oil-in-water droplets, in comparison with other methods used for the same purpose, are discussed.

Size characterization by Sedimentation Field Flow Fractionation of silica particles used as food additives

Four types of SiO2, available on the market as additives in food and personal care products, were size characterized using Sedimentation Field Flow Fractionation (SdFFF), SEM, TEM and Photon Correlation Spectroscopy (PCS). The synergic use of the different analytical techniques made it possible, for some samples, to confirm the presence of primary nanoparticles (10nm) organized in clusters or aggregates of different dimension and, for others, to discover that the available information is incomplete, particularly that regarding the presence of small particles. A protocol to extract the silica particles from a simple food matrix was set up, enriching (0.25%, ww−1) a nearly silica-free instant barley coffee powder with a known SiO2 sample. The SdFFF technique, in conjunction with SEM observations, made it possible to identify the added SiO2 particles and verify the new particle size distribution. The SiO2 content of different powdered foodstuffs was determined by graphite furnace atomic absorption spectroscopy (GFAAS); the concentrations ranged between 0.006 and 0.35% (ww−1). The protocol to isolate the silica particles was so applied to the most SiO2-rich commercial products and the derived suspensions were separated by SdFFF; SEM and TEM observations supported the size analyses while GFAAS determinations on collected fractions permitted element identification.

Improved sedimentation field-flow fractionation separation channel for concentrated cellular elution

SdFFF is now commonly used for cell sorting. Nevertheless, as with many other separation methods, SdFFF Hyperlayer elution leads (1) to sample dilution resulting in cell loss which could restrict further use; and (2) to a high output flow rate impacting detector sensitivity and selectivity. In order to limit these problems, we proposed modifications of the SdFFF separation channel consisting both in downscaling and the insertion of an outlet stream splitter. This last system corresponded to a strip which divides the flow rate output into two parts, one containing concentrated cells in a reduced volume and flow rate, the other containing the excess mobile phase useless for further cell manipulation, detection and characterization. For the first time we have shown that splitter implementation and downscaling respected channel flowing and resulted in Hyperlayer elution of around 95% of cells in less than 50% of input flow rate. Improved cell sorting was demonstrated by enrichment (∼10 times) of cancer stem cells from WiDr cells with two times less quantity of injected cells.

Fast astrocyte isolation by sedimentation field flow fractionation

Astrocytes play a key role during central nervous system (CNS) repair and glial scar formation. After CNS damage, an extensive deposition of the extracellular matrix produced by the activated astrocytes limits the extension of the lesion but impairs axon outgrowth and functional recovery. Until now, methods to obtain astrocytes need long culture period and laborious cell culture conditions and do not allow the isolation of pure astrocyte preparation. In this study, we used sedimentation field flow fractionation (SdFFF) to rapidly sort well preserved astrocyte population. Four main cell fractions, the total eluted population (TP), and fractions F1, F2, and F3, were isolated by SdFFF from rat newborn cortex. After elution, cells were cultured for one week, and analyzed by immunocytofluorescence using antibodies against specific epitopes: glial fibrillary acidic protein (GFAP), O4, β-III tubulin, and CD 68, labelling respectively astrocytes, oligodendrocytes, neurons, and microglial cells. SdFFF eluted cells were compared with the cells obtained with the classical method. Results showed that SdFFF appeared to be a rapid (one week) and effective method to sort enriched populations of viable and functional astrocytes. In particular, F1 and F3 fractions contained high percentage of GFAP expressing cells (95.6% and 98.0%, respectively). Results also showed that F1 derived cell cultures contained large astrocytes that spread in the culture dish while in fraction F3 derived cell cultures, astrocytes were small, showing a tendency to aggregate and displaying higher migratory capacities than those of fraction F1. Thanks to SdFFF, isolation of almost pure astrocyte populations was rapidly obtained. In addition, the isolation of different astrocyte subpopulations showing different behaviors offers a new perspective to better understand the glial scar formation and remodeling after CNS damage.

