Paving the way for new and challenging matrix reference materials-particle suspensions at the core of material processing providing RMs for method development and method validation.

Sufficient homogeneity of the certified parameter(s) over the whole fill series of a matrix reference material (RM) is a fundamental quality criterion. In practice, the heterogeneity of the target parameter is evaluated, whereby a relative value can be calculated of how much the target parameter is varying over the RM-batch. A high degree of homogeneity (low heterogeneity) is an inherent quality mark of a good RM. Here, we report how challenging matrix RMs were produced by using particle suspensions at the core of the material processing step. The examples of matrix RMs produced span from whole water reference materials for persistent organic pollutants, PM2.5-like atmospheric dust certified for specific ions to microplastic RMs. Most of these RMs were subsequently used in different phases of analytical method development or for method validation. Common to all these matrices is that they cannot be easily mixed, handled, or dosed to prepare larger sample batches. In all cases, a continuously stirred suspension of particles was used during material processing. In general, relative between-bottle heterogeneities from 1.6 to 6% were achieved for the target parameters in these matrix presentations. Concerning developments of new CRMs in emerging fields, the co-dependence between the availability of validated analytical methods with good repeatability and testing materials with a known and high homogeneity of the target parameter(s) becomes particularly challenging. This situation is an RM/Method causality dilemma. To overcome that hurdle, strategies are proposed for stepwise processes where RM producers and a network of analytical method developers could work hand in hand. In addition, development of a portfolio of inexpensive and well-homogenised common samples coupled with a reporting interface is suggested. This would benefit method developers and RM producers alike. As more and more data is compiled for a specific matrix, it paves the way for new and challenging RMs that can later be used by a wider community.

Infrared Thermography for Monitoring of Freeze Drying Processes-Part 2: Monitoring of Temperature on the Surface and Vertically in Cuvettes during Freeze Drying of a Pharmaceutical Formulation.

The coupling of an infrared (IR) camera to a freeze dryer for monitoring of the temperature of a pharmaceutical formulation (sucrose/mannitol solution, 4:1%, m/m) during freeze-drying has been exploited further. The new development allows monitoring of temperatures simultaneously at the surface as well as vertically, (e.g., in depth) along the side using custom-made cuvettes. The IR camera was placed on the chamber roof of a process-scale freeze dryer. Monitoring of cuvettes containing the formulation took place from above where one side of each cuvette was equipped with a germanium window. The Ge-window was placed next to an IR mirror having a 45° angle. The long-wave infrared radiation (LWIR) coming from the inside of the cuvette was reflected upwards toward the IR camera. Accurate recording of the temperature along the cuvettes' depth profile was therefore possible. Direct imaging from -40 °C to 30 °C took place every 60 s on the surface and on the side with a 2 × 2 mm resolution per IR pixel for 45 h resulting in 2700 thermograms. Results are presented for freeze-drying of a pharmaceutical formulation as a function of time and spatially for the entire side (depth) of the cuvette. As the sublimation process was progressing, the spatial resolution (84 IR pixels for the side-view and 64 pixels for the surface-view) was more than sufficient to reveal lower temperatures deeper down in the material. The results show that the pharmaceutical formulation (a true solution at the onset) dries irregularly and that the sublimation front does not progress evenly through the material. During secondary drying, potential evaporative cooling of upper layers could be detected thanks to the high thermal and spatial resolution.

First amyloid ß1-42 certified reference material for re-calibrating commercial immunoassays.

INTRODUCTION: Reference materials based on human cerebrospinal fluid were certified for the mass concentration of amyloid beta (Aß)1-42 (Aß42 ). They are intended to be used to calibrate diagnostic assays for Aß42 . METHODS: The three certified reference materials (CRMs), ERM-DA480/IFCC, ERM-DA481/IFCC and ERM-DA482/IFCC, were prepared at three concentration levels and characterized using isotope dilution mass spectrometry methods. Roche, EUROIMMUN, and Fujirebio used the three CRMs to re-calibrate their immunoassays. RESULTS: The certified Aß42 mass concentrations in ERM-DA480/IFCC, ERM-DA481/IFCC, and ERM-DA482/IFCC are 0.45, 0.72, and 1.22 µg/L, respectively, with expanded uncertainties (k = 2) of 0.07, 0.11, and 0.18 µg/L, respectively. Before re-calibration, a good correlation (Pearson's r > 0.97), yet large biases, were observed between results from different commercial assays. After re-calibration the between-assay bias was reduced to < 5%. DISCUSSION: The Aß42 CRMs can ensure the equivalence of results between methods and across platforms for the measurement of Aß42 .

