Evidence for cometabolic transformation of weathered toxaphene under aerobic conditions using camphor as a co-substrate.

AIMS: Toxaphene is a persistent organic pollutant, composed of approximately 1000 highly chlorinated bicyclic terpenes. The purpose of this study was to evaluate if camphor, a structural analogue of toxaphene, could stimulate aerobic biotransformation of weathered toxaphene. METHODS AND RESULTS: Two enrichment cultures that degrade camphor as the sole carbon source were established from contaminated soil and biosolids. These cultures were used to evaluate aerobic transformation of weathered toxaphene. Only the biosolids culture could transform compounds of technical toxaphene (CTTs) in the presence of camphor, while no transformation was observed in the presence of glucose or with toxaphene as a sole carbon source. The transformed toxaphene had lower concentration of CTTs with longer retention times, and higher concentration of compounds with lower retention times. Gas chromatography with electron capture negative ion mass spectrometry (GC/ECNI-MS) showed that aerobic biotransformation mainly occurred with Cl8 - and Cl9 -CTTs compounds. The patterns of Cl6 - and Cl7 -CTTs were also simplified albeit to a much lesser extent. Seven camphor-degrading bacteria were isolated from the enrichment culture but none of them could degrade toxaphene. CONCLUSION: Camphor degrading culture can aerobically transform CCTs via reductive pathway probably by co-metabolism using camphor as a co-substrate. SIGNIFICANCE AND IMPACT OF THE STUDY: Since camphor is naturally produced by different plants, this study suggests that stimulation of aerobic transformation of toxaphene may occur in nature. Moreover plants, which produce camphor or similar compounds, might be used in bioremediation of contaminated soils.

Genotoxicity in fishes environmentally exposed to As, Se, Hg, Pb, Cr and toxaphene in the lower Colorado River basin, at Mexicali valley, Baja California, México.

The environmental exposure to As, Se, Hg, Pb, Cr and toxaphene was assessed for 11 freshwater fish species in irrigation channels, agricultural return flow drains, a drain collecting lagoon and sections of the Colorado River at the Mexicali valley in Baja California, México, during August 2015-April 2016. Arsenic (2.90 ng ml-1) and Se (1.41 ng ml-1) in water had the highest concentrations in the return flow drains (Hardy River and Xochimilco Lagoon, respectively). However, fish axial muscle tissue had the highest concentration of Se (8.3 µg g-1) and Hg (0.36 µg g-1) in Colorado River fresh water, while As (1.7 µg g-1) in Hardy River fish was highest. Selenium concentrations in all fishes and toxaphene in Cyprinus carpio and Ameiurus natalis are above the safe levels for human consumption (0.3 µg g-1 and 180 ng g-1 respectively). Toxaphene was detected in the fish axial tissue, having the highest concentrations in Poecilia latipinna (690 ng g-1) in the Colorado River. The low proportion of the 8-Cl toxaphene congeners in fish suggests degradation of this pollutant. Tilapia. sp. cf. zillii had the most genotoxic damage with 7.4 micronucleated erythrocytes per 10,000 erythrocytes in Xochimilco Lagoon and 2 in Hardy River. The genotoxicity in all the fish species studied was significantly correlated to the concentrations of As and Se in water.

Development of Biomonitoring Equivalents for chlordane and toxaphene with application to the general Canadian population.

Biomonitoring Equivalents (BEs) were developed for chlordane and toxaphene using one-compartment pharmacokinetic models and compared with biomonitoring data from the Canadian Health Measures Survey, Cycle 1 (2007-2009). A secondary objective was to examine the toxicities of the components of technical chlordane in a HEPG2 cell culture experiment. Oral reference doses were identified from national and international regulatory agencies and sources. Pharmacokinetic parameters were obtained from experimental data in rodent models. A set of BEs have been derived for the main chlordane isomers, cis-chlordane, trans-chlordane, cis-nonachlor, and trans-nonachlor, and the chlordane metabolite, oxychlordane. BEs were also derived for the main toxaphene isomers found in biota, Parlar No. 26, 50 and 62. Among the general Canadian population, no exceedances of chlordane or toxaphene BEs were observed. Based on the LC50 from the in vitro study, trans-nonachlor was the most toxic, and the trans-isomers were more toxic than the cis-isomers. The derived BE values can be used as screening guidelines to assess the risk of biomonitoring data in human populations. The results of an in vitro experiment suggest that trans-nonachlor is more toxic than technical chlordane and, therefore, the BE for this compound may need to be further lowered.