SEDIMENTATION FIELD-FLOW FRACTIONATION AS A TOOL FOR THE STUDY OF MILK PROTEIN-STABILIZED MODEL OIL-IN-WATER EMULSIONS: EFFECT OF PROTEIN CONCENTRATION AND HOMOGENIZATION PRESSURE

Milk protein-stabilized model emulsions were formed using high-pressure homogenization and the effect of protein concentration and homogenization pressure during emulsification on the particle size was studied. Various techniques are available for determining particle size distribution, each one of which has its own advantages and disadvantages. In this study, sedimentation field-flow fractionation was employed for the size characterization of oil-in water emulsion droplets. Increasing protein content results in significant reduction in emulsion particle size for the concentration range (0.5–3.0% w/w) employed in this study. Low protein content (<1%) may be correlated with bridging flocculation leading to increased particle size, as indicated by optical microscopy. Similarly, increasing pressure during the homogenization process results in decreasing significantly the particle size of the oil-in-water emulsions, for the pressure range (20–60 MPa) utilized in this study. Increased heating associated with high levels of pressure during the homogenization process, can result in changes in the oil or protein structure, which in turn may have an impact on the physicochemical properties of the oil-in-water emulsions on a long-term basis. The results of this study indicate that sedimentation field-flow fractionation can be employed to reliably monitor changes in particle size of model emulsions induced by protein concentration and homogenization pressure. The technique can be a valuable tool for understanding the properties of colloidal systems and therefore its implementation in emulsion/dispersion technology laboratories is certainly justified.

Steric 모드의 침강장-흐름 분획법을 이용한 황사의 특성분석

황사입자들은 수 나노미터에서 수 마이크론 사이의 크기를 가지는 것으로 알려져 있다. 황사가 환경 및 인체 건강에 미치는 영향은 황사 입자의 크기에 의존한다. 입자가 작을수록 멀리까지 이동하며, 인체의 호흡기관 깊숙이 침투한다. 침강장-흐름 분획법(sedimentation field-flow fractionation, SdFFF)은 채널 내 포물선형태의 흐름(parabolic flow profile)과 외부에서 가해지는 원심력의 상호작용을 이용하여 나노 및 마이크론 크기의 입자들의 분리를 제공한다. 본 연구에서는 황사입자의 크기별 분리와 특성분석을 위한 steric 모드 침강장-흐름 분획법(Sd/StFFF)의 응용 가능성을 테스트하였다. 이를 위하여 다양한 Sd/StFFF 파라미터들을(유속, stop-flow time, 원심력의 세기, 등) 최적화 하였다. Sd/StFFF 보정곡선의 <TEX>$R^2$</TEX>값은 0.9983으로 높은 직선성을 보였으며, 실험결과는 Sd/StFFF가 마이크론 입자의 크기별 분리에 우수함을 보여주었다. 광학현미경(optical microscopy, OM)을 이용하여 황사입자들의 크기와 모양을 조사하였다. 황사가 진할 때에는 약할 때보다 입자크기가 증가함을 보여주었다. 또한 비가 올 때에는 건조할 때보다 입자크기가 감소하였는데, 이는 입자 표면에 흡착되어 있는 성분들이 빗물에 의해 제거되었기 때문인 것으로 보여진다. 본 연구의 결과는 Sd/StFFF가 황사와 같이 환경입자의 크기특성분석에 유용함을 보여준다. Asian dust particles are known to have sizes ranging from a few nanometers up to about a few micrometers. The environmental and health effects depend on the size of the dust particles. The smaller, the farther they are transported, and the deeper they penetrate into the human respiratory system. Sedimentation field-flow fractionation (SdFFF) provides separation of nano to microparticles using a combination of centrifugal force and parabolic laminar flow in a channel. In this study, the steric mode of SdFFF (Sd/StFFF) was tested for size-based separation and characterization of Asian dust particles. Various SdFFF experimental parameters including flow rate, stop-flow time and field strength of the centrifugal field were optimized for the size analysis of Asian dust. The Sd/StFFF calibration curve showed a good linearity with <TEX>$R^2$</TEX> value of 0.9983, and results showed an excellent capability of Sd/StFFF for a size-based separation of micron-sized particles.The optical microscopy (OM) was also used to study the size and the shape of the dust particles. The size distributions of the samples collected during a thick dust period were shifted towards larger sizes than those of the samples collected during thin dust periods. It was also observed that size distribution of the sample collected during dry period shifts further towards larger sizes than that of the samples collected during raining period, suggesting the sizes of the dust particle decrease during raining periods as the components adsorbed on the surface of the dust particles were removed by the rain water. Results show Sd/StFFFis a useful tool for size characterization of environmental particles such as the Asian dust.

