Effects of three biodiesels and a low sulfur diesel in male rats--a pilot 4-week oral study.

Because of the accessible and renewable nature of feedstock and the potential for the reduction of harmful combustion emissions and greenhouse gases, biodiesels have received increasing interest as an alternate fuel. Oral exposure to biodiesels is a concern because of contact during refuelling, accidental ingestion and exposure through ground water contamination. Although biodiesels from various feedstock are in use commercially and experimentally, very little is known about their potential adverse effects and no data is available on their potential for ground water contamination. A study was performed on male rats following oral treatment with experimental biodiesels (dissolved in corn oil) derived from canola oil (Bio-C), soy oil (Bio-S) and fish oil (Bio-F), at 500 mg/kg body weight/day, 5 days per week, for 4 weeks. Separate groups of animals were treated with low sulfur diesel (LSD) for comparison purpose, and with corn oil alone to serve as control. The potential for ground water contamination by biodiesels was investigated by the preparation of water-accommodated fractions (WAF) followed by gas chromatographic analysis. WAF from Bio-F and Bio-S was found to have the highest level of dichloromethane extractable materials. Gas chromatographic analysis indicated that the extractable materials from biodiesels contained much higher proportion of C15-C30 materials than LSD. Increased liver weight was observed in animal treated with Bio-C, Bio-S and LSD and decreased thymus weight was found in those treated with Bio-S. Histopathological changes typical of male-rat specific hyaline-droplet nephropathy were detected in kidney tubules of animals treated with LSD, Bio-S and Bio-C. Mild adaptive changes were observed in thyroids of animals treated with LSD, Bio-S and Bio-F. Clinical chemical and biochemical changes were confined to Bio-S and LSD treated rats and included elevation in some hepatic phase-I and phase-II drug metabolizing enzymes and hepatic palmitoyl Co-A oxidase, and elevated urinary concentrations of ascorbic acid and albumin. At the given dose level of 500 mg/kg bw/day, the overall treatment-related effects of biodiesels and LSD are mild, and the severity of the treatment effects may be ranked as: LSD>Bio-S>Bio-C>Bio-F. Considered together with the presence of a higher level of water extractable materials, Bio-S may be more of a concern for potential human health than Bio-C and Bio-F in an oral exposure scenario. Further studies are needed to identify and characterize the constituents contributing to the treatment-related effects specific to these experimental biodiesels.

Headspace solid-phase microextraction for the determination of volatile and semi-volatile pollutants in water and air.

In this work we report the use of solid-phase microextraction (SPME) to extract and concentrate water-soluble volatile as well as semi-volatile pollutants. Both methods of exposing the SPME fibre were utilised: immersion in the aqueous solution (SPME) and in the headspace over the solution (HSSPME). The proposed HSSPME procedure was compared to conventional static headspace (HS) analysis for artificially spiked water as well as real water samples, which had been, equilibrated with various oil and petroleum products. Both techniques gave similar results but HSSPME was much more sensitive and exhibited better precision. Detection limits were found to be in the sub-ng/ml level, with precision better than 5% R.S.D. in most cases. To evaluate the suitability of SPME for relatively high contamination level analysis, the proposed HSSPME method was applied to the screening of run-off water samples that had heavy oil suspended in them from a tire fire incident. HSSPME results were compared with liquid--liquid extraction. Library searches were conducted on the resulting GC-MS total ion chromatograms to determine the types of compounds found in such samples. Both techniques found similar composition in the water samples with the exception of alkylnaphthalenes that were detected only by HSSPME. A brief study was carried out to assess using SPME for air monitoring. By sampling and concentrating the volatile organic compounds in the coating of the SPME fibre without any other equipment, this new technique is useful as an alternative to active air monitoring by means of sampling pumps and sorbent tubes.

Characterization and identification of a "mystery" oil spill from Quebec (1999).

