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Production of toxaphene enantiomers by enantioselective HPLC after isolation of the compounds from an anaerobically degraded technical mixture.

Enantiomers of 12 chlorobornanes were separated on a chiral stationary HPLC phase. The investigated compounds included relevant chlorobornanes in technical toxaphene (Toxicant A and an unknown hepatachlorobornane), anaerobically mediated media such as sediment, soil, and sewage sludge (B6-923, B7-1001), as well as eight persistent compounds of technical toxaphene (CTTs) frequently detected in biological samples (B7-1000, B7-1453, B8-1412, B8-1413 or P-26, B8-1414 or P-40, B8-1945 or P-41, B8-2229 or P-44, and B9-1679 or P-50). Sufficient amounts of these 12 CTTs were not commercially available and had to be produced in our lab. Eight CTTs were obtained from sewage sludge that was spiked with technical toxaphene and kept under anaerobic conditions for four weeks. The samples were extracted with hexane followed by RP-HPLC fractionation. The resulting toxaphene pattern was significantly simpler than that of the technical mixture. CTTs that showed intense fragmentation in GC/ECNI-MS were preferably metabolized. Moreover, only one of the diastereomers that make Toxicant A (B8-806/B8-809 or P-42a/b) resisted degradation in sewage sludge. We found that the persistent component of Toxicant A is 2,2,5-endo,6-exo,8,9,9,10-octachlorobornane (B8-809 or P-42b). B9-1679 (P-50), B7-1453, and B8-1412 were earlier isolated from biological samples, and B7-1000 was isolated from naturally contaminated sediments. The fractions obtained after these procedures were suitable for enantioselective HPLC separations. The first eluting enantiomer was usually obtained as an enantiopure standard whereas the second eluting enantiomer also contained the other enantiomer. Attempts to determine the optical rotation with the help of a chiral HPLC detector failed. Elution orders of the enantiomers were established on three GC chiral stationary phases. Only the enantiomers of B7-1453 and B8-1945 (P-41) eluted in the same order from all CSPs while the others showed different enantiomer elution orders or were not resolved on one of the chiral GC stationary phases. The knowledge and consideration of these results is important for the interpretation of enantiomer ratios found in biological samples and comparison of literature data.

Camphanes↗

Two-dimensional fluorometry coupled with artificial neural networks: a novel method for on-line monitoring of complex biological processes.

The use of two-dimensional scanning fluorometry as an on-line, noninvasive, in situ bioreactor monitoring technique is extended to complex bioprocesses using mixed cultures, with particular attention to biofilm systems. Using the example of spectra subtraction, it is demonstrated that established methods for fluorescence data analysis have a limited capability of utilizing overall fluorometric information. Artificial neural networks (ANNs) are introduced as a novel nonlinear and nonmechanistic technique for interpreting the highly complex fluorescence maps. It is shown that ANNs are able to infer process performance parameters in a pattern recognition approach, based on the entire fluorescence "fingerprint" of the biological system. The studies were carried out using an extractive membrane bioreactor (EMB) for the degradation of chlorinated organic compounds, operating with mixed cultures. Model pollutants em- ployed were 1,2-dichloroethane, 3-chloro-4-methylaniline, and p-toluidine.

Biodegradation, Environmental↗

Data mining by total ranking methods: a case study on optimisation of the "pulp and bleaching" process in the paper industry.

Total order ranking methods are multicriteria decision making techniques used for the ranking of various alternatives on the basis of more than one criterion. The criteria, which are the standards by which the elements of the system are judged are not always in agreement, they can be conflicting, motivating the need to find an overall optimum that can deviate from the optima of one or more of the single criteria. Total order ranking methods are based on an aggregation of the criteria in a scalar function, i.e. an order or ranking index, which allow to sort elements according to its numerical value. Several evaluation methods which define a ranking parameter generating a total order ranking have been proposed in the literature. Four total order ranking methods are here described: Desirability functions, Utility functions, Dominance functions and Absolute Reference method. These methods have been compared to each other by applying them to a decision making problem in the paper industry. Various bleaching processes have been analysed and compared on the basis of multiple criteria, the aim being to find out best bleaching process among the ones proposed in the last years as alternative to chlorine bleaching process which is of high environmental impact due to the potential for chlorinated dioxin production.

