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In medicinal chemistry, Pd is perhaps the most-widely utilized precious metal, as catalyst in reactions which represent key transformations toward the synthesis of new active pharmaceutical ingredients (APIs). The disadvantage of this metal-catalyzed chemistry is that expensive and toxic metal residues are invariably left bound to the desired product. Thus, stringent regulatory guidelines exist for the amount of residual Pd that a drug candidate is allowed to contain. In this work, a rapid and simple method for the determination of Pd in API samples by high-energy polarized-beam energy dispersive X-ray fluorescence spectrometry has been developed and validated according to the specification limits of current legislation (10 mg [kg.sup.-1] Pd) and the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH guidelines). Sample and calibration standards preparation includes a first step of homogenization and then, in a second step, the pressing of the powdered material into pellets without any chemical treatment. The use of several synthetic calibration standards made of cellulose to simulate the API matrix appears to be an effective means to obtain reliable calibration curves with a good spread of data points over the working range. With the use of the best measuring conditions, the limit of detection (0.11 mg [kg.sup.-1] Pd) as well as the limit of quantitation (0.37 mg [kg.sup.-1] Pd) achieved meet rigorous requirements. The repeatability of the XRF measurement appeared to be less than 2%, while the precision of the whole method was around 7%. Trueness was evaluated by analyzing spiked API samples at the level of the specification limit and calculating the recovery factor, which was better than 95%. To study the applicability of the developed methodology for the intended purpose, three batches of the studied API were analyzed for their Pd content, and the attained results were comparable to those obtained by the daily routine method (acid digestion plus atomic spectroscopy) used in most pharmaceutical laboratories.

Heterozygous de novo mutations in the Activity-Dependent Neuroprotective Homeobox (ADNP) gene underlie Helsmoortel-Van der Aa syndrome (HVDAS). Most of these mutations are situated in the last exon and we previously demonstrated escape from nonsense-mediated decay by detecting mutant ADNP mRNA in patient blood. In this study, wild-type and ADNP mutants are investigated at the protein level and therefore optimal detection of the protein is required. Detection of ADNP by means of western blotting has been ambiguous with reported antibodies resulting in non-specific bands without unique ADNP signal. Validation of an N-terminal ADNP antibody (Aviva Systems) using a blocking peptide competition assay allowed to differentiate between specific and non-specific signals in different sample materials, resulting in a unique band signal around 150 kDa for ADNP, above its theoretical molecular weight of 124 kDa. Detection with different C-terminal antibodies confirmed the signals at an observed molecular weight of 150 kDa. Our antibody panel was subsequently tested by immunoblotting, comparing parental and homozygous CRISPR/Cas9 endonuclease-mediated Adnp knockout cell lines and showed disappearance of the 150 kDa signal, indicative for intact ADNP. By means of both a GFPSpark and Flag-tag N-terminally fused to a human ADNP expression vector, we detected wild-type ADNP together with mutant forms after introduction of patient mutations in E. coli expression systems by site-directed mutagenesis. Furthermore, we were also able to visualize endogenous ADNP with our C-terminal antibody panel in heterozygous cell lines carrying ADNP patient mutations, while the truncated ADNP mutants could only be detected with epitope-tag-specific antibodies, suggesting that addition of an epitope-tag possibly helps stabilizing the protein. However, western blotting of patient-derived hiPSCs, immortalized lymphoblastoid cell lines and post-mortem patient brain material failed to detect a native mutant ADNP protein. In addition, an N-terminal immunoprecipitation-competent ADNP antibody enriched truncating mutants in overexpression lysates, whereas implementation of the same method failed to enrich a possible native mutant protein in immortalized patient-derived lymphoblastoid cell lines. This study aims to shape awareness for critical assessment of mutant ADNP protein analysis in Helsmoortel-Van der Aa syndrome., (© 2024. The Author(s).)

