Method for the determination of five toxicologically relevant arsenic species in human urine by liquid chromatography-hydride generation atomic absorption spectrometry.

An analytical method for the simultaneous quantitation of arseneous acid (As(III)), arsenic acid (As(V)), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA) and trimethylarsine oxide (TMAO) in human urine by coupling of high-performance liquid chromatography with hydride generation atomic absorption spectrometry (HPLC/HG-AAS) via a flow-injection interface is presented. After arsenic species separation by anion-exchange displacement chromatography the compounds are on-line reduced to their corresponding hydrides and detected by atomic absorption spectrometry. Detection limits range from 1.1 (TMAO) to 2.6 microg/L (As(V)). The method has been applied to determine arsenic species in the urine of a volunteer before and after consumption of seafood as well as to analyse certified reference urine samples for their arsenic species content.

Determination of dichloroanilines in human urine by GC-MS, GC-MS-MS, and GC-ECD as markers of low-level pesticide exposure.

Methods for the determination of 3,4-dichloroaniline (3,4-DCA) and 3,5-dichloroaniline (3,5-DCA) as common markers of eight non-persistent pesticides in human urine are presented. 3,5-DCA is a marker for the exposure to the fungicides vinclozolin, procymidone, iprodione, and chlozolinate. Furthermore the herbicides diuron, linuron, neburon, and propanil are covered using their common marker 3,4-DCA. The urine samples were treated by basic hydrolysis to degrade all pesticides, metabolites, and their conjugates containing the intact moieties completely to the corresponding dichloroanilines. After addition of the internal standard 4-chloro-2-methylaniline, simultaneous steam distillation extraction (SDE) followed by liquid-liquid extraction (LLE) was carried out to produce, concentrate and purify the dichloroaniline moieties. Gas chromatography (GC) with mass spectrometric (MS) and tandem mass spectrometric (MS-MS) detection and also detection with an electron-capture detector (ECD) after derivatisation with heptafluorobutyric anhydride (HFBA) were employed for separation, detection, and identification. Limit of detection of the GC-MS-MS and the GC-ECD methods was 0.03 and 0.05 microg/l, respectively. Absolute recoveries obtained from a urine sample spiked with the internal standard, 3,5-, and 3,4-DCA, ranged from 93 to 103% with 9-18% coefficient of variation. The three detection techniques were compared concerning their performance, expenditure and suitability for their application in human biomonitoring studies. The described procedure has been successfully applied for the determination of 3,4- and 3,5-DCA in the urine of nonoccupationally exposed volunteers. The 3,4-DCA levels in these urine samples ranged between 0.13 and 0.34 microg/g creatinine or 0.11 and 0.56 microg/l, while those for 3,5-DCA were between 0.39 and 3.33 microg/g creatinine or 0.17 and 1.17 microg/l.

Reduced levels of 1,25-dihydroxyvitamin D(3) in rat dams and offspring after exposure to a reconstituted PCB mixture.

Previous studies revealed effects of polychlorinated biphenyls (PCBs) and other polyhalogenated hydrocarbons on steroid hormone levels and hormone-dependent functions including behavior. In the present study serum concentrations of the vitamin D(3) metabolites 25-hydroxycholecalciferol (25-D) and 1,25-dihydroxycholecalciferol (1,25-D) were determined in rat dams and offspring after exposure to a PCB mixture that was reconstituted according to the congener pattern found in human breast milk. Unmated females were exposed to diets adulterated with 0; 5; 20; or 40 mg PCBs/kg diet. Exposure started 50 days prior to mating and was terminated at birth. Gestational exposure reduced serum concentrations of 1,25-D in dams in a dose-dependent manner. Concentration of 25-D was also decreased at the time of delivery, but not at weaning. Determination of 1,25-D in offspring at weaning revealed reductions in both high-exposure groups. Levels of 25-D were diminished only at the highest exposure level. Internal PCB concentrations in adipose tissue and brains exhibited a linear relation to dosages in diet. Concentrations of PCBs in brains were similar in dams and offspring at birth, but decreased at the end of lactation in dams. In offspring, values increased during this period because of continued exposure via the milk. In the adipose tissue, PCB levels were much lower in offspring than in dams. To our knowledge, this is the first report of PCB-induced effects on vitamin D(3) metabolites. In dams, reductions were seen even at the lowest exposure level used. Further studies are needed to evaluate the biological significance of these reductions in pregnant dams and possible consequences for the developing offspring.