Case-Control Studies↗

First-aid reports of acute chlorine gassing among pulpmill workers as predictors of lung health consequences.

Workers in pulpmills can be exposed to a multitude of gases hazardous to respiratory function, the most common of which is chlorine gas. First-aid reports of acute gas overexposure incidents ("gassings") over an 8 year period were used to generate exposure data on a group of pulpmill workers whose respiratory function had been studied cross-sectionally in 1981 and 1988. Three hundred forty-eight incidents representing 174 workers were identified, 78% of these being treated solely by the first-aid attendant with the administration of O2 and cough suppression medication. Among 316 workers tested during a 1988 respiratory health survey, 78 had at least one chlorine or chlorine dioxide "gassing" incident. There was a significant decrease in the FEV1/FVC ratio (p less than .05) as well as increased risk for workplace associated chest symptoms in this group with at least one "gassing" incident. In an age- and smoking-matched analysis, among workers tested both in 1981 and 1988, there was a greater decline in FEV2/FVC ratio and MMF (p less than .05) in the "gassed" group than in the nonexposed group over the 7 year period of observation. These results emphasize the need for worker protection against accidental chlorine gas exposures.

Analysis of Variance↗

Genotoxicity of surface water treated with different disinfectants using in situ plant tests.

Disinfection of surface drinking water, in particular water chlorination, results in many by-products with potential genotoxic and/or carcinogenic activity. In the present study, we evaluated the genotoxicity of surface water after treatment with different disinfectants by means of in situ plant genotoxicity assays (micronucleus and chromosomal aberration tests) which can detect both clastogenic and aneugenic effects. The study was carried out at a pilot plant using lake water after sedimentation and filtration. This water supplied four stainless steel basins: three basins were disinfected with sodium hypochlorite, chlorine dioxide, and peracetic acid and the fourth basin containing untreated lake water was used as a control. Plants were exposed in situ in the basins. The study was carried out using water collected in different seasons over a period of about 1 year in order to assess the treatments in different physical and chemical lake water conditions. The micronucleus test in root cells of Vicia faba (Vicia faba/MCN test) revealed genotoxicity in many samples of disinfected water. The micronucleus test in Tradescantia pollen cells and the chromosome aberration test in root cells of Allium cepa showed genotoxic effects only in some disinfected samples, but also revealed genotoxicity in raw water. The results of the study indicated that the Vicia faba/MCN test was the most sensitive plant assay for disinfected water and that peracetic acid disinfection produced similar or lower genotoxicity than sodium hypochlorite or chlorine dioxide treatment.

Chlorine Compounds↗

Effect of chlorine dioxide and its metabolites in drinking water on fetal development in rats.

The chlorination of surface waters is known to form trihalomethanes. Therefore, chlorine dioxide (C1O2) is being considered as an alternative disinfectant. This study was designed to determine the effect of chlorine dioxide and its metabolites, chlorite (C1O-2) and chlorate (C1O-3), on rat fetuses exposed in utero. Female rats were administered C1O2 at 0, 1, 10 and 100 mg 1(-1) and C1O-2 or C1O-3 at 1 and 10 mg 1(-1) daily in the drinking water for 2 1/2 months prior to the throughout gestation. Rats were killed on day 20 and fetuses examined for external, skeletal and visceral malformations. Slight decreases in weight gain during pregnancy were seen in the C1O2 administered groups. A significant dose-response relationship in the decreases of the numbers of implants and live fetuses were observed in the C1O2 groups. Although there were increased incidences of resorptions in the C1O-2 and C1O-3 groups, no statistically significant increase was found in the groups. Fetal weight was significantly increased in the 100 mg 1(-1) C1O2 group. Also, fetal length was increased in the 10 mg 1(-1) C1O-2 and C1O-3 treatment groups. Skeletal defects, such as incompletely ossified or missing sternebrae, rudimentary ribs and incompletely ossified skull bones were increased in all treatment groups, but none were significantly different from the control group. A few cases of hypoplastic kidney, hydronephrosis and dextrocardia were observed in the treatment groups.