A novel electrothermal atomic absorption spectrometry (ETAAS) method was developed for the halogenation of refractory sample components (Er, Nd and Nb) of lithium niobate (LiNbO(3)) and bismuth tellurite (Bi(2)TeO(5)) optical single crystals to overcome memory effects and carry-over. For this purpose, the cleaning step of a regular graphite furnace heating program was replaced with a halogenation cycle. In this cycle, after the graphite tube cooled to room temperature, a 20 μL aliquot of liquid carbon tetrachloride (CCl(4)) was dispensed with a conventional autosampler into the graphite tube. The CCl(4) was partially dried at 80°C under the mini-flow (40 cm(3) min(-1)) condition of the Ar internal furnace gas (IFG), then the residue was decomposed (pyrolyzed) by fast furnace heating at 1900-2100°C under interrupted flow of the IFG. This step was followed by a clean-out stage at 2100°C under the maximum flow of the IFG. The advantage of the present method is that it does not require any alteration to the graphite furnace gas supply system in contrast to most of the formerly introduced halogenation techniques. The effectiveness of the halogenation method was verified with the determination of Er and Nd dopants in the optical crystals. In these analyses, a sensitivity decrease was observed, which was likely due to the enhanced deterioration of the graphite tube surface. Therefore, the application of mathematical correction (resloping) of the calibration was also required. The calibration curves were linear up to 1.5 and 10 μmol L(-1) for Er and Nd, respectively. Characteristic masses of 18 and 241 pg and the limit of detection (LOD) values of 0.017 and 0.27 μmol L(-1) were found for Er and Nd, respectively. These LOD data correspond to 0.68 μmol mol(-1) Er and 11 μmol mol(-1) Nd in solid bismuth tellurite samples. The analytical results were compared with those obtained by a conventional ETAAS method and validated with X-ray fluorescence spectrometry analysis., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The chemical composition of airborne particulate matter (PM) was studied at a coastal region near De Haan, Belgium, during a winter-spring and a summer campaign in 2006. The major ionic components of size-segregated PM, i.e. NH(4)(+), Na(+), K(+), Mg(2+), Ca(2+), Cl(-), NO(3)(-), and SO(4)(2-), and related gaseous pollutants (SO(2), NO(2), NH(3), HNO(2), and HNO(3)) were monitored on a daily basis. Air mass backward-trajectories aided in evaluating the origin of the diurnal pollution load. This was characterised with high levels of fine secondary inorganic aerosols (NH(4)(+), NO(3)(-), and non-sea-salt SO(4)(2-)) for continental air masses, and sea-salts as the dominant species in coarse maritime aerosols. Seasonal variations in the level of major ionic species were explained by weather conditions and the release of dimethyl sulfide from marine regions. This species was responsible for an increased sea-salt Cl(-) depletion during summer (56%), causing elevated levels of HCl. Neutralisation ratios for the coarse fraction (0.6-0.8) suggested a depleted NH(4)(+) level, while that for the fine fraction (1.1-1.3) had definitely an excess of NH(4)(+), formed by the neutralisation of HCl. The results of factor analysis and the extent of SO(2) oxidation indicated that the major ionic species originated from both local and remote sources, classifying the Belgian coastal region as a combined source-receptor area of air pollution.

Atmospheric aerosols can consist of, amongst others, compounds like NH(4)NO(3) or (NH(4))(2)SO(4). Such components can suffer radiation damage and/or evaporate during EDXRF measurements, providing errors on successively applied analysis. The aim of this work is to investigate the influence of measurements using conventional EDXRF on the volatile compounds and to compare it with the influence of polarized beam EDXRF using secondary targets (and hence indirect irradiation). The effect of different parameters (acquisition time, accelerating voltage, current and medium) on the concentration loss was studied. The measurements performed in vacuum during a long period lead to the highest losses of volatile compounds. The influence of direct irradiation was proved to be larger than the indirect variant.

Total suspended particulate (TSP), PM(2.5) and BTEX were collected in nine offices in the province of Antwerp, Belgium. Both indoor and outdoor aerosol samples were analysed for their weight, elemental composition, and water-soluble fraction. Indoor TSP and PM(2.5) concentrations ranged from 7-31 microg m(-3) and 5-28 microg m(-3), with an average of 18 and 11 microg m(-3), respectively. Of all the elements analysed in indoor TSP, more than 95% was represented by Al, Si, K, Ca, Fe, Cl and S, accounting for 12% of the TSP by mass. The other elements showed significant enrichment relative to the earth's crust. The water-soluble ionic fraction accounted for almost 30% of the sampled indoor TSP by weight, and was enriched by anthropogenic activities. It was shown that the indoor PM levels varied among the offices, depending on the ventilation pattern, location, and occupation density of the office. Indoor BTEX levels ranged together from 5-47 microg m(-3) and were considerably higher than the corresponding outdoor levels. It was observed that some recently constructed and renovated buildings were clearly burdened with elevated levels for toluene, ethyl benzene, and xylenes, while outdoor air was found to be the main source for BTEX levels at the 'older' offices.

In this paper, we describe a new method for trace level Cd determination in complex environmental liquid samples. Thin layers activated with the extractant Aliquat 336 were prepared either by direct impregnation of commercial polymeric supports or by physical inclusion in a cellulose triacetate matrix, and both were effectively used to collect Cd present at low concentration in different aqueous matrixes. Quantitation of Cd contained in the thin layers was performed by high-energy polarized-beam energy-dispersive X-ray fluorescence. The effects of various experimental parameters such as layer composition, equilibration time, and instrumental conditions have been investigated. The analysis of different impregnated layers contacted with solutions ranging from 5 to 8000 microg L-1 Cd showed a linear response between the Cd concentration in the aqueous solutions and the metal present in the thin layer, with a detection limit of 0.7 microg L-1. The accuracy of the proposed method was confirmed by analyzing spiked seawater samples and a synthetic water sample containing, besides Cd, high amounts of other metal pollutants such as Ni, Cu, and Pb. The attained results were comparable to those obtained by anodic stripping voltammetry or inductively coupled plasma spectrometry.

A fast and direct determination procedure for precious metals in spent automotive catalyst was developed using the novel high-energy polarized-beam XRF. A sample preparation method working directly on the ground material was optimized. The material was pressed as a pellet using wax as a binder; no internal standard was added. The standards for this application were available spent automotive catalyst, previously analyzed by ICP-OES to verify their concentration, prepared in the same way as the unknown samples. The investigated concentration ranged from nearly 0 to approximately 2700 ppm for Pt, to 500 ppm for Rh, and to 7500 ppm for Pd. The repeatability of the XRF measurement appeared to be better than 0.5%, while the precision of the whole method was approximately 1%. The accuracy of the XRF method was verified with the well-established (but very time-consuming) ICP-OES method; a good agreement (no difference when using the 95% confidence interval) was found for the results. When using an irradiation time of 500 s for the CsI secondary target and the Zr secondary target, the detection limits for Pt, Pd, and Rh were found to be better than 5 ppm.