This paper describes a case study in which advanced chemical fingerprinting and data interpretation techniques were used to characterize the chemical compositions and determine the source of an unknown spilled oil from Quebec. On 28 February 1999, significant amounts of oil was reported on the river banks of St. Laurence River in front of a company named "Thermex" (in a town - Beauharnois, Quebec, about 50 km northwest of Montreal). The spilled oil was suspected to be released from a nearby factory. In response to this specific site investigation needs, a tiered analytical approach using GC-MS and GC-flame ionization detection was applied. A variety of diagnostic ratios of "source-specific marker" compounds, in particular isomers of biomarkers and alkylated series of polycyclic aromatic hydrocarbons within the same alkylation groups, were determined and analyzed. The hydrocarbon analysis results reveal the following: (1) the spilled oil is very "specific", and is significantly different from most crude oils in chemical composition; (2) the oil in samples come from the same source, however, the spill sample 2569 was identified to contain a small amount (approximately 10%) of diesel; (3) the spilled oil was relatively "fresh", its chemical composition has not undergone significant alteration yet; (4) the spilled oil showed unusually high concentration of the US Environmental Protection Agency priority polycyclic aromatic hydrocarbons (PAHs). The "Pyrogenic Index" values were determined to be as high as 0.11-0.13, significantly higher than crude oils (<0.010) and heavy Bunker type fuels (0.015-0.060). This indicates significant contribution of PAH composition from pyrogenic components; (5) biomarkers were also detected, but their concentrations were unusually low in comparison to most crude oils.

Characterization and source identification of hydrocarbons in water samples using multiple analytical techniques.

This paper describes a case study in which multiple analytical techniques were used to identify and characterize trace petroleum-related hydrocarbons and other volatile organic compounds in groundwater samples collected in a bedrock aquifer exploited for drinking water purposes. The objective of the study was to confirm the presence of gasoline and other petroleum products or other volatile organic pollutants in those samples in order to assess the respective implication of each of the potentially responsible parties to the contamination of the aquifer. In addition, the degree of contamination at different depths in the aquifer was also of interest. The analytical techniques used for analyses of water samples included gas chromatography-mass spectrometry (GC-MS) and capillary GC with flame-ionization detection, solid-phase microextraction and headspace GC-MS techniques. Chemical characterization results revealed the following: (1) The hydrocarbons in sample A (near-surface groundwater, 0-5 m) were clearly of two types, one being gasoline and the other a heavy petroleum product. The significant distribution of five target petroleum-characteristic alkylkated polycyclic aromatic hydrocarbon homologues and biomarkers confirmed the presence of another heavy petroleum product. The concentrations of the TPHs (total petroleum hydrocarbons) and BTEX (collective name of benzene, toluene, ethylbenzene, and p-, m-, and o-xylenes) were determined to be 1070 and 155 microg/kg of water for sample A, respectively. (2) The deepest groundwater (sample B, collected at a depth ranging between 15 and 60 m) was also contaminated, but to a much lesser degree. The concentrations of the TPH and BTEX were determined to be only 130 and 2.6 microg/kg of water for sample B, respectively. (3) The presence of a variety of volatile chlorinated compounds to the groundwater was also clearly identified.

Characterization and identification of the Detroit River mystery oil spill (2002).

In this paper, a case study of the Detroit River mystery oil spill (2002) is presented that demonstrates the utility of detailed and integrated oil fingerprinting in investigating unknown or suspected oil spills. The detailed diagnostic oil fingerprinting techniques include determination of hydrocarbon groups and semi-quantitative product screening, analysis of oil-characteristic biomarkers and the extended suite of parent and alkylated polycyclic aromatic hydrocarbons (PAHs), and quantitative determination of a variety of diagnostic ratios of "source-specific marker" compounds. The detailed chemical fingerprinting data and results highlight the followings: (1) The spill samples were largely composed of used lube oil mixed with smaller portion of diesel fuel. (2) The diesel in the samples had been weathered and degraded. (3) Sample 3 collected from N. Boblo Island on 14 April was more weathered (most probably caused by more evaporation and water-washing) than samples 1 and 2. (4) All fingerprinting results clearly demonstrated oils in three samples were the same, and they came from the same source. (5) Most PAH compounds were from the diesel portion in the spill samples, while the biomarker compounds were largely from the lube oil. (6) Input of pyrogenic PAHs to the spill samples was clearly demonstrated. The pyrogenic PAHs were most probably produced from combustion and motor lubrication processes, and the lube oil in these spill samples was waste lube oil.

Long-term fate and persistence of the spilled metula oil in a marine salt marsh environment degradation of petroleum biomarkers.