Analysis and Characterization of Polychlorinated Hydroxybornanes as Metabolites of Toxaphene Using a Polar Bear Model.

Abiotic and biotic transformation of toxaphene (camphechlor) results in the selective enrichment of recalcitrant congeners while other, less persistent compounds of technical toxaphene (CTTs) are degraded. Until now, there has been little knowledge on oxidation transformation of toxaphene. For instance, the existence of hydroxylated CTTs (OH-CTTs) in authentic environmental and food samples has not been proven. For this reason, we synthesized a mixture consisting of tetra- to heptachlorinated OH-CTTs and simplified it by countercurrent chromatography (CCC). Thus, 227 OH-CTTs were detected in the CCC fractions (12 tetra-, 117 penta-, 81 hexa-, and 17 heptachlorinated OH-CTTs), which was >50% more than detected before the fractionation. One CCC fraction consisting of only 18 OH-CTTs was used to develop a sample cleanup method which aimed to remove CTTs, isobaric PCBs, and sample matrix. The final cleanup procedure consisted of (i) gel permeation chromatography (GPC) and adsorption chromatography using (ii) deactivated and (iii) activated silica gel. Hence, up to 320 and 4350 µg/kg lipid weight of octa- and nonachlorinated CTTs were detected in four liver samples and adipose tissue of polar bears, respectively. Furthermore, the presence of one hexachlorinated OH-CTT isomer could be verified in the samples, which was about 1% of the octachlorinated CTTs determined in the liver samples.

Toxaphene-induced mouse liver tumorigenesis is mediated by the constitutive androstane receptor.

Toxaphene was shown to increase liver tumor incidence in B6C3F1 mice following chronic dietary exposure. Preliminary evidence supported a role for the constitutive androstane receptor (CAR) in the mode of action of toxaphene-induced mouse liver tumors. However, these results could not rule out a role for the pregnane X receptor (PXR) in liver tumor formation. To define further the nuclear receptors involved in this study, we utilized CAR, PXR and PXR/CAR knockout mice (CAR-/- , PXR-/- and PXR-/- /CAR-/- ) along with the wild-type C57BL/6. In this study CAR-responsive genes Cyp3a11 and Cyp2b10 were induced in the liver of C57BL/6 (wild-type) mice by toxaphene (30-570-fold) (at the carcinogenic dose 320 ppm) and phenobarbital (positive control) (16-420-fold) following 14 days' dietary treatment. In contrast, in CAR-/- mice, no induction of these genes was seen following treatment with either chemical. Cyp3a11 and Cyp2b10 were also induced in PXR-/- mice with toxaphene and phenobarbital but were not changed in treated PXR-/- /CAR-/- mice. Similarly, induction of liver pentoxyresorufin-O-deethylase (CAR activation) activity by toxaphene and phenobarbital was absent in CAR-/- and PXR-/- /CAR-/- mice treated with phenobarbital or toxaphene. Ethoxyresorufin-O-deethylase (EROD, represents aryl hydrocarbon receptor activation) activity in CAR-/- mice treated with toxaphene or phenobarbital was increased compared with untreated control, but lower overall in activity in comparison to the wild-type mouse. Liver EROD activity was also induced by both phenobarbital and toxaphene in the PXR-/- mice but not in the PXR-/- /CAR-/- mice. Toxaphene treatment increased 7-benzyloxyquinoline activity (a marker for PXR activation) in a similar pattern to that seen with pentoxyresorufin-O-deethylase. These observations indicate that EROD and PXR activation are evidence, as expected, of secondary overlap to primary CAR receptor activation. Together, these results definitively show that activation of the CAR nuclear receptor is the mode of action of toxaphene-induced mouse liver tumors. Copyright © 2017 John Wiley & Sons, Ltd.