Preparation of a PM2.5-like reference material in sufficient quantities for accurate monitoring of anions and cations in fine atmospheric dust.

A reference material of a PM2.5-like atmospheric dust material has been prepared using a newly developed method. It is intended to certify values for the mass fraction of SO42-, NO3-, Cl- (anions) and Na+, K+, NH4+, Ca2+, Mg2+ (cations) in this material. A successful route for the preparation of the candidate reference material is described alongside with two alternative approaches that were abandoned. First, a PM10-like suspension was allowed to stand for 72 h. Next, 90% of the volume was siphoned off. The suspension was spiked with appropriate levels of the desired ions just prior to drop-wise shock-freezing in liquid nitrogen. Finally, freeze drying of the resulting ice kernels took place. In using this approach, it was possible to produce about 500 g of PM2.5-like material with appropriate characteristics. Fine dust in 150-mg portions was filled into vials under an inert atmosphere. The final candidate material approaches the EN12341 standard of a PM2.5-material containing the ions mentioned in Directive 2008/50/EC of the European Union. The material should be analysed using the CEN/TR 16269:2011 method for anions and cations in PM2.5 collected on filters. The method described here is a relatively rapid means to obtain large quantities of PM2.5. With access to smaller freeze dryers, still 5 to 10 g per freeze-drying cycle can be obtained. Access to such quantities of PM2.5-like material could potentially be used for different kinds of experiments when performing research in this field. Graphical abstract The novelty of the method lies in transformation of a suspension with fine particulate matter to a homogeneous and stable powder with characteristics similar to air-sampled PM2,5. The high material yield in a relatively short time is a distinct advantage in comparison with collection of air-sampled PM2,5.

Certified reference materials for testing of the presence/absence of Staphylococcus aureus enterotoxin A (SEA) in cheese.

Staphylococcal enterotoxins (SEs) account for a substantial number of food-poisoning outbreaks. European legislation (Commission Regulation 1441/2007) stipulates the reference procedure for SE analysis in milk and dairy products, which is based on extraction, dialysis concentration and immunochemical detection using one of two approved assays (VIDAS(®) SET2, Ridascreen(®) SET Total). However, certified reference materials (CRMs) are lacking to support laboratories in performing reliable detection of Staphylococcus aureus enterotoxin A (SEA) in relevant matrices at sub-nanogram per gram levels. The certification of a set of three reference materials (blank and two SEA-containing materials) for testing of the presence/absence of SEA in cheese is described. The reference procedure was applied in an intercomparison with 15 laboratories, and results were reported in a qualitative manner (presence or absence of SEA in the sample). No false-negative or false-positive results were obtained. The certified values were stated as diagnostic specificity (blank material) or diagnostic sensitivity (SEA-containing materials) and were 100 % in all cases. Stability studies demonstrated suitable material stability when stored cooled or frozen. An in-house study on the recovery of SEA in the cheese materials using a double-sandwich enzyme-linked immunosorbent assay (ELISA) revealed comparable recovery values of around 45 % at the two spiking levels and in both the SEA-containing CRMs as well as blank CRM freshly spiked prior to analysis. The values were also comparable over time and among different analysts. The materials provide valuable support to laboratories for method validation and method performance verification and will increase the reliability of measuring SEA in cheese.

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.

Infrared thermography for monitoring of freeze-drying processes: instrumental developments and preliminary results.

Coupling an infrared (IR) camera to a freeze dryer for on-line monitoring of freeze-drying cycles is described for the first time. Normally, product temperature is measured using a few invasive Pt-100 probes, resulting in poor spatial resolution. To overcome this, an IR camera was placed on a process-scale freeze dryer. Imaging took place every 120 s through a Germanium window comprising 30,000 measurement points obtained contact-free from -40 °C to 25 °C. Results are presented for an empty system, bulk drying of cheese slurry, and drying of 1 mL human serum in 150 vials. During freezing of the empty system, differences of more than 5 °C were measured on the shelf. Adding a tray to the empty system, a difference of more than 8 °C was observed. These temperature differences probably cause different ice structures affecting the drying speed during sublimation. A temperature difference of maximum 13 °C was observed in bulk mode during sublimation. When drying in vials, differences of more than 10 °C were observed. Gradually, the large temperature differences disappeared during secondary drying and products were transformed into uniformly dry cakes. The experimental data show that the IR camera is a highly versatile on-line monitoring tool for different kinds of freeze-drying processes.