Three coastal sites, heavily oiled from the 1974 Metula oil spill in the Strait of Magellan [two are salt marshes (East and West) and the third, an intertidal asphalt pavement], were examined during May 1998. Complete 'total oil analyses' were performed on the oil samples collected from these sites. Chemical fingerprinting data reveal, except for those samples from the East Marsh untreated plots which were only lightly to moderately weathered, that the spilled oil has undergone significant alteration in chemical composition after 24 years. There are no fundamental differences between the heavily weathered West Marsh and treated East Marsh samples. However, the effect of experimental filling action conducted in 1993 has been to substantially promote plant recolonization. The asphalt pavement samples indicate extremely high degradation of oil hydrocarbons, evidenced by a complete loss of n-alkanes from n-C8 to n-C41 and by depletion of greater than 98% of the alkylated polycyclic aromatic hydrocarbon homologues. Even the most refractory biomarker compounds showed some degree of biodegradation. The biomarkers were generally degraded in the declining order of importance as follows: diasteranes>C27 steranes>tricyclic terpanes>pentacyclic terpanes>norhapanes approximately C29-alphabetabeta-steranes.

Comparison of oil composition changes due to biodegradation and physical weathering in different oils.

The well-characterized Alberta Sweet Mixed Blend oil and several other oils which are commonly transported in Canada were physically weathered and then incubated with a defined microbial inoculum. The purpose was to produce quantitative data on oil components and component groups which are more susceptible or resistant to biodegradation, and to determine how oils rank in relation to each other in terms of biodegradation potential. The biodegraded oils were characterized by quantitative determination of changes in important hydrocarbon groups including the total petroleum hydrocarbons, total saturates and aromatics, and also by quantitation of more than 100 individual target aliphatic, aromatic and biomarker components. The study reveals a pattern of distinct oil composition changes due to biodegradation, which is significantly different from the pattern due to physical or short-term weathering. It is important to be able to distinguish between these two forms of loss, so that loss due to weathering is not interpreted as loss due to biodegradation in the laboratory or in the field. Based on these findings, the oil composition changes due to biodegradation can be readily differentiated from those due to physical weathering. To rank the tested oils with respect to biodegradability, losses in total petroleum hydrocarbons and aromatics were used to calculate biodegradation potential indices, employing equations proposed by Environment Canada and the US National Oceanic and Atmospheric Administration. The different methods produced very similar biodegradation trends, confirming that patterns of oil biodegradability do exist.

Headspace solid phase microextraction (HSSPME) for the determination of volatile and semivolatile pollutants in soils.

We have investigated the use of headspace solid phase microextraction (HSSPME) as a sample concentration and preparation technique for the analysis of volatile and semivolatile pollutants in soil samples. Soil samples were suspended in solvent and the SPME fibre suspended in the headspace above the slurry. Finally, the fibre was desorbed in the Gas Chromatograph (GC) injection port and the analysis of the samples was carried out. Since the transfer of contaminants from the soil to the SPME fibre involves four separate phases (soil-solvent-headspace and fibre coating), parameters affecting the distribution of the analytes were investigated. Using a well-aged artificially spiked garden soil, different solvents (both organic and aqueous) were used to enhance the release of the contaminants from the solid matrix to the headspace. It was found that simple addition of water is adequate for the purpose of analysing the target volatile organic chemicals (VOCs) in soil. The addition of 1 ml of water to 1 g of soil yielded maximum response. Without water addition, the target VOCs were almost not released from the matrix and a poor response was observed. The effect of headspace volume on response as well as the addition of salt were also investigated. Comparison studies between conventional static headspace (HS) at high temperature (95 degrees C) and the new technology HSSPME at room temperature ( approximately 20 degrees C) were performed. The results obtained with both techniques were in good agreement. HSSPME precision and linearity were found to be better than automated headspace method and HSSPME also produced a significant enhancement in response. The detection and quantification limits for the target VOCs in soils were in the sub-ng g(-1) level. Finally, we tried to extend the applicability of the method to the analysis of semivolatiles. For these studies, two natural soils contaminated with diesel fuel and wood preservative, as well as a standard urban dust contaminated with polyaromatic hydrocarbons (PAHs) were tested. Discrimination in the response for the heaviest compounds studied was clearly observed, due to the poor partition in the headspace and to the slow kinetics of all the processes involved in HSSPME.