Ambient Particulate Matter Induces Oxidative Dna Damage in Lung Epithelial Cells.

Although epidemiological studies have established a correlation between PMIO levels and acute cardiovascular and respiratory complications, hardly any data is available on possible chronic effects such as cancer. The purpose of this study was to investigate the production of free radicals by ambient particulate matter (TSP) and to link these data to oxidative DNA damage in lung epithelial cells. In line with previous findings on PMIO, supercoiled plasmid DNA was depleted by JSP as well as JSP supernatant (p < .001), and this effect was reduced in the presence of mannitol (5 mM). Using electron spin resonance (ESR) and the spin trap dimethyl-1-pyrroline N-oxide (DMPO) we were able to show that hydroxy/radicals ('OH) are formed from both JSP and JSP supernatant. The DMPO-OH signal was completely abrogated when TSP was preincubated with deferoxamine (5 mM), showing the importance of iron and other soluble metals in this process. Atomic absorption spectroscopy (AAS) analysis of the TSP supernatant showed the presence of soluble Fe, V, and Ni (respectively 253.0, 14.7, and 76.0 µ/g insoluble TSP). To investigate the biological significance of OH formation by TSP, 8-hydroxydeoxyguanosine (8-oxodC) was measured in a rat type II cell line by immunocytochemistry. The formation of this hydroxyl-radical-specific DNA adduct was increased twofold (p < .01) after incubation with TSP supernatants, and this effect was inhibited by deferoxamine (p < .01). In summary, our results provide direct evidence that ambient particulate matter generates hydroxyI radicals in acellular systems. Furthermore, we showed that these particulates induce the hydroxyl-radical-specific DNA lesion 8-oxodC in lung target cells via an iron-mediated mechanism.

Internal platinum, palladium, and gold exposure in environmentally and occupationally exposed persons.

In a pilot study the urinary platinum (Pt), palladium (Pd), and gold (Au) excretion was analyzed in 27 dental technicians, 17 road construction workers and 17 school-leavers using sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). Detection limits in urine were 0.24 ng/l for Pt and Au and 0.17 ng/l regarding Pd. A standardized questionnaire was used to assess information about kind and degree of contact to these metals, the physical condition of the volunteers and confounding factors. Significant differences between the three study groups were found. The mean Pt, Pd, and Au excretions of the dental technicians were significantly higher than those of the road construction workers and school-leavers. This indicates that the occupational treatment of dental alloys leads to an internal exposure to these metals which is distinctly higher than that from automobile exhaust exposure. Significant differences between Monday morning (pre-shift) and Thursday afternoon (post-shift) urine samples of the dental technicians were not found. The Pt excretion of road construction workers working near a much traveled highway was comparable with that of school-leavers being less (only environmentally) exposed to automobile exhaust. Regarding Pd and Au the road construction workers showed a tendency to higher levels in urine when compared with the school-leavers, but statistically significant differences were not found. The tendency to higher urinary Pd and Au levels in the road-construction workers may be explained by their slightly greater number of noble metal containing artificial dentures, which may cause an additional exposure. A statistically significant effect of age on the urinary noble metal excretion was not detectable.

Developmental exposure of rats to a reconstituted PCB mixture or aroclor 1254: effects on organ weights, aromatase activity, sex hormone levels, and sweet preference behavior.

Polychlorinated biphenyls (PCBs) are lipophilic industrial chemicals which are regularly detected in human breast milk, serum, and tissues. They possess hormone-modulating properties, and, when transferred transplacentally to the developing fetus, PCBs have been shown to induce persistent sex-specific neurobehavioral deficits. Interactions of PCBs with sex steroid-modulated neural differentiation could in part account for such effects. To test this hypothesis, female Long-Evans rats were exposed via food containing 40 mg/kg of either a reconstituted PCB mixture (RM), composed according to the congener-pattern in human breast milk, or the technical PCB mixture Aroclor 1254 (A1254). The exposure period started 50 days prior to mating and was terminated at birth (postnatal day 0: PND 0). Aromatase (CYP 19) activity was determined in hypothalamus/preoptic area (HPOA) brain-sections from newborn male pups. This enzyme converts testosterone (T) to 17beta-estradiol (E(2)) and plays a key role in sexual brain differentiation. Moreover, serum concentrations of T and E(2), physical development, organ weights, exposure levels, and sex-specific behavior were evaluated at different life stages. On PND 0, a reduced aromatase activity was detected in the HPOA of male RM-pups compared to controls. Female RM-weanlings exhibited significantly elevated uterine wet weights on PND 21, which is a marker for estrogenic activity. In the adult stage (PND 170), male offspring with maternal exposure to either PCB mixture showed markedly reduced testes weights and serum testosterone levels, thus demonstrating persistent antiandrogenic effects. On PND 180, male RM-rats exhibited a behavioral feminization in a sweet preference test, suggesting long-lasting changes in neuronal brain organization caused by the perinatally suppressed aromatase activity. The results suggest that maternal exposure to the RM, the pattern of which is similar to the PCB spectrum in human milk, results in more distinct effects on sex steroid-dependent processes and behavior than the technical PCB mixture A1254. PCB levels in brain and adipose tissue of the exposed offspring lay within 1-2 orders of magnitude above background concentrations in humans.

