Dioxin procedure

An environmental protocol has been developed to assess the significance of newly discovered hazardous substances that might enter soil, water, and the food chain. Using established laboratory procedures and C-labeled 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD), gas chromatography, and mass spectrometry, we determined mobility of TCDD by soil TLC in five soils, rate and amount of plant uptake in oats and soybeans, photodecomposition rate and nature of the products, persistence in two soils at 1,10, and 100 ppm, and metabolism rate in soils. We found that TCDD is immobile in soils, not readily taken up by plants, subject to photodecomposition, persistent in soils, and slowly degraded in soils to polar metabolites. Subsequent studies revealed that the environmental contamination by TCDD is extremely small and not detectable in biological samples. 

The most convenient and successful synthetic preparation of octa-chlorodibenzo-p-dioxin has been described by Kulka (13). The procedure involves chlorination of pentachlorophenol in refluxing trichlorobenzene to give octachlorodibenzo-p-dioxin in 80% yield. Kulka has explained the reaction as coupling between two pentachlorophenoxy radicals. Large amounts (5—15%) of heptachlorodibenzo-p-dioxin were observed in the unpurified product. Since the pentachlorophenol used in this study contained 0.07% tetrachlorophenol, we feel that tetrachloro-phenol may be produced in situ (Reaction 4). Such a scheme would be analogous to the formation of 2,4-dichlorophenol and 3-chlorophenol produced from 2,4,4 -trichloro-2 -hydroxydiphenyl ether (Reaction 2). The solubility of octachlorodibenzo-p-dioxin was determined in various solvents data are presented in Table II. 

Since 1950 many in the chemical industry have been keenly aware of the possibilities of highly toxic 2,3,7,8-tetrachlorodibenzo-p-dioxin forming in the manufacture of 2,4,5-trichlorophenol. This phenol is made by treating 1,2,4,5-tetrachlorobenzene with strong caustic at high temperatures for several hours. Manufacturing procedures which do not carefully control temperature and alkalinity increase the probability of dioxin formation. 

Miriyala and Williamson have described the synthesis of /i-kctocarboxam idcs from primary and secondary amines and 2,2-dimethyl-2H,4H-l,3-dioxin-4-ones as reactive a-oxoketene precursors (Scheme 6.158) [304], The experimental procedure involved heating a mixture of the dioxinone with 2-3 equivalents of the amine at ca. 180 °C for 1-3 min under solvent-free conditions in a sealed vessel by microwave irradiation. A small collection of 18 /3-ketocarboxamides was prepared in very high yields using this protocol. 

Also the production of specific Abs for PCDDs/PCDFs has been directed toward the development of immunoaffinity procedures [248,249]. Shelver et al. reported several works regarding the use of IAC to selectively extract and analyze these compounds from complex matrices such as milk or serum [250-253]. Moreover, a separation of very similar dioxin congeners (i.e., 1,3,7,8-TCDD and 2,3,7,8-TCDD) was also examined [254]. 

Two-dimensional GC can be used to separate complex mixtures of polyaromatic compounds, and MS used to subsequently identify the compounds. In this method, the original sample is injected into a gas chromatograph with one type of column. As the components exit the first GC, they are fed into a second GC, with a different column, for further separation and finally into a mass spectrometer. In this way, compounds that coeluted from the first column are separated on the second. Focant et al. [19] were able to separate polychlorinated dibenzo-p-dioxin (PCDD), polychlorinated dibenzofuran (PCDF), and coplanar polychlorinated biphenyl (cPCB) using this type of analytical procedure, including isotope dilution TOF-MS. These compounds are frequently found as contaminants in soils surrounding industrial settings thus, the ability to separate and identify them is extremely important [6,12,19],... 

Korfmacher et al. [71] employed a short clean-up procedure followed by electron capture gas chromatography for the determination of octachlorodibenzo-p-dioxin in soils using a furzed silica capillary gas chromatographic column. The technique was suitable as a routine screening procedure for samples taken from contaminated sites. 