An evaluation of field total petroleum hydrocarbon (TPH) systems.

An evaluation of several field kits and petroleum hydrocarbon measuring systems was conducted. The field kits were the immunoassay based EnviroGard petroleum fuels in soil test kit (EnviroGard, Millipore Canada, Mississauga, Ont., Canada), the turbidimetric based PetroFlag hydrocarbon test kit for soil (Dexsil, Hamden, CT, USA), a DR/2000 field kit (Hach Company, Loveland CO, USA) employing colorimetric test procedures and a total organic carbon (TOC) analysis instrument (Dohrmann Division, Rosemount Analytical Inc., Santa Clara, CA, USA) using oxidation principles. These procedures were compared to the traditional technique of extraction of the petroleum hydrocarbons using trichlorotrifluoroethane (Freon 113) as the solvent and subsequent infrared (IR) analysis using a portable fixed wavelength analyzer (Buck Scientific, East Norwalk, CT, USA). The EnviroGard kit was affected by the sample matrix. The soil type and the presence or lack thereof specific chemical components affected the capability to detect the petroleum hydrocarbon concentration. The PetroFlag soil test kit tended to generate results higher than the accepted concentration. The IR method was better capable of producing results similar to the expected concentration values of the prepared samples. Results indicate that the total organic carbon analysis technique evaluated is best suited for samples containing dissolved hydrocarbons in water and is not a preferred procedure for water samples containing dispersed or floating oil. At low concentrations of 10ppm and less, the TOC method and IR method have concentration values within a few parts-per-million (ppm) of each other, however, an examination of the trends in the results for all samples shows no similarity. This would indicate that the traditional extraction and infrared method and the total organic carbon method are not measuring the same parameter.Finally, the colorimetric field kit was capable of quantifying the concentration of oil in water samples within limits. The results from the oil-in-water method built into the unit at the factory were not comparable with analysis carried out by the infrared technique. With specific methods for each oil incorporated into the spectrophotometer, the comparability of data increased significantly. Results generated by the kit are dependent upon the color and amount of the oil in the sample. The kit is best suited for dark colored oils and the water samples with concentrations in the range of 10 to 85ppm by weight.

On site PCB analysis in support of a transformer rebuilding project.

In December 1997, Emergencies Science Division (ESD) was contracted by Natural Resources Canada (NRCAN) to perform on-site analyses in support of a transformer-rebuilding project at Sault Ste-Marie, Ont. Using a gas chromatograph with electron capture detector (GC/ECD) mounted in a mobile laboratory, PCB analyses were conducted on the original transformer oil, surface wipes, Varsol rinsing of the transformer tank interior and cooling fins. To assess the efficiency and validity of the decontamination process, PCB contamination was monitored closely on the rinse solvent. Surface wipe samples after wash down showed surface concentration of several hundred microg Aroclor 1254/100 cm(2), well below the acceptable limit of 8000 microg/100 cm(2). Because of the relatively large percentage of the internal surface area, the fin banks had to be rinsed exhaustively to meet the decontamination criteria. Final rinses of each of the seven fin banks of transformer 1 still showed presence of PCB, ranging from 80 to 590 ppm (microg/ml) with a mean value of 280 ppm. Upon completion of rebuilding, analysis of the R-Temp retro fill fluid showed 5 ppm at the initial power-up, increasing slightly to 16 ppm after 1 year of operation, which was far below the regulatory limit 50 ppm. The second transformer, by comparison, had a lower mean concentration of 54 ppm in the final fin rinse during decontamination. However, the backfill R-Temp showed an initial concentration of 38 ppm and remained essentially unchanged at 32 ppm after approximately 10 months of operation. Extensive comparison of GC and the quick test Clor-N-Oil kit were also carried out and showed generally good agreement. The use of an on-site GC was crucial in providing rapid and accurate analysis on-site, thus, enabling quick modifications to the decontamination strategies in order to meet the target PCB level. For projects of this nature, a GC/ECD was far superior to quick test kits by providing the selectivity and sensitivity for the diverse nature of the sample media.

Field fluorometers as dispersed oil-in-water monitors.