Determination of selected microbial volatile organic compounds by diffusive sampling and dual-column capillary GC-FID--a new feasible approach for the detection of an exposure to indoor mould fungi?

A new, analytically valid procedure is described to assess the exposure of human beings to the so-called microbial volatile organic compounds (MVOCs) in air. The method can be used routinely for large sample numbers and is especially valuable as a basis for further research on the correlation between single MVOCs and indoor mould growth. The procedure is based on the fact that fungi produce a variety of volatile organic compounds, such as 3-methylbutan-1-ol, 3-methylbutan-2-ol, fenchone, heptan-2-one, hexan-2-one, octan-3-one, octan-3-ol, pentan-2-ol, alpha-terpineol, and thujopsene, which they emit into the indoor environment. Using diffusive samplers, these MVOCs are adsorbed onto charcoal during a sampling interval of four weeks. The described method is thus superior to existing methods which use short-term active sampling. After desorption with carbon disulfide, the MVOCs were determined by dual-column gas chromatography with flame ionization detection using the large-volume injection technique for sample introduction. The detection limits ranged between 0.15 and 0.53 microgram m-3, within-series precision was found to range between 6.5 and 19.0%, and recovery was between 77 and 118%. The procedure has been successfully applied in the context of a large field study to measure the indoor MVOC exposure in children's rooms of 132 dwellings. The objective of the study was to examine the relation between indoor mould growth, the indoor MVOC exposure and the prevalence of adverse health effects. Information about mould formation has been obtained by a questionnaire and by the determination of colony forming units of mould fungi in mattress dust. With the exception of 3-methylbutan-2-ol, fenchone, nonan-2-one, octan-2-one, and thujopsene, indoor air concentrations of all MVOCs under investigation were significantly higher inside damp and mouldy dwellings. From the primary MVOCs under investigation, 3-methylbutan-1-ol, hexan-2-one, heptan-2-one, and octan-3-ol were found to be most reliable indicators for mould formation. A correlation was also found between selected MVOCs and the occurrence of mould species in mattress dust. Aspergillus sp. correlated with heptan-2-one, hexan-2-one, octan-3-ol, octan-3-one, and alpha-terpineol, while the occurrence of Eurotium sp. was correlated with higher indoor air concentrations of 3-methylbutan-1-ol, 3-methylbutan-2-ol, heptan-2-one, hexan-2-one, octan-3-ol, and thujopsene. Children living in dwellings with elevated MVOC levels had a higher prevalence of asthma, hay fever, wheezing, and irritations of the eyes. These positive associations persisted after controlling for confounding factors such as age, sex, body-mass index, number of siblings, social status, passive smoking, type of heating, and ventilation habits. However, they were not statistically significant. This lack of significance may be a result of the small number of investigated samples.

Determination of physiological levels of volatile organic compounds in blood using static headspace capillary gas chromatography with serial triple detection.