Fumio Matsumura--accomplishments at the University of California, Davis, and in the Sierra Nevada Mountains.

Fumio Matsumura joined the University of California, Davis, faculty in 1987 where he served as founding director of the Center for Environmental Health Sciences, associate director of the U.C. Toxic Substances Research and Teaching Program, and chair of the Department of Environmental Toxicology. He was an active affiliate with the NIEHS-funded Superfund Basic Research Program and the NIH Comprehensive Cancer Center. He was in many instances a primary driver or otherwise involved in most activities related to environmental toxicology at Davis, including the education of students in environmental biochemistry and ecotoxicology. A significant part of his broad research program was focused on the long range transport of chemicals such as toxaphene, PCBs and related contaminants used or released in California to the Sierra Nevada mountains, downwind of the urban and agricultural regions of the state. He hypothesized that these chemical residues adversely affected fish and wildlife, and particularly the declining populations of amphibians in Sierra Nevada streams and lakes. Fumio and his students and colleagues found residues of toxaphene and PCBs at higher elevations, an apparent result of atmospheric drift and deposition in the mountains. Fumio and his wife Teruko had personal interests in, and a love of the mountains, as avid skiers, hikers, and outdoor enthusiasts.

[Levels for toxaphene in three categories of animal originated food in Pearl River Delta area].

OBJECTIVE: To explore the present contamination status and profile of toxaphene in fish, chicken and meat samples in Pearl River Delta area. METHODS: Totally 60 independent samples including fish, chicken, meat were collected from Huizhou,Guangzhou,Dongguan,Jiangmen of Pearl River Delta area in Guangdong by purposive sampling from Oct 2010 to Jun 2011. Toxaphene monomer Parlar26, Parlar50 and Parlar62 levels in three various categories of foods were quantitatively and qualitatively analyzed referencing international authoritative standard methods by utilizing isotope dilution high-resolution gas chromatography/high-resolution double-focusing magnetic mass spectrometry (HRGC-HRMS). The levels of pollution and characteristics were compared. RESULTS: The concentration of toxaphene median(M) in fish, chicken and meat were 12.87, 5.8 and 1.89 ng/kg (gross weight) , respectively and the difference was significant(H = 14.29, P = 0.001). The toxaphene level in seafish (M = 32.07 ng/kg) was significantly higher than that in freshwater fish (M = 10.63 ng/kg, Z = -2.52, P = 0.012). The profile analysis showed that Parlar50 and Parlar26 were the predominant characteristic monomers in fish, which contributed about 44% (9.91/22.60, ng/kg) and 39% (8.89/22.60, ng/kg) to total concentration, and Parlar62 and Parlar26 were the two predominant congeners in poultry products, the two congeners accounted for 42% (5.03/11.90, ng/kg) and 38% (4.58/11.90, ng/kg). Furthermore Parlar26 and Parlar62 made up about 57% (3.45/6.08, ng/kg) and 26% (1.58/6.08, ng/kg) of total concentration in livestock meat respectively. CONCLUSION: Toxaphene levels in all tested food samples are far lower than the limited standard stipulated by the European Food Safety Authority (EFSA) in 2002, which was 0.1 mg/kg. The congener characteristic difference in the three categories food indicated that the different enrichment ability in Toxaphene in different animal species.

Scavenging amphipods: sentinels for penetration of mercury and persistent organic chemicals into food webs of the deep Arctic Ocean.