A laboratory study of the Turner Instrument flow-through models 10AU and 10 fluorometers was conducted to review their ability to measure real-time oil-in-water concentrations, to compare the results to other total petroleum hydrocarbon (TPH) procedures and to improve the understanding of the relationship of the fluorescence to the chemical composition of the oils. Comparison of the fluorometer results to standard infrared and gas chromatography laboratory procedures showed all methods capable of detecting and differentiating between small changes in oil concentration. The infrared and gas chromatography generated similar values while the fluorometer values were of the same order of magnitude but typically 20-80% higher. The chemical composition of the oils was determined by gas chromatographic techniques and compared to the signal outputs of the fluorometers. It was found that the fluorometer data could not be directly linked to the concentration of any specific aromatic hydrocarbon such as naphthalene or to the sum of the polycyclic aromatic hydrocarbon (PAH) compounds. Evidence suggests that the fluorescence signal is generated by a combination of PAH compounds. Also, the response of the fluorometers may also be influenced by the presence of volatile aromatic compounds such as benzene, toluene, ethyl benzene and xylene (BTEX) and C3-benzenes (BTEX + C3B) in combination with the PAH compounds.

Solvent vapour monitoring in work space by solid phase micro extraction.

Solid phase micro extraction (SPME) is a fast, solvent-less alternative to conventional charcoal tube sampling/carbon disulfide extraction for volatile organic compounds (VOC). In this work, SPME was compared to the active sampling technique in a typical lab atmosphere. Two different types of fibre coatings were evaluated for solvent vapour at ambient concentration. A general purpose 100 microm film polydimethylsiloxane (PDMS) fibre was found to be unsuitable for VOC work, despite the thick coating. The mixed-phase carboxen/PDMS fibre was found to be suitable. Sensitivity of the SPME was far greater than charcoal sorbent tube method. Calibration studies using typical solvent such as dichloromethane (DCM), benzene (B) and toluene (T) showed an optimal exposure time of 5 min, with a repeatability of less than 20% for a broad spectrum of organic vapour. Minimum detectable amount for DCM is in the range of 0.01 microg/l (0.003 ppmv). Variation among different fibres was generally within 30% at a vapour concentration of 1 microg DCM/l, which was more than adequate for field monitoring purpose. Adsorption characteristics and calibration procedures were studied. An actual application of SPME was carried out to measure background level of solvent vapour at a bench where DCM was used extensively. Agreement between the SPME and the charcoal sampling method was generally within a factor of two. No DCM concentration was found to be above the regulatory limit of 50 ppmv.

Review of behaviour of oil in freezing environments.

The current knowledge of the physical fate and behaviour of crude oil and petroleum products spilled in Arctic situations is reviewed. The fate and final deposition of oil in marine conditions is presented as based on the extant literature. Spreading models were evaluated for oil on ice, under ice, in snow, in brash ice, and between blocks of ice. Models of oil transport under sheet and broken ice were considered, both for sea and river conditions. The ability of ice sheets to trap oil is discussed in relation to oil storage capacity. The effects of oil on a growing ice sheet were examined, both in terms of ice formation and the thermal effects of oil inclusions in ice. The migration of oil through ice was reviewed, focussing primarily on the movement through brine channels. The effects of oil on the surface of ice were considered, with emphasis on the effects of surface pools on ice melt. Similar consideration was given to the effects of oil on snow on the surface of ice. The few quantitative studies of oil in open and dynamic ice conditions are reviewed. Observations of intentional small-scale spills in leads and ice fields are reviewed and compared with observations from real spills. The conditions under which "oil pumping" from leads occurs were quantified. The most common ultimate fate of oil in an ice field is to be released onto the water surface.

Biodegradation of propionitrile by Klebsiella oxytoca immobilized in alginate and cellulose triacetate gel.