Christman et al. [72] gave details of procedures for extraction, clean-up, and concentration of samples of soil prior to the determination of their content of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans by gas chromatography and by gas chromatography-mass spectrometry. Instrumental parameters are also included. Some typical results are tabulated. 

The benzene-water extraction gas chromatographic procedure described in section 2.1.1.1 for the determination of aliphatic hydrocarbons in soil has also been applied to the determination of polychlorinated dibenzo-p-dioxins in soil [73],... 

Representative multiple ion mass chromatograms of soil samples are presented in Fig. 5.4. These gas chromatography-mass spectrometric determinations of polychlorodibenzo-p-dioxin and polychlorodibenzofurans, and non-ortho polychlorobiphenyls in differing types of samples serve to exemplify the versatility of the procedure for such analyses. The gas chromatography-mass spectrometric data were usually uncluttered by extraneous components, and interpretation of the data was routinely straightforward. 

Volume 75 concludes with six procedures for the preparation of valuable building blocks. The first, 6,7-DIHYDROCYCLOPENTA-l,3-DIOXIN-5(4H)-ONE, serves as an effective /3-keto vinyl cation equivalent when subjected to reductive and alkylative 1,3-carbonyl transpositions. 3-CYCLOPENTENE-l-CARBOXYLIC ACID, the second procedure in this series, is prepared via the reaction of dimethyl malonate and cis-l,4-dichloro-2-butene, followed by hydrolysis and decarboxylation. The use of tetrahaloarenes as diaryne equivalents for the potential construction of molecular belts, collars, and strips is demonstrated with the preparation of anti- and syn-l,4,5,8-TETRAHYDROANTHRACENE 1,4 5,8-DIEPOXIDES. Also of potential interest to the organic materials community is 8,8-DICYANOHEPTAFULVENE, prepared by the condensation of cycloheptatrienylium tetrafluoroborate with bromomalononitrile. The preparation of 2-PHENYL-l-PYRROLINE, an important heterocycle for the synthesis of a variety of alkaloids and pyrroloisoquinoline antidepressants, illustrates the utility of the inexpensive N-vinylpyrrolidin-2-one as an effective 3-aminopropyl carbanion equivalent. The final preparation in Volume 75, cis-4a(S), 8a(R)-PERHYDRO-6(2H)-ISOQUINOLINONES, il lustrates the conversion of quinine via oxidative degradation to meroquinene esters that are subsequently cyclized to N-acylated cis-perhydroisoquinolones and as such represent attractive building blocks now readily available in the pool of chiral substrates. 

DPE suits, and butyl rubber (NRC, 2001b). The levels were 0.0002 to 0.0008 ng/m3, three orders of magnitude below the EPA criterion of 0.2 ng/m3 for dioxin emissions from incinerators. In the test with neat GB, the product gas contained 0.01 to 0.06 percent phosphine. As noted previously in the section on methods development testing, phosphine can interfere with the measurement of GB. Based on results from the EPA s toxicity characteristic leachate procedure, stabilization would be necessary only for solid wastes derived from DPE suit material, because the cadmium and lead criteria were not met by the treated dunnage in some tests (NRC, 2001b). 

Specific bans on chemicals or uses have not been the most important outcome of TSCA. Only one type of chemical, PCBs, was specifically targeted in the original law and they are now outlawed in most of their uses. EPA administration of the law in its early years led to a ban of chlorofluorocarbons as aerosol propellants, restrictions on dioxin waste disposal, rules on asbestos use, and testing rules on chlorinated solvents. It has led to a central bank of information on existing commercial chemicals, procedures for further testing of hazardous chemicals, and detailed permit requirements for submission of proposed new commercial chemicals. 