Archived specimens of the scavenging amphipod Eurythenes gryllus, collected from 2075 to 4250 m below the surface on five expeditions to the western and central Arctic Ocean between 1983 and 1998, were analyzed for total mercury (∑Hg), methyl mercury (MeHg), polychlorinated biphenyls (PCBs) and other industrial or byproduct organochlorines (chlorobenzenes, pentachloroanisole, octachlorostyrene), organochlorine pesticides (OCPs), and polybrominated diphenyl ethers (PBDEs). Median ∑Hg concentrations ranged from 70 to 366 ng g(-1) wet weight (ww). MeHg concentrations (3.55 to 23.5 ng g(-1) ww) accounted for 1.7 to 20.1% (median 3.7%) of ∑Hg. ∑Hg and MeHg were positively and significantly correlated with ww (∑Hg r(2) = 0.18, p = 0.0004, n = 63; MeHg r(2) = 0.42, p = 0.0004, n = 25), but not significantly with δ(13)C nor δ(15)N. Median concentrations of total persistent organic pollutants (POPs) ranged from 9750 to 156,000 ng g(-1) lipid weight, with order of abundance: ∑TOX (chlorobornanes quantified as technical toxaphene) > ∑PCBs > ∑DDTs > ∑chlordanes > ∑mirex compounds > ∑BDEs ∼ ∑chlorobenzenes ∼ octachlorostyrene > α-hexachlorocyclohexane ∼ hexachlorobenzene ∼ pentachloroanisole. Enantioselective accumulation was found for the chiral OCPs o,p'-DDT, cis- and trans-chlordane, nonachlor MC6 and oxychlordane. Lipid-normalized POPs concentrations were elevated in amphipods with lipid percentages ≤10%, suggesting that utilization of lipids resulted in concentration of POPs in the remaining lipid pool. Multidimensional Scaling (MDS) analysis using log-transformed physiological variables and lipid-normalized organochlorine concentrations distinguished amphipods from the central vs western arctic stations. This distinction was also seen for PCB homologues, whereas profiles of other compound classes were more related to specific stations rather than central-west differences.

Profiles and determinants of dicofol, endosulfans, mirex, and toxaphenes in breast milk samples from 10 prefectures in Japan.

Human exposure to persistent organic pollutants (POPs) is reflected by POP concentrations in breast milk. Many studies of POPs in breast milk have been performed in Japan, but insufficient information is available about some legacy POPs (e.g., mirex and toxaphenes, included in the Stockholm Convention in 2001) and novel POPs (e.g., dicofol and endosulfans, included in the Stockholm Convention in 2019 and 2011, respectively). In this study, dicofol, endosulfan, mirex, and toxaphene concentrations in breast milk from 10 prefectures in Japan were determined. The samples were collected between 2005 and 2010, before Stockholm Convention restrictions on endosulfans and mirex were implemented. Common POPs (e.g., polychlorinated biphenyls) were also analyzed to allow the contamination statuses to be compared. The α-endosulfan and ß-endosulfan concentrations were 0.26-13 and 0.012-0.82 ng/g lipid, respectively. The toxaphene #26 and #50 concentrations were <0.08-5.6 and < 0.1-8.5 ng/g lipid, respectively. The dicofol concentrations were <0.01-4.8 ng/g lipid. The mirex concentrations were <0.2-3.5 ng/g lipid. The α- and ß-endosulfan concentrations on a lipid weight basis negatively correlated with the lipid contents of the milk samples (ρ = -0.65, p < 0.01 for α-endosulfan; ρ = -0.58, p < 0.01 for ß-endosulfan). The toxaphene concentrations positively correlated with the lipid contents. The mirex concentrations positively correlated with the maternal age but negatively correlated with the maternal body mass index. No correlations between the dicofol concentrations and the factors were found. Principal component analysis divided the data into four groups, (1) chlordanes, dichlorodiphenyltrichloroethanes and related compounds, hexachlorobenzene, hexachlorocyclohexanes, hexachloroethane, and polychlorinated biphenyls, (2) endosulfans, (3) dicofol, dieldrin, and toxaphenes, and (4) bromodiphenyl ether 47. This indicated that bromodiphenyl ether 47, dicofol, endosulfans, and toxaphenes have different exposure routes or different kinetics to the other legacy POPs.