A microbial process for the degradation of propionitrile by Klebsiella oxytoca was studied. The microorganism, K. oxytoca, was isolated from the discharged wastewater of metal plating factory in southern Taiwan and adapted for propionitrile biodegradation. The free and immobilized cells of K. oxytoca were then examined for their capabilities on degrading propionitrile under various conditions. Alginate (AL) and cellulose triacetate (CT) techniques were applied for the preparation of immobilized cells. The efficiency and produced metabolic intermediates and end-products of propionitrile degradation were monitored in bath and continuous bioreactor experiments. Results reveal that up to 100 and 150 mM of propionitrile could be removed completely by the free and immobilized cell systems, respectively. Furthermore, both immobilized cell systems show higher removal efficiencies in wider ranges of temperature (20-40 degrees C) and pH (6-8) compared with the free cell system. Results also indicate that immobilized cell system could support a higher cell density to enhance the removal efficiency of propionitrile. Immobilized cells were reused in five consecutive degradation experiments, and up to 99% of propionitrile degradation was observed in each batch test. This suggests that the activity of immobilized cells can be maintained and reused throughout different propionitrile degradation processes. A two-step pathway was observed for the biodegradation of propionitrile. Propionamide was first produced followed by propionic acid and ammonia. Results suggest that nitrile hydratase and amidase were involved in the degradation pathways of K. oxytoca. In the continuous bioreactor, both immobilized cells were capable of removing 150 mM of propionitriles completely within 16h, and the maximum propionitriles removal rates using AL and CT immobilized beads were 5.04 and 4.98 mM h(-1), respectively. Comparing the removal rates obtained from batch experiments with immobilized cells (AL and CT were 1.57 and 2.18 mM h(-1) at 150 mM of propionitrile, respectively), the continuous-flow bioreactor show higher potential for practical application.

Forensic fingerprinting of diamondoids for correlation and differentiation of spilled oil and petroleum products.

Diamondoids (adamantanes and diamantanes) are rigid, three-dimensionallyfused cyclohexyl-ring alkane compounds that can be found in almost all crude oils and in most petroleum products. Forforensic environmental investigations, the most commonly used biomarkers are high molecular weight (MW) tri- to pentacyclic terpanes and steranes. Most of these high MW biomarkers, however, are removed from the original crude oil feedstocks during the refining processes, while smaller biomarkers including diamondoids are concentrated in petroleum products. Fingerprinting diamondoids could thus provide another diagnostic means for correlation and differentiation of spilled oils and be particularly valuable for light to midrange distillates, such as jet and diesel fuels, the source of which may be difficult to identify using routine biomarker techniques. In this work, a reliable GC-MS analytical method has been developed for characterization and quantitation of diamondoids. The method detection limits for five target diamondoids were determined to be in the range of 0.06-0.14 microg/g oil. Distributions of diamondoids in over 100 different oils and refined products were quantitatively compared. The concentrations of four groups of target biomarkers were found, in general, to decrease in the order of sesquiterpanes > terpanes and steranes > adamantanes > diamantanes in both crude oils and refined products. A number of indices of admantanes and diamantanes have been developed and assessed as source indicators using their diagnostic powers (DP). The effects of evaporative weathering and biodegradation on alteration of diamondoid distributions have been quantitatively investigated. Finally, a spill case study by statistical evaluation of diagnostic ratios using the "two-tailed" Student's tapproach is presented to illustrate the unique utility of diamondoids for correlation and differentiation of unknown spilled diesels.

Solid-phase microextraction and headspace solid-phase microextraction for the determination of polychlorinated biphenyls in water samples.

A solid-phase microextraction (SPME) method has been developed for the quantification of polychlorinated biphenyls (PCBs) in water samples. Parameters such as sampling time, volume of water, volume of headspace, temperature, addition of salts, and agitation of the sample were studied. Because the time for reaching equilibrium between phases takes several hours or days, depending on the experimental conditions, it was necessary to work in nonequilibrium conditions to keep the sample analysis to a reasonable time. The possibility of sampling the headspace over the water sample (HSSPME), instead of immersing the fiber into the water (SPME), was also investigated, and despite the low partition of PCB into the headspace, HSSPME offered higher sensitivity than SPME at 100 °C. The adsorption kinetics for SPME at room temperature, SPME at 100 °C, and HSSPME at 100 °C were investigated and compared. The proposed HSSPME method exhibits excellent linearity and sensitivity. The detection limit was in the sub-ng/L level. This method has been applied to a real industrial harbor water and compared with liquid-liquid extraction. Both techniques offered similar results, but HSSPME was much more sensitive and considerably faster, by eliminating all the manual process intensive sample workup, and reduces solvent consumption entirely. The only drawback was that matrix effects were observed, but with the addition of deuterated surrogates to the sample or the use of a standard addition calibration, accurate quantification can be achieved.