Animals↗

Effects of Alcide gel on fetal development in rats and mice. II.

Alcide gel is a germicidal agent which is highly effective in killing a wide range of bacteria and fungi in vitro within 1 min. Alcide gel consists of sodium chlorite and lactic acid, parts A and B, respectively. Chlorine dioxide is formed when sodium chlorite and lactic acid, the active ingredients of Alcide, are mixed. The objective of this investigation was to study the teratogenic effects of Alcide gel both in rats and in mice. The gel was administered topically in doses of 1 and 2 g per kg body weight on days 6-15 of gestation. No toxic symptoms were observed following exposure of pregnant rats and mice to Alcide gel. Viability of the fetuses as well as weight, length and number of resorptions were quite similar compared with control groups. Fetuses were examined for external, visceral and skeletal anomalies. Some incidence of missing or incompletely ossified sternebrae, missing ribs and incomplete ossification of skull bones were observed from skeletal examination. Gastromegaly, kidney agenesis, heart displacement, interatrial septal defect, limb hyperflexion and club foot were observed from soft-tissue examination. These defects, however, were not significantly different compared with control animals. Alcide gel was not teratogenic at the doses tested.

Abnormalities, Drug-Induced↗

The effect of Alcide, a new antimicrobial drug, on rat blood glutathione and erythrocyte osmotic fragility, in vitro.

Alcide is an antimicrobial drug which has been demonstrated to kill a variety of common pathogenic bacteria as well as fungi, in vitro. This agent is supplied in liquid and gel forms and consists of two parts, one of which contains sodium chlorite, while the other contains lactic acid as the active ingredients. Mixing of the two parts prior to use produces chlorine dioxide (ClO2), a strong oxidizing agent. A dose-dependent decrease in glutathione content and erythrocyte osmotic fragility occurred after incubation of whole blood with Alcide. Glutathione concentration and erythrocyte osmotic fragility approached the control values after 240 min of incubation with Alcide containing 1 mM NaClO2. The addition of exogenous glutathione (50 mg 100 ml-1) or glutathione reductase and NADPH to rat blood in the presence of Alcide returned erythrocyte osmotic fragility to control values. Treatment of rat blood with Alcide did not change glutathione reductase or glutathione peroxidase activities after 1 h of incubation.

Animals↗

Subchronic dermal toxicity studies of Alcide Allay gel and liquid in rabbits.

Alcide is a germicidal agent which is highly effective in killing a wide range of bacteria and fungi. 2.0 g kg-1 Allay gel or placebo were applied once per day while 3.0 g kg-1 Allay liquid or placebo were administered three times per day over a 30 day period to the skin of albino rabbits. Allay gel and liquid contained either of two concentrations of sodium chlorite and lactic acid as active ingredients. The concentration of active ingredients, sodium chlorite and lactic acid, in the low dose gel was 25% of high dose gel and low dose liquid was 60% of high dose liquid. Moderate to severe erythema was observed only in the high dose gel group after 7 days of treatment, but skin appeared visibly normal by day 18. Histologically fixed skin at day 30, however, showed inflammatory changes in the high and low dose gel groups and hyperkeratosis in all gel groups. At the termination of the study, mean corpuscular hemoglobin concentration decreased significantly in the Allay gel and liquid groups compared to an untreated control group. In all gel treatments, BUN/creatinine levels decreased significantly. Hematology and clinical chemistry parameters, however, were within the normal range of values for the gel and liquid groups, indicating no clinically significant changes due to Allay treatment. Pancreas/body weight ratios were significantly reduced in all gel groups, while spleen, pancreas and ovary/body weight ratios were significantly higher in both liquid dosage groups. No histological changes were observed in any of these organs.

Animals↗

Biaxial flex-fatigue and viral penetration of natural rubber latex gloves before and after artificial aging.

