Flow Field–Flow Fractionation for size analysis and characterization of nanoparticles for applications in Life Sciences

Nanotechnologies are rapidly expanding because of the opportunities that the new materials offer in many areas such as the manufacturing industry, food production, processing and preservation, and in the pharmaceutical and cosmetic industry. Size distribution of the nanoparticles determines their properties and is a fundamental parameter that needs to be monitored from the small-scale synthesis up to the bulk production and quality control of nanotech products on the market. A consequence of the increasing number of applications of nanomaterial is that the EU regulatory authorities are introducing the obligation for companies that make use of nanomaterials to acquire analytical platforms for the assessment of the size parameters of the nanomaterials. In this work, Asymmetrical Flow Field-Flow Fractionation (AF4) and Hollow Fiber F4 (HF5), hyphenated with Multiangle Light Scattering (MALS) are presented as tools for a deep functional characterization of nanoparticles. In particular, it is demonstrated the applicability of AF4-MALS for the characterization of liposomes in a wide series of mediums. Afterwards the technique is used to explore the functional features of a liposomal drug vector in terms of its biological and physical interaction with blood serum components: a comprehensive approach to understand the behavior of lipid vesicles in terms of drug release and fusion/interaction with other biological species is described, together with weaknesses and strength of the method. Afterwards the size characterization, size stability, and conjugation of azidothymidine drug molecules with a new generation of metastable drug vectors, the Metal Organic Frameworks, is discussed. Lastly, it is shown the applicability of HF5-ICP-MS for the rapid screening of samples of relevant nanorisk: rather than a deep and comprehensive characterization it this time shown a quick and smart methodology that within few steps provides qualitative information on the content of metallic nanoparticles in tattoo ink samples.

Nanoparticular iron complex drugs for parenteral administration - physicochemical characterization, biological distribution and pharmacological safety

Iron deficiency is the most common deficiency disease worldwide with many patients who require intravenous iron. Within the last years new kind of parenteral iron complexes as well as generic preparations entered the market. There is a high demand for methods clarifying benefit to risk profiles of old and new iron complexes. It is also necessary to disclose interchangeability between originator and intended copies to avoid severe anaphylactic and anaphylactoid side reaction and assure equivalence of therapeutic effect.rnrnThe investigations presented in this work include physicochemical characterization of nine different parenteral iron containing non-biological complex drugs. rnWe developed an in-vitro assay, which allows the quantification of labile iron in the different complexes and thus it is a useful tool to estimate the pharmaclogical safety regarding iron related adverse drug events. This assay additionally allowed the estimation of complex stability by evaluation of degradation kinetics at the applied conditions.rnrnAn in-ovo study was performed to additionally compare different complexes in respect to body distribution. This in combination with complex stability information allowed the risk estimation of potential local acute and chronic reactions to iron overload.rnrnInformation obtained by the combination of the methods within this work are helpful to estimate the safety and efficacy profile of different iron containing non-biological complex drugs. rnrnPhysicochemical differences between the complexes were demonstrated in respect to size of the inorganic fraction, size and size distribution of the complete particles, structure of the inorganic iron fraction, morphology of the complexes and charge of the complexes. And furthermore significant differences in the biological behavior of different complexes were demonstrated. rnrnThe combination of complex stability and biodistribution as well as the combination of structure, size and stability represent helpful tools for the physicochemical characterization of iron containing non-biological complex drugs and for the estimation of pharmacological safety. This work thus represents an up to date summary of some relevant methods for the characterization of intravenous iron complex drugs in respect to pharmaceutical quality, pharmacological safety and aspects of efficacy. rnrnProspectively, it is worthwhile that the methods within this work will contribute to the development and/or characterization of iron containing nanoparticular formulations with beneficial efficacy and safety profiles.rn

Ecological distribution of semi-demersal fishes in space and time on the shelf of Antalya Gulf