Characteristic molecular signature for early detection and prediction of persistent organic pollutants in rat liver.

Persistent organic pollutants (POPs) are degradation-resistant anthropogenic chemicals that accumulate in the food chain and in adipose tissue, and are among the most hazardous compounds ever synthesized. However, their toxic mechanisms are still undefined. To investigate whether characteristic molecular signatures can discriminate individual POP and provide prediction markers for the early detection of POPs exposure in an animal model, we performed transcriptomic analysis of rat liver tissues after exposure to POPs. The six different POPs (toxaphene, hexachlorobenzene, chlordane, mirex, dieldrin, and heptachlor) were administered to 11-week-old male Sprague-Dawley rats, and after 48 h of exposure, RNAs were extracted from liver tissues and subjected to rat whole genome expression microarrays. Early during exposure, conventional toxicological analysis including changes in the body and organ weight, histopathological examination, and blood biochemical analysis did not reflect any toxicant stresses. However, unsupervised gene expression analysis of rat liver tissues revealed in a characteristic molecular signature for each toxicant, and supervised analysis identified 2708 outlier genes that discerned the POPs exposure group from the vehicle-treated control. Combination analysis of two different multiclassifications suggested 384 genes as early detection markers for predicting each POP exposure with 100% accuracy. The data from large-scale gene expression analysis of a different POP exposure in rat model suggest that characteristic expression profiles exist in liver hepatic cells and multiclassification of POP-specific molecular signatures can discriminate each toxicant at an early exposure time. The use of these molecular markers may be more widely implemented in combination with more traditional techniques for assessment and prediction of toxicity exposure to POPs from an environmental aspect.

Polybrominated diphenyl ethers, toxaphenes, and other halogenated organic pollutants in great blue heron eggs.

The great blue heron (Ardea herodias) has been used as a bioindicator of the state of the St. Lawrence River (Québec, Canada) since 1996. At 5-year intervals, selected breeding colonies along the River and its estuary are visited to estimate reproductive success and determine levels of contamination. Brominated flame retardants are found in many ecosystems and are increasing in concentration in the Great Lakes, which is the source of much of the water for the St. Lawrence River. In 2001 and 2002, in addition to polychlorinated biphenyls (PCBs) and chlorinated pesticides, the levels of polybrominated diphenyl ethers (PBDEs), polychlorinated bornanes (toxaphene) congeners and non-ortho-substituted PCBs were measured for the first time in pools of great blue heron eggs. The PBDE levels in great blue heron eggs (70-1,377 ng/g wet wt) were comparable to those measured in herring gull (Larus argentatus) eggs from the Great Lakes. Toxaphene was detected in great blue heron eggs at levels comparable to those of other major chlorinated pesticides. Major toxaphene congeners were octachlorobornane P44 and the nonachlorobornane P50. Environ.

The organochlorine pesticide toxaphene reduces non-mitochondrial respiration and induces heat shock protein 70 expression in early-staged zebrafish (Danio rerio).

Toxaphene is a restricted-use pesticide produced by reacting chlorine gas with camphene. It was heavily used as a pesticide for agricultural purposes in the 1960-1970s, but despite being banned >30 years ago, it can remain elevated in the soil due to its resistance to metabolic degradation; this has led to longstanding concerns about elevated levels of toxaphene and other organochlorine pesticides (OCPs) in the environment. The objective of this study were to determine the effects of waterborne exposure to toxaphene on early life stages of zebrafish. Based on the LC50, zebrafish embryos were exposed to control (embryo rearing media or DMSO) or to one dose of toxaphene ranging between 0.011 and 111.1 µg/mL from 6 h post fertilization (hpf) up to 120 hpf. Significant mortality and hatch time delays were observed in embryos exposed to toxaphene (at or above 0.11 and 1.11 µg/mL, depending on the assay). Higher prevalence of deformities was noted at higher doses (≥0.011 µg/mL), and these included pericardial edema and skeletal deformities. As energy production is important for normal development, mitochondrial bioenergetics were assessed in embryos following toxaphene exposure. Embryos exposed to 11.1 or 111 µg/mL toxaphene for 24 h showed lower non-mitochondrial respiration (~30%) compared to both solvent and no treatment controls. Expression of transcripts related to oxidative damage responses and apoptosis were measured and heat shock protein 70 was significantly increased with 111 µg/mL toxaphene (14.5 fold), while the expression levels of caspase 3, caspase 9, and superoxide dismutase 1 were not changed. These data demonstrate that developmental deformities induced by toxaphene include pericardial edema and skeletal deformity, and that toxaphene can affect oxidative phosphorylation in early staged zebrafish.