Barrier integrity of unaged and oven-aged (at 70 degrees C) natural rubber latex examination gloves was assessed with a biaxial flex-fatigue method where failure was detected electronically, and by live viral penetration testing performed according to a modified version of ASTM F1671-97a. When no change in barrier properties was detected during flex testing, no virus passage was found after viral challenge. Conversely, when a change in the barrier properties was indicated by the electrical signal, virus passage was found in 74% of the specimens. Flex-fatigue results indicated that unaged test specimens from powdered (PD) and powder-free (PF) nonchlorinated gloves had significantly longer fatigue lives than powder-free chlorinated (CL) gloves from the same manufacturer. Biaxial flexing of oven-aged glove specimens showed a marginal increase in fatigue life for the PF gloves, but no increase for the PD gloves. The fatigue life of the CL gloves was observed to increase significantly after oven aging. However, this appears to be due to a design feature of the test apparatus, wherein peak volume displacement of the worked specimen is held constant. An aging-induced change in the viscoelastic properties of the CL gloves-permanent deformation of the specimens early in the fatigue test-relieves the stress magnitude applied as the test progresses. Thus, permanent deformation acts as a confounding factor in measuring durability of latex gloves by fixed displacement flex-fatigue.

Bacteriophage phi X 174↗

Sepsis of vascular catheters. II: In vitro disinfection of colonized tubing.

Preparatory to development of in situ disinfection of implanted catheters, silicone rubber tubing colonized by incubation with Staphylococcus aureus, Staphylococcus epidermidis, or Klebsiella pneumoniae was used to test the efficacy of various chemicals in vitro. Protocols sterilizing all segments colonized for 24 h (n = 30) were immersion into 50% povidone iodine for 5 and 60 min, 100% povidone iodine for 5, 15, and 60 min, 1.2 x 10(3) ppm chlorine dioxide for 15 and 60 min, and 1.2 x 10(3) ppm chlorine dioxide buffered to pH 5.1 for 60 min. Immersion in up to 2% chlorhexidine, 7.4% formaldehyde, or 6% sodium hypochlorite for up to 60 min failed to sterilize all segments. None of 117 control segments were sterilized. Segments colonized for seven days were sterilized by immersion into 100% povidone iodine for 15 or 60 min. Use of 1.2 x 10(3) ppm chlorine dioxide for 60 min sterilized 97% of segments tested. Lower concentrations and shorter exposure times failed to sterilize all segments. Eighteen silicone rubber catheters, colonized on the outer surface, were all sterilized within 24 h by a chlorine dioxide solution placed in the lumen and diffusing through the wall to kill the bacteria.

Catheterization↗

Analysis of dichloroacetic acid in rat blood and tissues by hydrophilic interaction liquid chromatography with tandem mass spectrometry.

Dichloroacetic acid (DCA) is a compound found in chlorinated drinking water. In addition, the compound is a metabolite of several halogenated solvents, including trichloroethylene (TCE) and perchloroethylene (PCE). Exposure to DCA is of concern because high doses of the compound have been shown to cause cancer in laboratory animals. Dosages of TCE administered to animals in cancer studies are designed to elicit maximal DCA formation in vivo, whereas levels of DCA to which individuals are exposed in drinking water are very low. Analysis of DCA in biological samples has been quite challenging. Derivatizing reagents commonly used to convert DCA into a more volatile form for analysis by gas chromatography (GC) have been found to convert trichloroacetic acid (TCA), a major metabolite of TCE and PCE, into DCA. High-performance liquid chromatography (HPLC) analysis does not require derivatization of DCA and can thus avoid this problem. However, the most popular stationary phases in HPLC columns do not retain small, polar compounds such as DCA well. The liquid chromatography/tandem mass spectrometry (LC/MS/MS) method described in this paper uses hydrophilic interaction liquid chromatography (HILIC), a type of chromatography that is able to retain these small, polar compounds. Method validation was performed using the United States Food and Drug Administration (USFDA) and International Conference on Harmonziation (ICH) Guidance for Industry: Bioanalytical Method Validation as a guide. Levels of DCA found in rats dosed with 2 g/kg TCE were 17.2 ng/mL (liver), 262.4 ng/mL (kidney), 175.1 ng/mL (lung), and 39.5 ng/mL (blood).

Animals↗

Influence of drinking water composition on quantitation and biological activity of dissolved microcystin (cyanotoxin).