In this study we provide a baseline data on semidemersal fish assemblages and biology in a heterogeneous and yet less studied portion of the shelf of Antalya Gulf. The distribution of fish abundance in three transects subjected to different fisheries regulations (fishery vs non fishery areas), and including depths of 10, 25, 75, 125, 200 m, was studied between May 2014 and February 2015 in representative months of winter, spring, summer and autumn seasons. A total of 76 fish species belonging to 40 families was collected and semidemersal species distribution was analyzed in comparison with the whole community. Spatial distribution of fish was driven mainly by depth and two main assemblages were observed: shallow waters (10-25; 75 m) and deep waters (125-200 m). Significant differences among transects were found for the whole community but not for the semidemersal species. Analysis showed that this was due to a strong relation of these species with local environmental characteristics rather than to a different fishing pressure over transects. Firstly all species distribute according to the bathymetrical gradient and secondly to the bottom type structure. Semidemersal species were then found more related to zooplankton and suspended matter availability. The main morphological characteristics, sex and size distribution of the target semidemersal species Spicara smaris (Linnaeus, 1758), Saurida undosquamis (Richardson, 1848), Pagellus acarne (Risso, 1827) were also investigated.

The role of seasonality of mineral dust concentration and size on glacial/interglacial dust changes in the EPICA Dronning Maud Land ice core

We present a record of particulate dust concentration and size distribution in subannual resolution measured on the European Project for Ice Coring in Antarctica (EPICA) Dronning Maud Land (EDML) ice core drilled in the Atlantic sector of the East Antarctic plateau. The record reaches from present day back to the penultimate glacial until 145,000 years B.P. with subannual resolution from 60,000 years B.P. to the present. Mean dust concentrations are a factor of 46 higher during the glacial (~850–4600 ng/mL) compared to the Holocene (~16–112 ng/mL) with slightly smaller dust particles during the glacial comparedto the Holocene and with an absolute minimum in the dust size at 16,000 years B.P. The changes in dust concentration are mainly attributed to changes in source conditions in southern South America. An increase in the modal value of the dust size suggests that at 16,000 years B.P. a major change in atmospheric circulation apparently allowed more direct transport of dust particles to the EDML drill site. We find a clear in-phase relation of the seasonal variation in dust mass concentration and dust size during the glacial (r(conc,size) = 0.8) but no clear phase relationship during the Holocene (0

Mineralogical and grain size analysis of ODP Hole 119-745B from the Southern Ocean

The upper Miocene to Pleistocene sediments recovered at ODP Sites 745 and 746 in the Australian-Antarctic Basin are characterized by cyclic facies changes. Sedimentological investigations of a detailed Quaternary section reveal that facies A is dominated by a high content of siliceous microfossils, a relatively low terrigenous sediment content, an ice-rafted component, low concentrations of fine sediment particles, and a relatively high smectite content. This facies corresponds to interglacial sedimentary conditions. Facies B, in contrast, is characteristic of glacial conditions and is dominated by a large amount of terrigenous material and a smaller opaline component. There is also a prominent ice-rafted component. The microfossils commonly are reworked and broken. The clay mineral assemblages show higher proportions of glacially derived illite and chlorite. A combination of four different processes, attributed to glacial-interglacial cycles, was responsible for the cyclic facies changes during Quaternary time: transport by gravity, ice, and current and changes in primary productivity. Of great importance was the movement of the grounding line of the ice shelves, which directly influenced the intensity of ice rafting and of gravitational sediment transport to the deep sea. The extension of the ice shelves was also responsible for the generation of cold and erosive Antarctic Bottom Water, which controlled the grain-size distribution, particularly of the fine fraction, in the investigated area.