Transcriptome and physiological effects of toxaphene on the liver-gonad reproductive axis in male and female largemouth bass (Micropterus salmoides).

Toxaphene is an organochlorine pesticide and environmental contaminant that is concerning due to its atmospheric transport and persistence in soil. In Florida, toxaphene and other organochlorine pesticides were used heavily in agriculture on the north shore of Lake Apopka and they are still detectable in soil. Wild largemouth bass that inhabit the lake and the marshes along the north shore have been exposed to a variety of organochlorine pesticides including dieldrin, methoxychlor, and p,p'-DDE, among others. While these other organochlorine pesticides have been studied for their endocrine disrupting effects in largemouth bass, there is little information for toxaphene. In this study, male and female largemouth bass were given food containing 50 mg/kg toxaphene for almost 3 months, to achieve tissue levels similar to those found in fish at Lake Apopka. Sex-specific toxicity was then evaluated by measuring various reproductive endpoints and transcriptomic changes. In females, gonadosomatic index showed a trend towards reduction (p = 0.051) and plasma vitellogenin was reduced by ~40% relative to controls. However plasma levels of 17ß-estradiol and testosterone were not perturbed by toxaphene exposure. These data suggest that toxaphene does not act as a weak estrogen as many other organochlorine pesticides do, but rather appears to be acting as an antiestrogen in female fish. There were no obvious changes in the gonadosomatic index and plasma hormones in male bass. However, ex vivo explant experiments revealed that toxaphene prevented human chorionic gonadotropin-stimulated testosterone production in the testis. This suggested that toxaphene had anti-androgenic effects in males. Subsequent transcriptomic analyses of the testis revealed that androgen receptor/beta-2-microglobulin signaling was up-regulated while insulin-related pathways were suppressed with toxaphene, which could be interpreted as a compensatory response to androgen suppression. In the male liver, the transcriptome analysis revealed an overwhelming suppression in immune-related signaling cascades (e.g. lectin-like receptor and ITSM-Containing Receptor signaling, CD16/CD14 Proinflammatory Monocyte Activation, and CD38/CD3-JUN/FOS/NF-kB Signaling in T-cell Proliferation). Overall, this study showed that toxaphene induced sex-specific effects. The transcriptomic and physiological responses observed can contribute to the development of adverse outcome pathways for toxaphene exposure in fish.

Polychlorobornane components of toxaphene: structure-toxicity relations and metabolic reductive dechlorination.

2,2,5-endo,6-exo,8,9,10-Heptachlorobornane and four derivatives of this heptachlorobornane, with an additional chlorine atom at position 3-exo,8,9, or 10, account for a major portion of the acute toxicity of toxaphene and for up to 23 percent of toxaphene composition as analyzed by open tubular column gas-liquid chromatography with an electron capture detector. Both in several organisms and model environmental systems and on photolysis, this heptachlorobornane undergoes facile reductive dechlorination at the geminal-dichloro group and sometimes dehydrochlorination.

Toxaphene insecticide: a complex biodegradable mixture.

