Kayori Takahashi

Reliable size determination of nanoparticles using dynamic light scattering method for in vitro toxicology assessment.

Dynamic light scattering (DLS) is widely used for the evaluation of the particle size in the toxicity assessment of nanoparticles. However, the many types of DLS instruments and analytical procedures sometimes give different apparent sizes of particles and make it complicated to understand the size dependence on particles for the toxicity assay. In this study, we established an evaluation method of secondary nanoparticle sizes using a DLS analysis. First, we established a practical method for determining size with an appropriate evaluation of uncertainties. This proposed method could be a universal protocol for toxicity assessment that would allow researchers to achieve some degree of concordance on the size of nanoparticles for an assessment. Second, we investigated the processes associated with particles in suspension by examining the changes in the size and the light scattering intensity of secondary nanoparticles during an in vitro toxicity assessment, since the transport mode of particles to cells is significant in understanding in vitro nano-toxicity. In this study, these two points were investigated on TiO(2) nanoparticles suspension as an example. The secondary particles of TiO(2) with a light scattering intensity-averaged diameter (d(l)) of 150-250 nm were characterized with appropriate uncertainties. The sizes were found to be comparable with values determined using other analytical procedures and other instruments. It is suggested that d(l) could be an effective size parameter for toxicity assessments. Furthermore, TiO(2) secondary nanoparticle suspensions are well dispersed with slow gravity settling, no agglomeration, with the diffusion process as the primary transport mode of particles to cells.

Quantitative comparison of a corona-charged aerosol detector and an evaporative light-scattering detector for the analysis of a synthetic polymer by supercritical fluid chromatography

A corona-charged aerosol detector (CAD) was developed to improve the sensitivity, reproducibility and quantitativeness of detection as compared to evaporative light-scattering detector (ELSD) for liquid chromatography. Our laboratory used the corona CAD as a detector for supercritical fluid chromatography (SFC) and evaluated its performance compared to the ELSD by using a certified reference material of poly(ethylene glycol) (PEG) and a well-defined equimass mixture of uniform PEG oligomers. The corona CAD was able to detect a 10 times more dilute solution of uniform oligomers compared to the ELSD. Although the original data of molecular mass by ELSD was 4.6% smaller than the certified value of PEG 1000, molecular mass distribution obtained by corona CAD was virtually almost the same as the certified value without any calibrations.

Dispersion characteristics of various metal oxide secondary nanoparticles in culture medium for in vitro toxicology assessment.

The aim of this study is to characterize the dispersion characteristics of various metal oxide nanoparticles and secondary nanoparticle formation in culture medium. Many studies have already investigated the in vitro toxicities of various metal oxide nanoparticles; however, there have been few discussions about the particle transport mode to cells during a period of toxicity assessment. The particle transport mode would strongly affect the amount of uptake by cells; therefore, estimation of the transport mode for various metal oxide particles is important. Fourteen different metal oxide nanoparticle dispersions in a culture medium were examined. The sizes of the secondary nanoparticles were observed to be larger than 100 nm by dynamic light scattering (DLS). According to Stokes law and the Stokes-Einstein assumption, pure metal oxide particles with such sizes should gravitationally settle faster than diffusion processes; however, the secondary metal oxide particles examined in this study exhibited unexpectedly slower gravitational settling rates. The slow gravitational settling kinetics of particles was estimated to be caused by the inclusion of protein into the secondary nanoparticles, which resulted in lower densities than the pure metal oxide particles. The ratios of metal oxide to protein in secondary particles could be affected by the protein adsorption ability of the corresponding metal oxide primary particles. To the best of our knowledge, it was clarified for the first time that stably dispersed secondary metal oxide nanoparticles with slow gravitational settling kinetics are induced by secondary nanoparticles consisting of small amounts of metal oxide particles and large amounts of protein, which results in lower particle densities than the pure metal oxide particles. The estimation of particle dynamics in culture medium using this method would be significant to recognize the inherent toxicity of nanoparticles.

Accurate Size and Size-Distribution Determination of Polystyrene Latex Nanoparticles in Aqueous Medium Using Dynamic Light Scattering and Asymmetrical Flow Field Flow Fractionation with Multi-Angle Light Scattering

Accurate determination of the intensity-average diameter of polystyrene latex (PS-latex) by dynamic light scattering (DLS) was carried out through extrapolation of both the concentration of PS-latex and the observed scattering angle. Intensity-average diameter and size distribution were reliably determined by asymmetric flow field flow fractionation (AFFFF) using multi-angle light scattering (MALS) with consideration of band broadening in AFFFF separation. The intensity-average diameter determined by DLS and AFFFF-MALS agreed well within the estimated uncertainties, although the size distribution of PS-latex determined by DLS was less reliable in comparison with that determined by AFFFF-MALS.

Size effect on UV-Vis absorption properties of colloidal C60 particles in water

An obvious size effect of colloidal C(60) particles on UV-Vis absorption spectra was found after size fractionation of colloidal C(60) particles in water using the Field Flow Fractionation method.