A static capillary gas chromatographic method using three different detectors [photoionization detector (PID), electron capture detector (ECD) and flame ionization detector (FID)] switched in series is presented for the determination of volatile organic compounds (VOCs) in sub microgram l-1 levels. The method was applied for the analysis of selected environmentally and occupationally relevant non-halogenated and chlorinated aromatic hydrocarbons (e.g., benzene, toluene, xylenes, dichlorobenzenes) as well as chlorinated aliphatic hydrocarbons (e.g., trichloroethene, tetrachloroethene) in blood samples. Detailed investigations, in respect to the figures of merit were carried out. For most of the selected VOCs detection limits (calculated as the three-fold standard deviation of low level calibration standards) in the range from 26 (benzene) to 67 ng l-1 (m/p-xylene) were achieved which are comparable with those reported for dynamic headspace techniques in combination with mass spectrometric detection. For the individual VOCs the within-series precision varied from 4 to 19% and the day-to-day precision from 11 to 28%. Regarding PID as well as FID the calibration graphs for all substances were linear up to at least 10 micrograms l-1 while the ECD response was linear up to concentrations of about 0.6 microgram l-1 for the halogenated compounds. Our method is applicable for the quantitative determination of VOCs in blood in the occupationally as well as in the physiologically relevant (normal) concentration range.

Determination of benzene, toluene, ethylbenzene and xylenes in indoor air at environmental levels using diffusive samplers in combination with headspace solid-phase microextraction and high-resolution gas chromatography-flame ionization detection.

An improved analytical method for passive air sampling is presented based on a combination of commercially available diffusive samplers with headspace solid-phase microextraction and high-resolution gas chromatography with flame ionization detection (HRGC-FID). This procedure is targeted for short-term BTEX (benzene, toluene, ethylbenzene and o-, m- and p-xylenes) determinations at environmental concentrations and can be applied for sampling intervals between 30 min and 24 h. The analytes are adsorbed onto the charcoal pad of a passive sampler and then extracted with carbon disulphide-methanol. After removal of the carbon disulphide by xanthation, the BTEXs are enriched on a Carboxen SPME fiber, thermally desorbed and analysed by HRGC-FID. Detection limits for a sampling interval of 2 h are between 0.4 and 2 micrograms/m3, within-series precision ranges between 6.6 and 12.8%, day-to-day precision is between 11.1 and 15.2%. The results obtained with this procedure are validated by comparison with active sampling. Detection limits and a further reduction of the sampling time are limited by blanks of the chemicals and the diffusive samplers. Procedures to eliminate these blanks are described in detail. Applications such as the determination of BTEXs in indoor air inside buildings, inside a train and a car are presented, indicating the usefulness of the described procedure for short-term measurements of environmental BTEX concentrations. An advantage of passive samplers is the storage stability for at least six months, which is essential for its use in large epidemiological studies.

Strain differences in tissue concentrations of mercury in inbred mice treated with mercuric chloride.

Tissue concentrations of mercury were determined by cold vapor atomic absorption spectrometry in different inbred mouse strains after continuous treatment with HgCl2 (3 weekly sc injections of 0.5 mg/kg bw) for up to 12 weeks. Except for the thymus, in which steadily increasing mercury concentrations were found, in steady state levels of mercury were reached in blood and liver after 4 weeks and in spleen and kidney after 8 weeks. In the closely related strains C57BL/6, B10.D2, and B10.S, which differ only or primarily at the major histocompatibility complex, mercury concentrations in blood and liver were about twofold lower and renal concentrations were about three- to fivefold lower than those detected in strains A.SW and DBA/2. Another strain difference was observed in the spleen: after 8 and 12 weeks of continuous HgCl2 treatment, mercury concentrations in the spleen of strains A.SW, C57BL/6, and B10.S were significantly higher than those in strains DBA/2 and B10.D2. The strain difference in the spleen, an organ of the immune system, correlates with the susceptibility to the HgCl2-induced systemic autoimmune syndrome in mice in that the strains showing a higher mercury accumulation in the spleen are susceptible to this form of chemically induced autoimmunity, whereas the strains with lower mercury concentrations in the spleen are resistant.

Sensitive gas chromatographic method for determination of mercapturic acids in human urine.

A method was developed for sensitive determination of the specific benzene metabolite S-phenylmercapturic acid and the corresponding toluene metabolite S-benzylmercapturic acid in human urine for non-occupational and occupational exposure. The sample preparation procedure consists of liquid extraction of urine samples followed by precolumn derivatization and a clean-up by normal-phase HPLC. Determination of analytes occurs by gas chromatography with electron-capture detection. With this highly sensitive method (detection limits 60 and 65 ng/l, respectively) urinary S-phenylmercapturic and S-benzylmercapturic acid concentrations for non-occupationally exposed persons (e.g. non-smokers) can be measured precisely in one chromatographic run. Validation of the method occurred by comparison with a HPLC method we have published recently.

Determination of polychlorinated biphenyls and chlorinated pesticides in human body fluids and tissues.