Autophagic Subpopulation Sorting by Sedimentation Field-Flow Fractionation

The development of hypoxic areas often takes place in solid tumors and leads cells to undergo adaptive signalization like autophagy. This process is responsible for misfolded or aggregated proteins and nonfunctional organelle recycling, allowing cells to maintain their energetic status. However, it could constitute a double-edged pathway leading to both survival and cell death. So, in response to stress such as hypoxia, autophagic and apoptotic cells are often mixed. To specifically study and characterize autophagic cells and the process, we needed to develop a method able to (1) isolate autophagic subpopulation and (2) respect apoptotic and autophagic status. Sedimentation field-flow fractionation (SdFFF) was first used to monitor physical parameter changes due to the hypoxia mimetic CoCl(2) in the p53 mutated SKNBE2(c) human neuroblastoma cell line. Second, we showed that "hyperlayer" elution is able to prepare autophagic enriched populations, fraction (F3), overexpressing autophagic markers (i.e., LC3-II accumulation and punctiform organization of autophagosomes as well as cathepsin B overactivity). Conversely, the first eluted fraction exhibited apoptotic markers (caspase-3 activity and Bax increased expression). For the first time, SdFFF was employed as an analytical tool in order to discriminate apoptotic and autophagic cells, thus providing an enriched autophagic fraction consecutively to a hypoxic stress.

Clotrimazole nanoparticle gel for mucosal administration

In this study a formulation suitable to be applied on oral and/or vaginal mucosa has been developed for the treatment of fungal infections.The aim of the research is a comparison between clotrimazole (CLO) containing semisolid formulations based on monoolein aqueous dispersion (MAD) or nanostructured lipid carrier (NLC). MAD and NLC have been characterized in terms of morphology and dimensional distribution by cryogenic Transmission Electron Microscopy (cryo-TEM) and Photon Correlation Spectroscopy (PCS). CLO was encapsulated with high entrapment efficiency both in MAD and in NLC, according to Sedimentation Field Flow Fractionation (SdFFF) combined with HPLC. CLO recovery in MAD and NLC has been investigated by time.In order to obtain formulations with suitable viscosity for mucosal application, MAD was diluted with a carbomer gel, while NLC was directly viscosized by the addition of poloxamer 407 in the dispersion. The rheological properties of MAD and NLC after viscosizing have been investigated.Franz cell has been employed to study CLO diffusion from the different vehicles, evidencing diffusion rates from MAD and NLC superimposable to that obtained using Canesten®.An anticandidal activity study demonstrated that both CLO-MAD and CLO-NLC were more active against Candida albicans with respect to the pure drug.

Effect of Surfactant on Retention Behaviors of Polystyrene Latex Particles in Sedimentation Field-Flow Fractionation: Effective Boundary Slip Model Approach