The interlaboratory results obtained from the analysis of defined standard solutions, but also from the analysis of sediment extracts prepared either by the coordinator of the study or by the participants themselves, also provide a measure of the variation between laboratories. The results show that the interlaboratory reproducibility ranges from 6.5% for the defined dioxin sample to 27.9% for the sediment sample extracted by the participants themselves. As was mentioned before, the reproducibility for this last sample is relatively high and most presumably due to the introduction of extra handlings (extraction and cleanup) to the total procedure. In addition, the fact that not all the participants had prior experience with the extraction protocol to be used could have added to the increase in variability of the process. Furthermore, the dilution factor was not dictated. This also introduces a certain degree of variation. For the reproducibility of the DR CALUX bioassay itself and not caused by differences in operating extraction conditions, the maximum variation between laboratories was observed to be 18.0%. The results for the sediment extract samples can also be used to estimate the method variability for extracts, that is, based on samples of unknown composition. Again, given the intra-as well as the interlaboratory variations observed in this study, it appears justified to conclude that the standard deviation of the means provides a reasonable estimate of the method variability, based on the overall aver-... 

The in vitro bioassay for dioxins with cleaned sediment extracts (DR-CALUX) proved to comply with the QA/QC criteria needed to guarantee the reliability of data in an inter- and intralaboratory study (Besselink et al., 2004). The chemical stability of dioxins makes it possible to apply destructive clean-up procedures which remove all matrix factors. Sample extraction and cleanup for other in vitro bioassays for specific mechanisms of toxicity require further development to make sure that the chemicals of interest are not lost or unwanted chemicals included in the sediment extract to be tested. Table 4 summarizes possible bioassays that could be performed in addition to chemical analyses with the dredged sediment in a licensing system. 

For example, in eel, high concentrations of polychlorinated biphenyls can be present and can interfere in the cleanup procedure. Fly ash is difficult to extract. Drastic concentration and cleanup procedures such as saponification can convert some isomers. Oc-tachlorodibenzo-p-dioxin can easily be broken down during the cleanup procedure. For each matrix a specific isolation technique is necessary. The method has to be validated for all the isomers of the analytical program (24). 

In 1995, the amendments to the Clean Air Act included requirements for maximum achievable control technology (MACT) for pollutants, including dioxins and metals. The EPA has since promulgated regulations that will have to be met by a modified baseline process for the Pueblo site (EPA, 1999). In addition, EPA is reviewing the effects of dioxins on human health because some evidence indicates dioxins may be more harmful in the food chain than previously thought (Kaiser, 2000). If dioxin emission standards are modified, analytical procedures will need to be reviewed. 

A publication summarises all the then available technical evidence related to the Seveso accident, and recommends operational criteria to ensure safety in commercial processes to produce trichlorophenol [4]. All the plant scale incidents were characterised [1] by the subsequent occurrence of chloracne arising from the extremely toxic and dermatitic compound 2,3,7,8-tetrachlorodibenzodioxin, formed during the thermal runaway reaction and dispersed in the ensuing explosion. It is also extremely resistant to normal chemical decontamination procedures, and after the 1968 explosion, further cases occurred after transient contact with plant which had been decontaminated and allowed to weather for 3 years, and which appeared free of dioxin [7,9]. The consequences at Seveso 447 cases of chemical burns (NaOH) and 179 of chloracne, only 34 with both [17],... 

Miellet [80] and Lopez-Avila et al. [81] have reviewed the applications of Soxhlet extraction to the determination of pesticides in soil. This technique has been applied extensively to the extraction of polycyclic aromatic hydrocarbons, volatile organic compounds, pesticides, herbicides and polychlorodibenzo-p-dioxins in soils. Details of the extraction procedures and the analytical finish employed are reviewed in Table 1.1. 

Other food samples like egg, animal fat and fish oil appear to behave very similarly to milk fat and extensive validation has not been carried out thus far. The suitability was, however, demonstrated by inclusion of positive or spiked samples. The assay was validated for blood samples from wildlife species for high concentrations of dioxins and dioxin-like PCBs.13 A special clean-up procedure was developed and validated for sediment, pore water and other environmental samples, allowing the use of the assay for official testing of these type of samples.14... 

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