Clay mineraology and grain size composition of surface sediments from South Pacific Ocean

The reconstruction of low-latitude ocean-atmosphere interactions is one of the major issues of (paleo-)environmental studies. The trade winds, extending over 20° to 30° of latitude in both hemispheres, between the subtropical highs and the intertropical convergence zone, are major components of the atmospheric circulation and little is known about their long-term variability on geological time-scales, in particular in the Pacific sector. We present the modern spatial pattern of eolian-derived marine sediments in the eastern equatorial and subtropical Pacific (10°N to 25°S) as a reference data set for the interpretation of SE Pacific paleo-dust records. The terrigenous silt and clay fractions of 75 surface sediment samples have been investigated for their grain-size distribution and clay-mineral compositions, respectively, to identify their provenances and transport agents. Dust delivered to the southeast Pacific from the semi- to hyper-arid areas of Peru and Chile is rather fine-grained (4-8 µm) due to low-level transport within the southeast trade winds. Nevertheless, wind is the dominant transport agent and eolian material is the dominant terrigenous component west of the Peru-Chile Trench south of ~ 5°S. Grain-size distributions alone are insufficient to identify the eolian signal in marine sediments due to authigenic particle formation on the sub-oceanic ridges and abundant volcanic glass around the Galapagos Islands. Together with the clay-mineral compositions of the clay fraction, we have identified the dust lobe extending from the coasts of Peru and Chile onto Galapagos Rise as well as across the equator into the doldrums. Illite is a very useful parameter to identify source areas of dust in this smectite-dominated study area.

Grain size composition and age of ODP Hole 167-1017E sediments

Siliciclastic sedimentation at Ocean Drilling Program Site 1017 on the southern slope of the Santa Lucia Bank, central California margin, responded closely to oceanographic and climatic change over the past ~130 ka. Variation in mean grain-size and sediment sorting within the ~25-m-thick succession from Hole 1017E show Milankovitch-band to submillenial-scale variation. Mean grain size of the "sortable silt" fraction (10-63 µm) ranges from 17.6 to 33.9 µm (average 24.8 µm) and is inversely correlated with the degree of sorting. Much of the sediment has a bimodal or trimodal grain-size distribution that is composed of distinct fine silt, coarse silt to fine sand, and clay-size components. The position of the mode and the sorting of each component changes through the succession, but the primary variation is in the presence or abundance of the coarse silt fraction that controls the overall mean grain size and sorting of the sample. The occurrence of the best-sorted, finest grained sediment at high stands of sea level (Holocene, marine isotope Substages 5c and 5e) reflect the linkage between global climate and the sedimentary record at Site 1017 and suggest that the efficiency of off-shelf transport is a key control of sedimentation on the Santa Lucia Slope. It is not clear what proportion of the variation in grain size and sorting may also be caused by variations in bottom current strength and in situ hydrodynamic sorting.

Sediment organic matter, grain size, and results of prediction models from northern North Atlantic and Arctic Seas

Sediment samples and hydrographic conditions were studied at 28 stations around Iceland. At these sites, Conductivity-Temperature-Depth (CTD) casts were conducted to collect hydrographic data and multicorer casts were conductd to collect data on sediment characteristics including grain size distribution, carbon and nitrogen concentration, and chloroplastic pigment concentration. A total of 14 environmental predictors were used to model sediment characteristics around Iceland on regional geographic space. For these, two approaches were used: Multivariate Adaptation Regression Splines (MARS) and randomForest regression models. RandomForest outperformed MARS in predicting grain size distribution. MARS models had a greater tendency to over- and underpredict sediment values in areas outside the environmental envelope defined by the training dataset. We provide first GIS layers on sediment characteristics around Iceland, that can be used as predictors in future models. Although models performed well, more samples, especially from the shelf areas, will be needed to improve the models in future.

Sedimentverteilung im Seegebiet bei Bokniseck, Eckernförder Bucht, westliche Ostsee (sediment distribution at Bokniseck, western Baltic Sea; map)

A detailed topographic survey was carried out in the "Hausgarten"-area of the Joint Research Programm 95 of the University of Kiel by the Deutsches Hydrographisches Institut. Based on this information a sediment distribution map was constructed. A horizontal section extending from 0 to 27 m of water depth was investigated showing the distribution of pebbles and boulders, of algal growth, and exposed areas of glacial marl; the grain size distribution was determined for the various sediment types.