Adsorption and gas-liquid chromatography separate toxaphene into at least 175 polychlorinated 10-carbon compounds including Cl(6), Cl(7), Cl(8), Cl(9), and Cl(10) derivatives. One toxic component is 2,2,5-endo,6-exo,8,9,10-heptachlorobornane. Rats metabolically dechlorinate toxaphene, removing about half of the chlorine from the technical insecticide and from each of seven subfractions of varying composition and toxicity.

Toxaphene, a complex mixture of polychloroterpenes and a major insecticide, is mutagenic.

Toxaphene, the most widely used chlorinated insecticide, is mutagenic in the Salmonella test without requiring liver homogenate for activity. This insecticide is a complex mixture (more than 177 polychloroterpenes) with carcinogenic activity in rodents. Some but not all of the mutagenic components are easily separated from the insecticidal ingredients.

Synergistic activation of estrogen receptor with combinations of environmental chemicals.

Certain chemicals in the environment are estrogenic. The low potencies of these compounds, when studied singly, suggest that they may have little effect on biological systems. The estrogenic potencies of combinations of such chemicals were screened in a simple yeast estrogen system (YES) containing human estrogen receptor (hER). Combinations of two weak environmental estrogens, such as dieldrin, endosulfan, or toxaphene, were 1000 times as potent in hER-mediated transactivation as any chemical alone. Hydroxylated polychlorinated biphenyls shown previously to synergistically alter sexual development in turtles also synergized in the YES. The synergistic interaction of chemical mixtures with the estrogen receptor may have profound environmental implications. These results may represent a previously uncharacterized level of regulation of estrogen-associated responses.

Development of a novel high volume band compression injector for the analysis of complex samples like toxaphene pesticide.

A new type of injector has been developed for gas chromatographic analysis. The injector has high volume and band compression (HVBC) capabilities useful for the analysis of complex samples. The injector consists essentially of a packed liner operated at room temperature while a narrow heated zone is used to axially scan the liner selectively desorbing the compounds of interest. The scanning speed, distance and temperature of the zone are precisely controlled. The liner is connected to an interface which can vent the solvent or any undesirable compounds, and transfer the analytes to an analytical column for separation and quantification. The injector is designed to be compatible with injection volumes from 1 to more than 250microL. At a low sample volume of 1microL, the injector has competitive performances compared to those of the "on-column" and "split/splitless" injectors for the fatty acid methyl esters and toxaphene compounds tested. For higher volumes, the system produces a linear response according to the injected volume. In this explorative study, the maximum volume injected seems to be limited by the saturation of the chromatographic system instead of being defined by the design of the injector. The HVBC injector can also be used to conduct "in situ" pretreatment of the sample before its transfer to the analytical column. For instance, a toxaphene sample was successively fractionated, using the HVBC injector, in six sub-fractions characterized by simpler chromatograms than the chromatogram of the original mixture. Finally, the ability of the HVBC injector to "freeze" the separation in time allowing the analyst to complete the analysis at a later time is also discussed.

Toxaphene analysis in Great Lakes fish: a comparison of GC-EI/MS/MS and GC-ECNI-MS, individual congener standard and technical mixture for quantification of toxaphene.

Toxaphene is considered to be a problematic organochlorine pollutant because of its bioaccumulation potential and persistence in aquatic environments. In this study, whole lake trout and walleye composites were used to evaluate two analytical techniques for total toxaphene and selected congener analysis. The efficacy of using gas chromatography electron ionization tandem mass spectrometry (GC-EI/MS/MS) and electron capture negative ionization mass spectrometry (GC-ECNI-MS) were compared. Although the sensitivity using GC-ECNI-MS was approximately five times greater than GC-EI/MS/MS, the latter provided more consistent inter-Parlar relative response factors (RRF). When using technical calibration mixtures, these results suggest a more accurate total toxaphene measurement was obtained using the GC-EI/MS/MS method. Total toxaphene concentrations in lake trout composites from both methods were highly correlated (R(2) = 0.985) with the MS/MS concentrations approximately half of those determined by ECNI, suggesting systematic high bias in toxaphene concentrations when measured using GC-ECNI.