Polymer analysis by supercritical fluid chromatography.

Synthetic polymers always have distributions of the degrees of polymerization. Supercritical fluid chromatography (SFC) is almost only one technique to be able to separate pure polymer that has exactly unique degree of polymerization. For typical examples of SFC separation of polymers, four kinds of polymers: polystyrene, poly(ethylene glycol), some nonionic surfactants, and polyprenol are described about the conditions of SFC. Separated pure polymers which have no molecular weight distribution, are called uniform polymer. Uniform polymers are really useful to calibrate or validate various detectors. Here, the applications of calibrating ultraviolet detector, evaporative light scattering detector, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry are summarized. These detectors have strong molecular weight dependence of their sensitivity coefficient, in contrast to the good linearity of sample concentration.

Evaluation of a condensation nucleation light scattering detector for the analysis of synthetic polymer by supercritical fluid chromatography

A condensation nucleation light scattering detector (CNLSD) was adapted for use as a detector for supercritical fluid chromatography. The performance of the CNLSD was evaluated and compared to evaporative light-scattering detector (ELSD) using a well-defined equimass mixture of uniform poly(ethylene glycol) oligomers and a certified reference material of poly(ethylene glycol) 1000. The CNLSD was able to detect a 10 times less concentrated solution of uniform oligomers compared to the ELSD. The quantitativeness of CNLSD was high enough to obtain the molecular mass distribution of poly(ethylene glycol) 1000 without any calibrations; on the other hand, the original data measured by ELSD was about 4% smaller than the certified value of poly(ethylene glycol) 1000. The CNLSD was suitable for supercritical fluid chromatography as a mass concentration detector.

Calibration of an evaporative light-scattering detector as a mass detector for supercritical fluid chromatography by using uniform Poly(ethylene glycol) oligomers.

The quantitativeness of an evaporative light-scattering detector (ELSD) for supercritical fluid chromatography (SFC) was evaluated by using an equimass mixture of uniform poly(ethylene glycol) (PEG) oligomers. Uniform oligomers, in which all molecules have an identical molecular mass, are useful for the accurate calibration of detectors. We calibrated the SFC-ELSD system for various concentrations and molecular masses by using an equimass mixture of PEG oligomers. ELSD not only showed a good linear response to the injected concentration over a wide concentration range, from 10(-4) to 10(-1)g/mL, but also showed a strong dependence on the molecular mass of the solute. By using chromatograms of the equimass mixture of uniform oligomers to calibrate SFC-ELSD, it was possible to determine exact values of not only the average mass but also the molecular-mass distribution for a PEG 1540 sample. The average molecular mass was shifted to a higher value by several percentage points after calibration of the ELSD.

Certification and uncertainty evaluation of the certified reference materials of poly(ethylene glycol) for molecular mass fractions by using supercritical fluid chromatography

AbstractPoly(ethylene glycol) (PEG) is a useful water-soluble polymer that has attracted considerable interest in medical and biological science applications as well as in polymer physics. Through the use of a well-calibrated evaporative light-scattering detector coupled with high performance supercritical fluid chromatography, we are able to determine exactly not only the average mass but also all of the molecular mass fractions of PEG samples needed for certified reference materials issued by the National Metrology Institute of Japan. In addition, experimental uncertainty was determined in accordance with the Guide to the expression of uncertainty in measurement (GUM). This reference material can be used to calibrate measuring instruments, to control measurement precision, and to confirm the validity of measurement methods when determining molecular mass distributions and average molecular masses. Especially, it is suitable for calibration against both masses and intensities for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. FigureComparison between the molecular mass fractions of PEG 1000 before calibration (si) (○) and after calibration (wi) (⧫). The error bar shows the expanded uncertainty of k = 2 of each mass fraction

Recent activity of international comparison for nanoparticle size measurement

Nanoparticle sizing is the most fundamental measurement for producing nanomaterials, evaluation of nanostructure, and the risk assessment of nanomaterials for human health. Dynamic light scattering (DLS) is widely used as a useful and convenient technique for determining nanoparticle size in liquid; however, the precision of this technique has been unclear. Some international comparisons are now in progress to verify the measurement accuracy of nanoparticle sizing, as a typical example of Asia Pacific Metrology Programme Supplementary Comparison. In this study, we evaluated the precision of DLS technique for nanoparticle sizing and estimated the uncertainty of the DLS data for polystyrene latex suspensions. The extrapolations of apparent diffusion coefficients to infinite dilution and to lower angles yielded more precise values than those obtained at one angle and one concentration. The extrapolated particle size measured by DLS was compared to the size determined by differential mobility analyzer (DMA), atomic force microscopy (AFM), and scanning electron microscopy (SEM). Before the comparison, the intensity-averaged size measured by DLS was recalculated to the number-averaged size, and the thickness of water layer attaching on the surface of particles were added into uncertainty of particle sizing by DLS. After the recalculation, the consistent values of mean particle diameter were obtained between those by DLS and by DMA, AFM, and SEM within the estimated uncertainties.