Vertical distribution of suspended aggregates at time series station DYNAPROC

Day/night variations in the size distribution of the particulate matter >0.15 mm (PM) were studied in May 1995 during the DYNAPROC time-series cruise in the northwestern Mediterranean Sea. Data on vertical distributions of PM (>0.15 mm) and zooplankton were collected with the Underwater Video Profiler (UVP). The comparisons of the UVP data with plankton net data and POC data from water bottles indicated that more than 97% of the particles detected by the UVP were non-living particles (0.15 mm) and that the PM contributed 4-34% of the total dry weight measured on GF/F filters. Comparison of seven pairs of day and night vertical profiles performed during the cruise showed that in the upper 800 m, the mean size and the volume of particles was higher at night than during the day. During the night, the integrated volume of the PM increased on average by 32±20%. This increase corresponded to a shift of smaller size classes (<0.5 mm) towards the larger ones (>0.5 mm). During the day, the pattern was reversed, and the quantity of PM >0.5 mm decreased. During the study period, the standing stock of PM (60-800 m) decreased from 7.5 to less than 2 g m?2 but the diel variations persisted, except for two short periods in the superficial layer following a wind event. The cyclic feeding activity induced by the diel vertical migration of zooplankton could be the best candidate to explain the observed diel fluctuations in the size classes of PM in the water column. However, our results also suggest that in the upper layer additional driving forces such as the increase of the level of turbulence after a wind event or the modification of the zoo- and phytoplankton community can influence the PM temporal evolution.

Grain size investigations of sediments off the coast of Istria

Composition, grain-size distribution, and areal extent of Recent sediments from the Northern Adriatic Sea along the Istrian coast have been studied. Thirty one stations in four sections vertical to the coast were investigated; for comparison 58 samples from five small bays were also analyzed. Biogenic carbonate sediments are deposited on the shallow North Adriatic shelf off the Istrian coast. Only at a greater distance from the coast are these carbonate sediments being mixed with siliceous material brought in by the Alpine rivers Po, Adige, and Brenta. Graphical analysis of grain-size distribution curves shows a sediment composition of normally three, and only in the most seaward area, of four major constituents. Constituent 1 represents the washed-in terrestrial material of clay size (Terra Rossa) from the Istrian coastal area. Constituent 2 consists of fine to medium sand. Constituent 3 contains the heterogeneous biogenic material. Crushing by organisms and by sediment eaters reduces the coarse biogenic material into small pieces generating constituent 2. Between these two constituents there is a dynamic equilibrium. Depending upon where the equilibrium is, between the extremes of production and crushing, the resulting constituent 2 is finer or coarser. Constituent 4 is composed of the fine sandy material from the Alpine rivers. In the most seaward area constituents 2 and 4 are mixed. The total carbonate content of the samples depends on the distance from the coast. In the near coastal area in high energy environments, the carbonate content is about 80 %. At a distance of 2 to 3 km from the coast there is a carbonate minimum because of the higher rate of sedimentation of clay-sized terrestrial, noncarbonate material at extremely low energy environments. In an area between 5 and 20 km off the coast, the carbonate content is about 75 %. More than 20 km from the shore, the carbonate content diminishes rapidly to values of about 30 % through mixing with siliceous material from the Alpine rivers. The carbonate content of the individual fractions increases with increasing grain-size to a maximum of about 90 % within the coarse sand fractions. Beyond 20 km from the coast the samples show a carbonate minimum of about 13 % within the sand-size classes from 1.5 to 0.7 zeta¬? through mixing with siliceous material from the alpine rivers. By means of grain-size distribution and carbonate content, four sediment zones parallel to the coast were separated. Genetically they are closely connected with the zonation of the benthic fauna. Two cores show a characteristic vertical distribution of the sediment. The surface zone is inversely graded, that means the coarse fractions are at the top and the fine fractions are at the bottom. This is the effect of crushing of the biogenic material produced at the surface by predatory organisms and by sediment eaters. lt is proposed that at a depth of about 30 cm a chemical solution process begins which leads to diminution of the original sediment from a fine to medium sand to a silt. The carbonate content decreases from about 75 % at the surface to 65 % at a depth of 100 cm. The increase of the noncarbonate components by 10 % corresponds to a decrease in the initial amount of sediment (CaC03=75 %) by roughly 30 % through solution. With increasing depth the carbonate content of the individual fractions becomes more and more uniform. At the surface the variation is from 30 % to 90 %, at the bottom it varies only between 50 % and 75 %. Comparable investigations of small-bay sediments showed a c1ear dependence of sediment/faunal zonation from the energy of the environment. The investigations show that the composition and three-dimensional distribution of the Istrian coastal sediments can not be predicted only from one or a few measurable factors. Sedimentation and syngenetic changes must be considered as a complex interaction between external factors and the actions of producing and destroying organisms that are in dynamic equilibrium. The results obtained from investigations of these recent sediments may be of value for interpreting fossil sediments only with strong limitations.