A fast and reliable method for the determination of polychlorinated biphenyls (PCBs) and chlorinated pesticides in human body fluids and tissues is presented. Sample clean-up and selective enrichment of analytes are carried out by liquid column chromatography without a prior solvent extraction step. Analytes are determined either by high-resolution capillary gas chromatography with mass spectrometric detection, or by dual-column GC followed by electron-capture detection. A procedure is described for the simultaneous determination of environmental levels of the PCB congeners 28, 52, 101, 138, 153 and 180 in human cord serum. The method allows the simultaneous determination of chlorinated pesticides such as aldrin, DDT, DDE, BHC and HCB and is also applicable to other biological matrices, such as bone marrow or tissues.

[Gas chromatography and mass spectrometry method for detection of selected pyrethroid metabolites in urine]. / Gaschromatographische und massenspektrometrische Methode zum Nachweis ausgewählter Pyrethroidmetabolite im Urin.

Pyrethroids are increasingly used indoors, because of their low mammalian toxicity but high insecticidal activity. Indoor application of pyrethroids, which is often carried out without adequate expert knowledge, may lead to high pesticide residues, often combined with health hazards for the people concerned. Objective of the present study is the development of a method for a biological monitoring to determine internal exposure to pyrethroids. Major pyrethroid metabolites were detected in urine by capillary gas chromatography in combination with mass selective detection (MSD). For the metabolites, the limits of determination range between 0.5 and 1 microgram/L urine. Our results so far demonstrate the detectability of cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid (DVCA) and 3-phenoxybenzoic acid (3-PBA) in urine during a period of elimination of 48 hours after exposure to cypermethrin. In this paper the method is exemplary illustrated by one exposed person (pest control operator).

Internal and external tetrachloroethene exposure of persons living in differently polluted areas of Northrhine-Westphalia (Germany).

An epidemiological study was performed to measure the internal and external tetrachloroethene exposure of persons living in two differently polluted areas of Northrhine-Westphalia (Germany). Tetrachloroethene concentrations were determined in venous blood samples of 5- to 7-year-old children (n = 81) and 55-year-old women (n = 91) living in Essen, an industrial city located in the Ruhr area. 103 children und 131 women of the same age living in Borken, a small town north of the Ruhr area, served as reference group. Outdoor air samples were collected on passive samplers (sampling period: 4 weeks) from 70 measurement points per study area (about 2 km2, mean distance 100 m). In the course of a year these measurements were repeated three times to cover seasonal variations. Parallel to the outdoor measurement periods, indoor air concentrations were determined in the homes of those women from Essen and Borken, who donored a blood sample. Tetrachloroethene levels in blood were generally low with a geometric mean of 0.05 microgram/L in women and 0.021 microgram/L in children. Nevertheless, children and women living in the industrial area were found to have significantly higher tetrachloroethene levels in blood than those of the reference group. In both study areas blood levels of women exceeded those of children by a factor of 2. Participants living in the neighbourhood of a dry-cleaning shop had distinctly elevated blood levels. The same applied to persons who stored dry-cleaned clothes at home. Like the internal exposure, external exposure was also higher in Essen than in Borken. In both areas tetrachloroethene concentrations indoors exceeded those outdoors. Outdoor tetrachloroethence concentrations were significantly increased during the cold season, while the opposite was true for indoor levels. The correlation between indoor and outdoor exposure was found to be significant, while those between blood levels and outdoor exposure became only significant when people living next to a dry-cleaning shop were excluded. No significant relationship was observed between blood and indoor tetrachlorethene levels. It is concluded that the higher tetrachloroethene blood levels of the urban population result from the higher atmospheric concentrations in industrial areas with tetrachloroethene emitting sources like metal and textile industry. The fact that indoor air tetrachloroethene levels exceeded those outdoors can only be explained by the presence of additional indoor sources. Provided that women spend on average more time indoors than children the higher indoor air concentrations may be the reason for the higher blood tetrachloroethene levels found in women. Persons living near a dry-cleaning shop or storing dry-cleaned clothes at home showed a higher internal and external exposure to tetrachloroethene than other persons. In individual cases it can by far exceed the average exposure of the general population, so that health impairments can not be generally excluded.

Long-term mercury excretion in urine after removal of amalgam fillings.