A retention theory in sedimentation field-flow fractionation (SdFFF) was developed by exploiting the effective slip boundary condition (BC) that allows a finite velocity for particles to have at the wall, thereby alleviating the limitations set by the no-slip BC constraint bound to the standard retention theory (SRT). This led to an expression for the retention ratio R as R = (R(o) + v*(b))/(R(o) + v*(b)), where R(o) is the sterically corrected SRT retention ratio and v*(b) is the reduced boundary velocity. Then, v*(b) was modeled as v*(b) = v*(b,o)/[1 + (7K*S(o))(1/2)], where S(o) is the surfactant (FL-70) concentration and K* is the distribution coefficient associated with the langmuirian isotherm of the apparent effective mass against S(o). We applied this to study the surfactant effect on the retention behaviors of polystyrene (PS) latex beads of 170-500 nm in diameter. As a result, an empirical relation was found to hold between v*(b,o) and d(o) (estimated from R(o) at S(o) = 0) as v*(b,o) - v*(o,o)[1 - (d(c)/d(o))], where v*(o,o) is the asymptotic value of v*(b,o) in the vanishing d(c)/d(o) limit and d(c) is the cutoff value at which v*(b,o) would vanish. According to the present approach, the no-slip BC (v*(b,o) = 0) was predicted to recover when d(o) ∼ d(c), and the boundary slip effect could be significant for S(o) ≤ 0.05%, particularly for large latex beads.

A new strategy for pressed powder eye shadow analysis: Allergenic metal ion content and particle size distribution

Nine cheap eye shadow products were analyzed through graphite furnace atomic absorption spectrometry (GF-AAS) to quantify their Cr, Co and Ni contents, all known to be skin sensitizers. In many cases, the concentrations were higher than 1 or 5ppm (μg/g), i.e. the limits recommended in the scientific literature to minimize the risk of reaction in particularly sensitive subjects. In most cases, the concentration of Cr was higher than that of Ni and Co, up to a limit case of 150mg/g. In this particular sample, the potential amount of Cr that could be released in ionic form was determined in sweat simulating solutions by GF-AAS and confirmed through a specific spectrofluorimetric method; the results indicated the presence of approximately 80–90ppb (ng/g) of Cr3+.The water dispersible particles were isolated from the eye shadow powders through a simple solvent extraction procedure. The aqueous suspensions were then sorted through sedimentation field flow fractionation (SdFFF) and the particles sizes were calculated from experimental fractograms using theory. For the most part, the computed sizes were in the micron range, as confirmed by some SEM photographs taken on fractions collected during the separations. The SdFFF coupled off-line with the GFAAS enabled elemental characterization of pigment particles as a function of size. This finding reduces the concern that the ingredients of such makeup formulations may contain nanoparticles.

Size Selectivity in Field-Flow Fractionation: Lift Mode of Retention with Near-Wall Lift Force

A simple theoretical model for the size selectivity, S(d), in the lift mode of retention in field-flow fractionation (FFF) is developed on the basis of the near-wall lift force expression. S(d) is made up of two contributions: the flow contribution, S(d,f), arising from the variation of the flow velocity at center of particle due to a change in particle position with particle size, and a slip contribution, S(d,s), arising from the concomitant change in the extent of retardation due to the presence of a nearby channel wall. The slip contribution is minor, but not negligible, and amounts to 10-20% of the overall size selectivity. It contributes to reduce S(d) in sedimentation FFF but to enhance it in flow FFF. S(d) would steadily increase with particle size if the flow profile was linear (Couette flow). Because of the curvature of the flow profile encountered in the classical Poiseuille flow, S(d) exhibits a maximum at some specific particle size. The model predicts a significant difference in S(d) between sedimentation FFF and flow FFF, arising from the different functional dependences of the field force with particle size between these two methods. The predictions are in good agreement with the various S(d) values reported in the literature in both sedimentation and flow FFF. On the basis of the model, guidelines are given for adjusting the operating parameters (carrier flow rate and field strength) to optimize the size selectivity. Finally, it is found that S(d) generally decreases with decreasing channel thickness.

Influence of secondary preparative parameters and aging effects on PLGA particle size distribution: a sedimentation field flow fractionation investigation