Bulk geochemistry and grain-size composition of surface sediments from the SW Atlantic

Surface sediments from the South American continental margin surrounding tbe Argentine Basin were studied with respect to bulk geochemistry (Caeo) and C ) and grain-size composition (sand/silt/clay relation and terrigenous silt grain-size distribution). The grain-size distributions of the terrigenous silt fraction were unmixed into three end members (EMs), using an end-member modelling algorithm. Three unimodal EMs appear to satisfactorily explain the variations in the data set of the grain-size distributions ofterrigenous silt. The EMs are related to sediment supply by rivers, downslope transport, winnowing, dispersal and re-deposition by currents. The bulk geochemical composition was used to trace the distribution of prominent water masses within the vertical profile. The sediments of the eastern South American continental margin are generally divided into a coarse-grained and carbonate-depleted southwestern part, and a finer-grained and carbonate-rich northeastern part. The transition of both environments is located at the position of the Brazil Malvinas Confluence (BMC). The sediments below the confluence mixing zone of the Malvinas and Brazil Currents and its extensions are characterised by high concentrations of organic carbon, low carbonate contents and high proportions of the intennediate grain-size end member. Tracing these properties, the BMC emerges as a distinct north-south striking feature centered at 52-54°W crossing the continental margin diagonally. Adjacent to this prominent feature in the southwest, the direct detrital sediment discharge of the Rio de la Plata is clearly recognised by a downslope tongue of sand and high proportions of the coarsest EM. A similar coarse grain-size composition extends further south along the continental slope. However, it displays bener sorting due to intense winnowing by the vigorous Malvinas Current. Fine-grained sedimentary deposition zones are located at the southwestern deeper part of the Rio Grande Rise and the southern abyssal Brazil Basin, both within the AABW domain. Less conspicuous winnowing/accumulation panerns are indicated north of the La Plata within the NADW level according to the continental margin topography. We demonstrate that combined bulk geochemical and grain-size properties of surface sediments, unmixed with an end-member algorithm, provide a powerful tool to reconstruct the complex interplay of sedimentology and oceanography along a time slice.

Coccoliths and carbonate grain-size compositions in sediment samples from the Mid-Atlantic Ridge