Production of reference materials for the detection and size determination of silica nanoparticles in tomato soup.

A set of four reference materials for the detection and quantification of silica nanoparticles (NPs) in food was produced as a proof of principle exercise. Neat silica suspensions were ampouled, tested for homogeneity and stability, and characterized for total silica content as well as particle diameter by dynamic light scattering (DLS), electron microscopy (EM), gas-phase electrophoretic molecular mobility analysis (GEMMA), and field-flow fractionation coupled with an inductively coupled mass spectrometer (FFF-ICPMS). Tomato soup was prepared from ingredients free of engineered nanoparticles and was spiked at two concentration levels with the silica NP suspension. Homogeneity of these materials was found sufficient to act as reference materials and the materials are sufficiently stable to allow long-term storage and distribution at ambient temperature, providing proof of principle of the feasibility of producing liquid food reference materials for the detection of nanoparticles. The spiked soups were characterized for particle diameter by EM and FFF-ICPMS (one material only), as well as for the total silica content. Although questions regarding the trueness of the results from EM and FFF-ICPMS procedures remain, the data obtained indicate that even assigning values should eventually be feasible. The materials can therefore be regarded as the first step towards certified reference materials for silica nanoparticles in a food matrix.

Feed premix: a difficult matrix for the accurate determination of trace elements - the outcome of IMEP-114 and IMEP-36.

This paper presents the outcome of two proficiency tests (IMEP-114/36) running in parallel, which focused on the determination of total As, Cd, Pb, Hg and Sn in feed premixes. Both exercises aimed to test the performance of laboratories measuring trace elements in feed, in compliance with Directive 2002/32/EC of the European Parliament and of the Council on undesirable substances in animal feed. IMEP-114 was run for the European Union National Reference Laboratories, while IMEP-36 was open to all food control laboratories that wished to participate. In total 80 laboratories from 37 countries registered to both proficiency tests, from which 75 laboratories reported results. The test material used in this exercise was commercially available feed premix, which was processed, bottled, labelled and dispatched to participants. The performance of the participating laboratories was similar in both proficiency tests and was very good in the case of total Cd and less satisfactory for total As and total Pb. For total Sn only one-third of the participants reported results, from which 33% performed satisfactorily. Thirty-four participants reported results for total Hg, although the expert laboratories stated that the mass fraction for that measurand was below the limit of detection of the method used. An evaluation of the reasons that could be at the basis of unsatisfactory results has been carried out and is presented here.

Determination of dissolved bromate in drinking water by ion chromatography and post column reaction: interlaboratory study.

A collaborative study, International Evaluation Measurement Programme-25a, was conducted in accordance with international protocols to determine the performance characteristics of an analytical method for the determination of dissolved bromate in drinking water. The method should fulfill the analytical requirements of Council Directive 98/83/EC (referred to in this work as the Drinking Water Directive; DWD). The new draft standard method under investigation is based on ion chromatography followed by post-column reaction and UV detection. The collaborating laboratories used the Draft International Organization for Standardization (ISO)/Draft International Standard (DIS) 11206 document. The existing standard method (ISO 15061:2001) is based on ion chromatography using suppressed conductivity detection, in which a preconcentration step may be required for the determination of bromate concentrations as low as 3 to 5 microg/L. The new method includes a dilution step that reduces the matrix effects, thus allowing the determination of bromate concentrations down to 0.5 microg/L. Furthermore, the method aims to minimize any potential interference of chlorite ions. The collaborative study investigated different types of drinking water, such as soft, hard, and mineral water. Other types of water, such as raw water (untreated), swimming pool water, a blank (named river water), and a bromate standard solution, were included as test samples. All test matrixes except the swimming pool water were spiked with high-purity potassium bromate to obtain bromate concentrations ranging from 1.67 to 10.0 microg/L. Swimming pool water was not spiked, as this water was incurred with bromate. Test samples were dispatched to 17 laboratories from nine different countries. Sixteen participants reported results. The repeatability RSD (RSD(r)) ranged from 1.2 to 4.1%, while the reproducibility RSD (RSDR) ranged from 2.3 to 5.9%. These precision characteristics compare favorably with those of ISO 15601. A thorough comparison of the performance characteristics is presented in this report. All method performance characteristics obtained in the frame of this collaborative study indicate that the draft ISO/DIS 11206 standard method meets the requirements set down by the DWD. It can, therefore, be considered to fit its intended analytical purpose.