Separation and enrichment of palladium and gold in biological and environmental samples, adapted to the determination by total reflection X-ray fluorescence.

The reductive co-precipitation of trace and ultra-trace elements together with mercury followed by complete evaporation of the mercury makes it possible to determine palladium and gold by total reflection X-ray fluorescence. Both elements can be detected without interferences at optimal sensitivity in the pg range. Thus, detection limits of, e.g., 2.5 ng L-1 for palladium and 2.0 ng L-1 for gold, in urine, were obtained. The precision was determined to 0.04 at a palladium concentration of about 200 ng L-1 urine and to 0.19 at a gold concentration of only 18 ng L-1. The recovery for a urine sample spiked with known amounts of palladium and gold amounted to > 95%. Results of the combined procedure are given for the determination of palladium and gold in the urine of non-exposed and occupationally exposed persons and in some other environmentally relevant samples.

Characterization of a new type of sulfite dehydrogenase from Paracoccus pantotrophus GB17.

The periplasmic sulfite dehydrogenase of Paracoccus pantotrophus GB17 was purified to homogeneity by a four-step procedure from cells grown lithoautotrophically with thiosulfate. The molecular mass of native sulfite dehydrogenase was 190 kDa as determined by native gradient PAGE. SDS-PAGE showed sulfite dehydrogenase to comprise two subunits with molecular masses of 47 kDa and 50 kDa, suggesting an alpha2beta2 structure. The N-terminal amino acid sequence and immunochemical analysis using SoxC-specific antibodies identified the 47-kDa protein as the soxC gene product. SoxD-specific antibodies identified the 50-kDa protein as SoxD. Based on the molecular masses deduced from the nucleotide sequence for mature SoxC (43,442 Da) and SoxD (37,637 Da) sulfite dehydrogenase contained 1.30 mol molybdenum/mol alpha2beta2 sulfite dehydrogenase. The iron content was 3.17 mol/mol alpha2beta2 sulfite dehydrogenase, and 3.53 mol heme/mol alpha2beta2 sulfite dehydrogenase was determined by pyridine hemochrome analysis. These data are consistent with the two heme-binding domains (CxxCH), characteristic for c-type cytochromes, deduced from the soxD nucleotide sequence. Electrospray ionization revealed two masses for SoxC of 43,503 and 43,897 Da. The difference in molecular mass was attributed to the molybdenum cofactor of SoxC. For SoxD a mass of 38,815 Da was determined; this accounted for the polypeptide and two covalently bound hemes. Reconstitution of the catalytic activity of sulfite dehydrogenase required additional fractions; these eluted from Q Sepharose at 0.05, 0.25, and 0.30 M NaCl. The K(m) of sulfite dehydrogenase for sulfite was 7.0 microM and for cytochrome c 19 microM. Sulfite dehydrogenase activity was inhibited by sulfate and phosphate. The structural and catalytic properties make sulfite dehydrogenase from P. denitrificans GB17 distinct from sulfite oxidases of other prokaryotic or eukaryotic sources.

Ultramicroanalysis of dental plaque films by total reflection X-ray fluorescence.

Microgram quantities of dental plaque were taken near amalgam fillings, gold crowns and intact teeth. Such extremely small samples can be analysed by total reflection X-ray fluorescence (TXRF), a fairly new variant of energy dispersive X-ray fluorescence (EDXRF). More than sixty samples were examined directly without chemical pretreatment. Fifteen elements of interest were detected simultaneously within a wide range of mass fraction and with detection limits of several mg/kg. A significant correlation of the Hg-accumulation in plaque and the amalgam fillings was established. Near these fillings Hg mass fractions can reach a level of 300 mg/kg. The results for other elements, e.g. Au, are less significant.

Determination of trace elements in blood serum of patients with Behçet disease by total reflection x-ray fluorescence analysis.

The distribution of trace elements Fe, Cu, Zn, Se, and Rb was determined in sera from patients with the complete type of Behçet disease in the active stage and from healthy controls. Total reflection x-ray fluorescence was used for quantitative analysis; in general, atomic absorption spectrometry was used only for control of the Se values. Zn, Se, and Rb concentrations were significantly (P < 0.0001) lower for patients than for control subjects. No significant difference was determined in the Fe content. However, there was a significant increase in Cu in the patients so that the ratios Cu/Zn, Cu/Se, and Cu/Rb were significantly higher for the patients than for the control subjects (P < 0.0001).

Quantification of biomineralization: an in-vitro tissue culture system and microanalysis of calcium, phosphorus and trace elements by total-reflection X-ray fluorescence.

An in-vitro tissue culture system with folded periostea of 17-day-old fetal chick calvaria was combined with analytical methods to achieve quantification of biomineralization. A scanning electron microscope with an energy-dispersive X-ray detector was applied to show the distribution of calcium, phosphorus and trace elements. Calcium and phosphorus were concentrated in the zone of the mineralized matrix. Strontium was distributed similar to calcium. Zinc was distributed equally in the soft tissue and the mineralized matrix. Total-reflection X-ray fluorescence was used for quantification. Thyroxine in high concentration reduces the calcium content of the samples. One week after incubation magnesiumchloride (1.8 mM) or zincchloride (.1 mM) were found to reduce the calcium content by 38% or 82%, respectively.

Qualitative survey analysis of thin layers of tissue samples. Heavy metal traces in human lung tissue.

A special X-ray fluorescence spectrometric method has been used for the analyses of thin layers of tissue samples. This method [called Total-Reflection X-ray Fluorescence (TXRF)] gives qualitative information about numerous trace elements of physiological and environmental interest. Microtome cuts of frozen lung tissue samples are placed on quartz glass targets and excited to X-ray fluorescence by the primary beam of a molybdenum tube. This method is of interest due to the very short time needed for sampling, sample preparation and simultaneous measurement. It could be a helpful procedure in pathological and occupational medicine if a decision has to be made as to whether or not a tissue sample is extremely dust loaden.