This study presents a differentiated carbonate budget for marine surface sediments from the Mid-Atlantic Ridge of the South Atlantic, with results based on carbonate grain-size composition. Upon separation into sand, silt, and clay sub-fractions, the silt grain-size distribution was measured using a SediGraph 5100. We found regionally characteristic grain-size distributions with an overall minimum at 8 µm equivalent spherical diameter (ESD). SEM observations reveal that the coarse particles (>8 µm ESD) are attributed to planktic foraminifers and their fragments, and the fine particles (<8 µm ESD) to coccoliths. On the basis of this division, the regional variation of the contribution of foraminifers and coccoliths to the carbonate budget of the sediments are calculated. Foraminifer carbonate dominates the sediments in mesotropic regions whereas coccoliths contribute most carbonate in oligotrophic regions. The grain size of the coccolith share is constant over water depth, indicating a lower susceptibility for carbonate dissolution compared to foraminifers. Finally, the characteristic grain-size distribution in fine silt (<8 µm ESD) is set into context with the coccolith assemblage counted and biometrically measured using a SEM. The coccoliths present in the silt fraction are predominantly large species (length > 4 µm). Smaller species (length < 4 µm) belong to the clay fraction (<2 µm ESD). The average length of most frequent coccolith species is connected to prominent peaks in grain-size distributions (ESD) with a shape factor. The area below Gaussian distributions fitted to these peaks is suggested as a way to quantitatively estimate the carbonate contribution of single coccolith species more precisely compared to conventional volume estimates. The quantitative division of carbonate into the fraction produced by coccoliths and that secreted by foraminifers enables a more precise estimate for source/sink relations of consumed and released CO2 in the carbon cycle. The allocation of coccolith length and grain size (ESD) suggests size windows for the separation or accumulation of distinct coccolith species in investigations that depend on non to slightly-mixed signals (e.g., isotopic studies).

Grain size and XRD analysis on Site 162-983

The grain size of deep-sea sediments provides an apparently simple proxy for current speed. However, grain size-based proxies may be ambiguous when the size distribution reflects a combination of processes, with current sorting only one of them. In particular, such sediment mixing hinders reconstruction of deep circulation changes associated with ice-rafting events in the glacial North Atlantic because variable ice-rafted detritus (IRD) input may falsely suggest current speed changes. Inverse modeling has been suggested as a way to overcome this problem. However, this approach requires high-precision size measurements that register small changes in the size distribution. Here we show that such data can be obtained using electrosensing and laser diffraction techniques, despite issues previously raised on the low precision of electrosensing methods and potential grain shape effects on laser diffraction. Down-core size patterns obtained from a sediment core from the North Atlantic are similar for both techniques, reinforcing the conclusion that both techniques yield comparable results. However, IRD input leads to a coarsening that spuriously suggests faster current speed. We show that this IRD influence can be accounted for using inverse modeling as long as wide size spectra are taken into account. This yields current speed variations that are in agreement with other proxies. Our experiments thus show that for current speed reconstruction, the choice of instrument is subordinate to a proper recognition of the various processes that determine the size distribution and that by using inverse modeling meaningful current speed reconstructions can be obtained from mixed sediments.

Abundance and size composition of ctenophore Mnemiopsis leidyi population in the Caspian Sea on 2001-06-24 to 2001-07-01

Abundance and size distribution of ctenophore Mnemiopsis leidyi in different parts of the Caspian Sea were studied in summer 2001 in relation to environmental conditions. In general, principal differences were found in M. leidyi abundance and population reproduction activity in northern-, middle- and southern Caspian waters. Ctenophore was practically absent in the northern Caspian. In the west of the middle Caspian Sea it penetrated far to the north demonstrating low reproduction activity. In the east the first single comb jellies were pointed out only in the most south of the region. In the warmest and most productive southern part of the Caspian Sea several zones of M. leidyi active breeding were found with total abundance exceeding 6000 #/m**2. Breeding activity and abundance of ctenophores increased here from the east to the west exceeding maximum values along the western coast of the southern Caspian Sea in regions of intensive sprat catching. Dependence of M. leidyi population development on temperature conditions was mentioned. On the base of remote sensed surface temperature, chlorophyll, and suspended mater distribution analysis possible ctenophore settling mechanisms by mesoscale dynamic structures were examined. Practical applications of obtained results are discussed for using effective biological methods to prevent catastrophic consequences of M. leidyi invasion to the Caspian Sea.