Poly(lactic-co-glycolic acid) particles in the 200-400-nm size range were formulated through nanoprecipitation and solvent evaporation methods. Different concentrations of the polymer and stabilizer (Pluronic® F 68) were tested in order to identify the best conditions for making poly(lactic-co-glycolic acid) particles of suitable size, stable in time, and to be used as carriers for brain-targeting drugs. The particles with the best characteristics for delivery system design were those formulated by nanoprecipitation with an organic/water phase ratio of 2:30, a polymer concentration of 25 mg/mL, and a surfactant concentration of 0.83 mg/mL; their surface charge was reasonably negative (approximately -27 mV) and the average size of the almost monodisperse population was roughly 250 nm. Particle characterization was obtained through ζ-potential measurements, scanning electron microscope observations, and particle size distribution determinations; the latter achieved by both photon-correlation spectroscopy and sedimentation field flow fractionation. Sedimentation field flow fractionation, which is considered more reliable than photon-correlation spectroscopy in describing the possible particle size distribution modifications, was used to investigate the effects of 3 months of storage at 4 °C had on the lyophilized particles. Figure Particle size ditribution from the SdFFF and the PCS techniques.

Cancer Stem Cell Sorting from Colorectal Cancer Cell Lines by Sedimentation Field Flow Fractionation

Recently, cancer stem cells (CSCs) have been identified in many types of cancers, such as colorectal cancer (CRC). CSCs seem to be involved in initiation, growth, and tumor metastasis, as well as in radio- and chemotherapy failures. CSCs appears as new biological targets for cancer therapy, requiring the development of noninvasive cell sorting methods. In this study, we used sedimentation field flow fractionation (SdFFF) to prepare enriched populations of CSCs from eight cell lines corresponding to different CRC grades. On the basis of phenotypic and functional characterizations, "hyperlayer" elution resulted in a fraction overexpressing CSC markers (CD44, CD166, EpCAM) for all cell lines. CSCs were eluted in the last fraction for seven out of eight cell lines, but in the first for HCT116. These results suggest, according to the literature, that two different pools of CSCs exist, quiescent and activated, which can both be sorted by SdFFF. Moreover, according to CSC properties, enriched fractions are able to form colonies.

Synthesis and Size Characterization of Silica Nanospheres Using Sedimentation Field-Flow Fractionation (SdFFF)

Silica nanoparticles were synthesized by a conventional emulsion polymerization by mixing ethanol, ammonium hydroxide, water and tetra ethyl orthosilicate (TEOS). A new reaction apparatus was assembled for a large scale synthesis of silica nanospheres in the laboratory, which was designed for uniform mixing of the reactants. The apparatus was equipped with a disc type agitator with six rectangular propellers. The new apparatus allowed high reproducibility in terms of the mean size and the size distribution of the silica nanoparticles with the relative standard deviation of less than about 6%. Sedimentation field-flow fractionation (SdFFF) was employed for determination of the size distribution of the silica nanoparticles. SdFFF provided size-based separation of the silica nanoparticles, with the retention time increasing with the size. When SdFFF analysis was repeated three times for the same sample, the standard deviation was less than 4%, showing reliability of SdFFF in size measurement. SdFFF seems to provide more accurate size distribution than DLS, particularly for those having broad and multimodal size distributions. Change in the agitation speed resulted in significant change in the mean diameter of the silica nanoparticles. Agitation speed of 400 rpm in 3 L reaction vessel yielded silica particles of about 100 nm in diameter, while at 200 rpm in 1 L vessel yielded those of about 500 nm.

Colloidal mercury (Hg) distribution in soil samples by sedimentation field-flow fractionation coupled to mercury cold vapour generation atomic absorption spectroscopy

Diverse analytical techniques are available to determine the particle size distribution of potentially toxic elements in matrices of environmental interest such as soil, sediments, freshwater and groundwater. However, a single technique is often not exhaustive enough to determine both particle size distribution and element concentration. In the present work, the investigation of mercury in soil samples collected from a polluted industrial site was performed by using a new analytical approach which makes use of sedimentation field-flow fractionation (SdFFF) coupled to cold vapour generation electrothermal atomic absorption spectroscopy (CV-ETAAS). The Hg concentration in the SdFFF fractions revealed a broad distribution from about 0.1 to 1 μm, roughly following the particle size distributions, presenting a maximum at about 400-700 nm in diameter. A correlation between the concentration of Hg in the colloidal fraction and organic matter (O.M.) content in the soil samples was also found. However, this correlation is less likely to be related to Hg sorption to soil O.M. but rather to the presence of colloidal mercuric sulfide particles whose size is probably controlled by the occurrence of dissolved O.M. The presence of O.M. could have prevented the aggregation of smaller particles, leading to an accumulation of mercuric sulfides in the colloidal fraction. In this respect, particle size distribution of soil samples can help to understand the role played by colloidal particles in mobilising mercury (also as insoluble compounds) and provide a significant contribution in determining the environmental impact of this toxic element.

