Versailles project on advanced materials and standards (VAMAS) interlaboratory study on measuring the number concentration of colloidal gold nanoparticles.

We describe the outcome of a large international interlaboratory study of the measurement of particle number concentration of colloidal nanoparticles, project 10 of the technical working area 34, "Nanoparticle Populations" of the Versailles Project on Advanced Materials and Standards (VAMAS). A total of 50 laboratories delivered results for the number concentration of 30 nm gold colloidal nanoparticles measured using particle tracking analysis (PTA), single particle inductively coupled plasma mass spectrometry (spICP-MS), ultraviolet-visible (UV-Vis) light spectroscopy, centrifugal liquid sedimentation (CLS) and small angle X-ray scattering (SAXS). The study provides quantitative data to evaluate the repeatability of these methods and their reproducibility in the measurement of number concentration of model nanoparticle systems following a common measurement protocol. We find that the population-averaging methods of SAXS, CLS and UV-Vis have high measurement repeatability and reproducibility, with between-labs variability of 2.6%, 11% and 1.4% respectively. However, results may be significantly biased for reasons including inaccurate material properties whose values are used to compute the number concentration. Particle-counting method results are less reproducibile than population-averaging methods, with measured between-labs variability of 68% and 46% for PTA and spICP-MS respectively. This study provides the stakeholder community with important comparative data to underpin measurement reproducibility and method validation for number concentration of nanoparticles.

An Interlaboratory Comparison on the Characterization of a Sub-micrometer Polydisperse Particle Dispersion.

The measurement of polydisperse protein aggregates and particles in biotherapeutics remains a challenge, especially for particles with diameters of ≈ 1 µm and below (sub-micrometer). This paper describes an interlaboratory comparison with the goal of assessing the measurement variability for the characterization of a sub-micrometer polydisperse particle dispersion composed of five sub-populations of poly(methyl methacrylate) (PMMA) and silica beads. The study included 20 participating laboratories from industry, academia, and government, and a variety of state-of-the-art particle-counting instruments. The received datasets were organized by instrument class to enable comparison of intralaboratory and interlaboratory performance. The main findings included high variability between datasets from different laboratories, with coefficients of variation from 13 % to 189 %. Intralaboratory variability was, on average, 37 % of the interlaboratory variability for an instrument class and particle sub-population. Drop-offs at either end of the size range and poor agreement on maximum counts of particle sub-populations were noted. The mean distributions from an instrument class, however, showed the size-coverage range for that class. The study shows that a polydisperse sample can be used to assess performance capabilities of an instrument set-up (including hardware, software, and user settings) and provides guidance for the development of polydisperse reference materials.

Development and Validation of Optical Methods for Zeta Potential Determination of Silica and Polystyrene Particles in Aqueous Suspensions.

Zeta potential is frequently used to examine the colloidal stability of particles and macromolecules in liquids. Recently, it has been suggested that zeta potential can also play an important role for grouping and read-across of nanoforms in a regulatory context. Although the measurement of zeta potential is well established, only little information is reported on key metrological principles such as validation and measurement uncertainties. This contribution presents the results of an in-house validation of the commonly used electrophoretic light scattering (ELS) and the relatively new particle tracking analysis (PTA) methods. The performance characteristics were assessed by analyzing silica and polystyrene reference materials. The ELS and PTA methods are robust and have particle mass working ranges of 0.003 mg/kg to 30 g/kg and 0.03 mg/kg to 1.5 mg/kg, respectively. Despite different measurement principles, both methods exhibit similar uncertainties for repeatability (2%), intermediate precision (3%) and trueness (4%). These results confirm that the developed methods can accurately measure the zeta potential of silica and polystyrene particles and can be transferred to other laboratories that analyze similar types of samples. If direct implementation is impossible, the elaborated methodologies may serve as a guide to help laboratories validating their own methods.

Validation of a Homogeneous Incremental Centrifugal Liquid Sedimentation Method for Size Analysis of Silica (Nano)particles.

Silica nanoparticles display many unique physicochemical properties that make them desirable for use in a wide variety of consumer products and composite materials. Accurately measuring the size of these nanoparticles is important for achieving the desired nanoscale functionality of the final product and for regulatory compliances. This study covers the validation of a centrifugal liquid sedimentation method for accurate measurement of the Stokes diameter of silica particles with a near-spherical shape and dimensions in the nanometer and sub-nanometer scale range. The validated method provided unbiased results in the range of 50 nm to 200 nm, with a lower limit of detection of ≤20 nm. The relative standard uncertainties for precision, quantified in terms of repeatability and day-to-day variation, ranged from 0.2% to 1.0% and from <0.1% to 0.5%, respectively. The standard uncertainty for trueness was assessed at 4.6%. Within its working range, the method was found robust with respect to the type of cuvette, light factor, operator, and for defining the meniscus of the sample suspension. Finally, a relative expanded measurement uncertainty of 10% confirmed the satisfactory performance of the method.