The long-term urinary mercury excretion was determined in 17 28- to 55-year-old persons before and at varying times (up to 14 months) after removal of all (4-24) dental amalgam fillings. Before removal the urinary mercury excretion correlated with the number of amalgam fillings. In the immediate post-removal phase (up to 6 days after removal) a mean increase of 30% was observed. Within 12 months the geometric mean of the mercury excretion was reduced by a factor of 5 from 1.44 micrograms/g (range: 0.57-4.38 micrograms/g) to 0.36 microgram/g (range: 0.13-0.88 microgram/g). After cessation of exposure to dental amalgam the mean half-life was 95 days. These results show that the release of mercury from dental amalgam contributes predominantly to the mercury exposure of non-occupationally exposed persons. The exposure from amalgam fillings thus exceeds the exposure from food, air and beverages. Within 12 months after removal of all amalgam fillings the participants showed substantially lower urinary mercury levels which were comparable to those found in subjects who have never had dental amalgam fillings. A relationship between the urinary mercury excretion and adverse effects was not found. Differences in the frequency of effects between the pre- and the post-removal phase were not observed.

Internal lead and cadmium exposure in 6-year-old children from western and eastern Germany.

Lead and cadmium levels in blood and deciduous teeth (shed incisors only) of 6-year-old German children were determined in 1991 in a large epidemiological study carried out in rural and urban areas of western Germany (Duisburg, Essen, Gelsenkirchen, Dortmund, Borken) and eastern Germany (Leipzig, Halle, Magdeburg, Osterburg, Gardelelegen, Salzwedel). In total, blood lead and cadmium levels of 2311 German children and tooth lead and cadmium levels of 790 German children were analyzed. Blood lead levels were generally low in all study areas with geometric means between 39.3 micrograms/l and 50.8 micrograms/l in the western German and between 42.3 micrograms/l and 68.1 micrograms/l in the eastern German study areas. The mean blood lead level of Turkish children (n = 213) living in the western German study areas was 50.1 micrograms/l and thus 5.6 micrograms/l higher than the overall geometric mean of the western German children. The higher exposure may be explained by a higher oral uptake from food and different living conditions. These children were excluded from multiple regression analysis because they were all living in the western study areas. The mean tooth lead levels ranged between 1.50 and 1.74 micrograms/g in the western and between 1.51 micrograms/g and 2.72 micrograms/g in the eastern study areas. Thus, they show a distribution pattern similar to blood. Blood and tooth lead levels were higher in urban than in rural areas and higher in the eastern German than in the western German study areas. With regard to the blood and tooth cadmium concentrations, no significant differences between the study areas could be found.(ABSTRACT TRUNCATED AT 250 WORDS)

Redox Chemistry and [Au(CN)(2)] in the Formation of Gold Metabolites.

The role of hypochlorite ion, which can be generated by the enzyme myleoperoxidase, in the biochemistry of gold(I) anti-arthritic drugs was investigated. Sodium hypochlorite (OCl(-)) directly and rapidly oxidizes AuSTm, Au(CN)(2) (-), AuSTg (gold thioglucose) and auranofin (Et(3)PAuSATg). The resulting gold(III) species were detected by an Ion Chromotography Ion-Pairing technique that was developed to distinguish gold(I) and gold(III). Formation of Au(III) was also demonstrated spectrophotometrically after the conversion to AuCl(4) (-). The reactions of AuSTm, AuSTg, and auranofin are complex and gold(III) appears only after the initial oxidation of the thiolate (and phosphine) ligands.The enzymatic reaction, using MPO with H(2)O(2) and Cl(-) as substrates, leads to slow oxidation of Au(CN)(2) (-), AuSTm or AuSTg. The extent and rate of reaction depend on the concentrations of MPO, H(2)O(2), and Au(I). The continued presence of Au(I) during the initial stages of reaction (oxidation of the thiolates in AuSTm and AuSTg) and the conversion to Au(III) in the latter stages of the reaction were demonstrated. Au(CN)(2) (-), a gold metabolite, binds tightly to serum albumin. Unlike other gold(I) complexes, aurocyanide reacts almost negligibly at Cys-34 via ligand exchange. Instead, there is a strong association (K(1) = 5.5 x 10(4) and K(2) = 7.0 x 10(3); n(1) = 0.8 and n(2) = 3) of intact Au(CN)(2) (-). The full extent of binding is revealed only by equilibrium methods such as NMR or ultrafiltration; the bound gold dissociates extensively on conventional gel-exclusion columns and partially on Penefesky spun columns.The immunological and pharmacological significance of these results are discussed.