Overcoming challenges in analysis of polydisperse metal-containing nanoparticles by single particle inductively coupled plasma mass spectrometry

Detection and sizing of metal-containing engineered nanoparticles (ENPs) was achieved at concentrations predicted for environmental samples (part-per trillion levels) using single particle inductively coupled plasma mass spectrometry (SP-ICP-MS). Silver nanowires, titanium dioxide and cerium oxide nanoparticles were found to be detectable by this technique, while zinc oxide dissolved too quickly for analysis at these concentrations. In addition to the potential dissolution of particles, other considerations affecting ENP analysis include: instrumental background, mass interferences, percent metal in a nanoparticle, and isotopic abundance of the analyte element. Sizing of these metal-containing nanoparticles was done by correlating ICP-MS response (pulses) from ENPs entering the plasma to mass of metal in dissolved standards. The resulting particle size distributions compared well with results from sedimentation field-flow fractionation. Coincidence in ENP pulses may be difficult to detect in the broad size distributions that arise from polydisperse samples. Comparison of data obtained by combination of multiple analyses of dilute solutions to single analyses of higher concentration allowed discrimination between coincidence and polydispersity. The ratio of ENP pulse detections to the total number of readings during analysis was optimized at 2.5% or less to minimize coincident pulses while still allowing definition of a size distribution.

마이크로 입자의 습식 수집 및 분획 장치 개발

본 연구에서는 마이크로 크기의 입자(안료, 대기입자, 등)를 수집과 동시에 물에 분산된 상태로 분획가능한 장치를 개발하였다. 이 장치에서는 물을 넣은 입자수집용 실린더 세 개를 직렬로 연결하여 공기를 통과하게 하였다. 그럼으로써 크기가 다른 입자들이 다른 실린더에 모아지도록 하였고, 수집과 동시에 분획이 가능하도록 하였다. 이 장치의 중요한 특징은 막필터(membrane filter)를 사용하지 않는다는 점이다. 즉, 필터를 사용하지 않고 마이크로 크기의 입자를 직접 시료 수집용 실린더에 수집되도록 하였다. 또한 수집된 시료는 수집되는 동안 물에 분산된 상태로 장치 내에서 동시분획이 가능하다. 이 장치를 에어본 입자의 수집에 응용하였다. 수집된 에어본 입자는 침강 장-흐름 분획법(sedimentation field-flow fractionation, SdFFF)을 이용하여 입자 크기별로 분획하였고, 광학현미경(optical microscopy, OM)을 이용하여 입자 크기와 모양을 관찰하였다. 또한 AAS와 ICP-AES를 이용하여 에어본 입자들의 조성을 분석하였다. 본 연구를 통하여 개발한 입자 수집 및 분획장치는 다양한 종류의 입자의 수집 및 크기분포 분석에 활용이 가능할 것으로 기대한다. In this study, a wet-sampler was developed for collection in water and simultaneous fractionation of micron-sized particles (e.g., pigment, airborne, etc.). In this new device, three cylinders (partially filled with water) through which air was forced to pass for sample collection are connected in a series. Particles of different sizes are collected in different cylinders, allowing simultaneous fractionation. An uniqueness of this new device is that it does not use any membrane filter. Microparticles are collected directly in cylinders filled partially with water. Also the particles are simultaneously fractionated within this device while they are being collected. The new device was employed for collection of airborne particles. The collected airborne particles were fractionated by using sedimentation field-flow fractionation, SdFFF), and observed with an optical microscopy (OM) for size and shape analysis. Also AAS and ICP-AES were used for composition analysis of the airborne particles. It is expected that the new device is applicable to collection and analysis of size distribution of various types of microparticles.