Toxic cyanobacteria in aquatic environments have been implicated in many poisoning incidents of livestock, wildlife, and domestic animals. Microcystins (MCYSTs) in water supplies represent a risk to public health. This work investigated the effect of water composition on the quantitation and biological activity of MCYSTs analyzed by different methods (HPLC, ELISA, and protein phosphatase 1 inhibition assay). Different MCYST concentrations were added to deionized water and quantified, confirming the efficiency of these analytical methods. MCYST concentrations diluted in drinking water had reduced detection by all methods tested. The drinking water used contained a free chlorine concentration of 2.5 mg/L and an Fe concentration of 0.45 mg/L, and the conductivity was 69.8 microS cm(-1), whereas in deionized water, free chlorine and Fe were not detectable, and the conductivity was 1.6 microS cm(-1). Drinking water also interfered with the biological activity of MYCSTs, as these toxins showed reduced protein phosphatase-1 inhibition. A free chlorine concentration of 2.5 mg/L in deionized water was completely effective in preventing any detection of 10 microg/L of added MCYSTs. Fe and Al ions also were very effective in reducing MCYST detection. The chemical composition of drinking water thus affected MCYST detection, indicating a significant reduction in quantitation of this molecule either because of its decomposition or through complexation with metal ions.

Chlorine Compounds↗

Application of diluted chlorine dioxide to radish and lettuce nurseries insignificantly reduced plant development.

The possible toxicity of a commercial chlorine dioxide preparation (Halox E-100) was evaluated on radish and lettuce seedlings growing in pots under controlled conditions. A single application of various dilutions to radish seedlings growing in a sterile or nonsterile commercial plant substrate only slightly decreased plant dry weight. At the end of the experiments, the plants appeared unaffected by the treatments. Other common plant parameters (root and stem length, number of true leaves) were unaffected or even enhanced. Halox did not reduce the total level of soil bacteria even after four consecutive applications at any dilution rate. In nonsterile soil, high Halox dilution (1:1000) significantly decreased plant dry weight, and the other concentrations (1:10,000; 1:50,000, and 1:100,000) had no apparent effect on the size of the plants. In sterile soil, high concentrations of Halox (1:1000 and 1:10,000) significantly decreased plant growth, but higher dilutions produced no significant reduction in plant dry weight. For radish plants growing in organic matter-free sand only, dilution of 1:10,000 reduced plant growth. On lettuce plants, dilutions from 1:5000 to 1:25,000 did not reduce plant growth. High levels of Halox (1:1000) were toxic to both radish and lettuce seedlings growing in sand and resulted in chlorosis and significant depression of plant growth. Further dilutions of Halox (equivalent to the level used in water disinfection) significantly decreased toxicity for both plant species. Low concentrations of Halox (> 1:50,000) had no apparent effect on the appearance of both plant species. In conclusion, this study suggests that chlorine dioxide-treated drinking water can be considered safe for growing plants; this treatment should be further evaluated using other plant species under more realistic growth conditions.

Chlorine↗

Dietary exposure to chemical contaminants from traditional food among adult Dene/Métis in the western Northwest Territories, Canada.

Environmental contaminants such as organochlorines and heavy metals have been reported to bioaccumulate in Arctic and subarctic wildlife. The Indigenous Peoples in northern and Arctic Canada rely on local wildlife as an important food source, and it is thus hypothesized that they may have high intakes of these contaminants. Herein, an assessment of dietary exposure to selected organochlorines and heavy metals for Indigenous Peoples of the western Northwest Territories (NWT) is presented. Dietary data were collected from 1012 adults with 24-h recalls in 16 communities in the western NWT (Denendeh). A comprehensive survey of the literature, as well as in-house analysis, formed the basis of a large traditional food-contaminant database. By combining the dietary and contaminant data, dietary exposure to 11 chemical contaminants was calculated. Dietary exposure to chemical contaminants in Denendeh is generally low and there is little, if any, associated health risk. However there are specific contaminants in certain communities for which exposure on a single day approaches the tolerable daily intake levels. These situations are detailed and monitoring needs are described.

Adult↗