Validation of a particle tracking analysis method for the size determination of nano- and microparticles.

Particle tracking analysis (PTA) is an emerging technique suitable for size analysis of particles with external dimensions in the nano- and sub-micrometre scale range. Only limited attempts have so far been made to investigate and quantify the performance of the PTA method for particle size analysis. This article presents the results of a validation study during which selected colloidal silica and polystyrene latex reference materials with particle sizes in the range of 20 nm to 200 nm were analysed with NS500 and LM10-HSBF NanoSight instruments and video analysis software NTA 2.3 and NTA 3.0. Key performance characteristics such as working range, linearity, limit of detection, limit of quantification, sensitivity, robustness, precision and trueness were examined according to recommendations proposed by EURACHEM. A model for measurement uncertainty estimation following the principles described in ISO/IEC Guide 98-3 was used for quantifying random and systematic variations. For nominal 50 nm and 100 nm polystyrene and a nominal 80 nm silica reference materials, the relative expanded measurement uncertainties for the three measurands of interest, being the mode, median and arithmetic mean of the number-weighted particle size distribution, varied from about 10% to 12%. For the nominal 50 nm polystyrene material, the relative expanded uncertainty of the arithmetic mean of the particle size distributions increased up to 18% which was due to the presence of agglomerates. Data analysis was performed with software NTA 2.3 and NTA 3.0. The latter showed to be superior in terms of sensitivity and resolution.

Reference materials and representative test materials to develop nanoparticle characterization methods: the NanoChOp project case.

This paper describes the production and characteristics of the nanoparticle test materials prepared for common use in the collaborative research project NanoChOp (Chemical and optical characterization of nanomaterials in biological systems), in casu suspensions of silica nanoparticles and CdSe/CdS/ZnS quantum dots (QDs). This paper is the first to illustrate how to assess whether nanoparticle test materials meet the requirements of a "reference material" (ISO Guide 30, 2015) or rather those of the recently defined category of "representative test material (RTM)" (ISO/TS 16195, 2013). The NanoChOp test materials were investigated with small-angle X-ray scattering (SAXS), dynamic light scattering (DLS), and centrifugal liquid sedimentation (CLS) to establish whether they complied with the required monomodal particle size distribution. The presence of impurities, aggregates, agglomerates, and viable microorganisms in the suspensions was investigated with DLS, CLS, optical and electron microscopy and via plating on nutrient agar. Suitability of surface functionalization was investigated with attenuated total reflection Fourier transform infrared spectrometry (ATR-FTIR) and via the capacity of the nanoparticles to be fluorescently labeled or to bind antibodies. Between-unit homogeneity and stability were investigated in terms of particle size and zeta potential. This paper shows that only based on the outcome of a detailed characterization process one can raise the status of a test material to RTM or reference material, and how this status depends on its intended use.

Polyelectrolyte-coated unilamellar nanometer-sized magnetic liposomes.

Superparamagnetic nanoparticles were encapsulated in liposomes prior to the stepwise adsorption of polyelectrolytes of opposite charges commonly known as the layer-by-layer (LbL) technique. Magnetic fields allow a fast separation of coated liposomes from unbound polyelectrolytes. The coated particles were characterized by dynamic light scattering (DLS), cryo-TEM, AFM, and zeta-potential techniques. The presence of magnetic nanoparticles and the polyelectrolyte shell opens the possibility of their magnetic manipulation and targeting by applying an external magnetic or electric field.

Multifunctionalized polymer microcapsules: novel tools for biological and pharmacological applications.

We describe recent developments with multifunctional nanoengineered polymer capsules. In addition to their obvious use as a delivery system, multifunctional nanocontainers find wide application in enzymatic catalysis, controlled release, and directed drug delivery in medicine. The multifunctionality is provided by the following components: 1) Luminescent semiconductor nanocrystals (quantum dots) that facilitate imaging and identification of different capsules, 2) superparamagnetic nanoparticles that allow manipulation of the capsules in a magnetic field, 3) surface coatings, which target the capsules to desired cells, 4) metallic nanoparticles in the capsule wall that act as an absorbing antenna for electromagnetic fields and provide heat for controlled release, and 5) enzymes and pharmaceutical agents that allow specific reactions. The unique advantage of multifunctional microcapsules in comparison to other systems is that they can be simultaneously loaded/functionalized with the above components, allowing for the combination of their properties in a single object.