Interlaboratory evaluation of endotoxin analyses in agricultural dusts--comparison of LAL assay and mass spectrometry

Endotoxin exposure is associated with wheeze and asthma morbidity, while early life exposure may reduce risk of allergy and asthma. Unfortunately, it is difficult to compare endotoxin results from different laboratories and environments. We undertook this study to determine if lipopolysaccharide (LPS) extraction efficiency could account for differences among laboratories. We generated and collected aerosols from chicken and swine barns, and corn processing. We randomly allocated side-by-side filter samples to five laboratories for Limulus assay of endotoxin. Lyophilized aliquots of filter extracts were analyzed for 3-hydroxy fatty acids (3-OHFAs) as a marker of LPS using gas chromatography-mass spectrometry. There were significant differences in endotoxin assay and GC-MS (LPS) results between laboratories for all dust types (p < 0.01). Patterns of differences between labs varied by dust type. Relationships between assay and GC/MS results also depended on dust type. The percentages of individual 3-OHFA chain lengths varied across labs (p < 0.0001) suggesting that each lab recovered a different fraction of the LPS available. The presence of large amounts of particle associated LPS and absence of a freezing thawing cycle were associated with lower correlations between LPS and bioactivity, consistent with an absence of Limulus response to cell-bound endotoxin. These data suggest that extraction methods affect endotoxin measurements. The LAL methods may be most suitable when comparing exposures within similar environments; GC-MS offers additional information helpful in optimizing sample treatment and extraction. GC-MS may be of use when comparing across heterogeneous environments and should be considered for inclusion in future studies of human health outcomes.     

The Variability of Estimates of Variance, and Its Effect on Power Analysis in Monitoring Design

Power analysis can be a valuable aid in the design of monitoringprograms. It requires an estimate of variance, which may come from a pilot study or an existing study in a similar habitat. For marine benthic infauna, natural variation in abundances canbe considerable, raising the question of reliability of varianceestimates. We used two existing monitoring programs to generatemultiple estimates of variance. These estimates were found to differ from nominated best estimates by 50% or more in 43% of cases, in turn leading to under or over-estimation of samplesize in the design of a notional monitoring program. The twostudies, from the same general area, using the same samplingmethods and spanning a similar time scale, gave estimatesvarying by more than an order of magnitude for 25% of taxa.We suggest that pilot studies for ecological monitoring programsof marine infauna should include at least two sampling times.     

Aerosol Scavenging: Model Application and Sensitivity Analysis in the Indian Context

Sulfate aerosols have been found to bethe major contributors to precipitation acidity. Thus,in view of the long-term ecological repercussions theyhave on aquatic ecosystems and their acidity-potential,the present analysis focuses on a case study applicationof the layer-averaged aerosol-scavenging model (Okita et al., 1996) for predicting values of the wet scavengingcoefficient and sulfate concentrations in precipitationsamples on the basis of the information available forsome selected Indian cities. Through sensitivityanalysis (Pandey et al., 1997) the scavengingcoefficient has been found to be very strongly dependenton precipitation intensity. Comparison of modelpredictions has been done with the measured values forDelhi, Mumbai, Calcutta and Chennai in India.     

Examples of the role of analytical chemistry in environmental risk management research

Analytical chemistry is an important tier of environmental protection and has been traditionally linked to compliance and/or exposure monitoring activities for environmental contaminants. The adoption of the risk management paradigm has led to special challenges for analytical chemistry applied to environmental risk analysis. Namely, methods developed for regulated contaminants may not be appropriate and/or applicable to risk management scenarios. This paper contains examples of analytical chemistry applied to risk management challenges broken down by the analytical approach and analyte for some selected work in our laboratory. Specific techniques discussed include stable association complex electrospray mass spectrometry (cESI-MS), gas chromatography-mass spectrometry (GC-MS), split-flow thin cell (SPLITT) fractionation and matrix-assisted laser desorption time of flight mass spectrometry (MALDI-ToF-MS). Specific analytes include haloacetic acids (HAA9), perchlorate, bromate, triazine degradation products, metal-contaminated colloids and Cryptosporidium parvum oocysts.     

Response of the Ascorbate-Peroxidase of Selenastrum capricornutum to Copper and Lead in Stormwaters

The green alga Selenastrum capricornutum expresses a uniqueascorbate peroxidase, that responds to copper and lead. Attemptswere made to test if this peroxidase could be used to monitor thelevels of copper and lead in natural waters. When S.capricornutum was exposed to a stormwater sample, the specificactivity of the peroxidase in the cell extract was commensuratewith the combined copper and lead contents in the sample. Theperoxidase responses were also correlated with the 96 hr biomasstoxicity assay of S. capricornutum. However, unlike thebiomass toxicity assay, the peroxidase activity was not affectedby the anions in the samples. The use of this peroxidase can beused as a marker for testing heavy metal toxicity in the water.     

Luminescence spectroscopy as a screening tool for the potential carcinogenicity of asphalt fumes

A subset of polycyclic aromatic compounds (PACs), which contain 4-6 annulated rings, has been documented as the source of carcinogenicity in animal skin painting studies of petroleum products and asphalt fumes (M. L. Machado, P. W. Beatty, J. C. Fetzer, A. H. Glickman and E. L. McGinnis, Fundam. Appl. Toxicol., 1993, 21, 492; T. A. Roy, S. W. Johnson, G. R. Blackburn and C. R. Mackerer, Fundam. Appl. Toxicol., 1988, 10, 466). Because of the chemical complexity of these materials, it has been difficult to identify the specific compounds within this broad range of PACs responsible for their carcinogenicity. An alternative approach using luminescence spectroscopy was taken in this study to quantify, without identification, a subset of these compounds that appears to cause cancer. The fluorescence response at a specific wavelength pair was obtained for 39 laboratory asphalt fume condensates from animal skin painting studies, yielding a linear correlation coefficient of R2 = 0.96 between the fluorescence response in these materials and the carcinogenicity found in animal studies. In the absence of other asphalt fume condensates from animal studies, 17 petroleum oils were also evaluated using this method and compared with the available animal skin painting data. The details of the method include a clean-up step that removes the highly polar compounds and spectral subtraction of two- and three-ring PAC interference, both of which add to the fluorescence response, yet were not found to contribute to a carcinogenic response from skin painting studies. Full scan fluorescence plots also produce a fingerprint which can be used to assess contamination, such as coal tar products or mixtures of materials, that are not defined as asphalt, yet may be present in the working environment.     

Influence of environmental parameters on the accuracy of nitrogen dioxide passive diffusion tubes for ambient measurement

Two studies at three sites in the UK provided confirmation that systematic positive bias in NO2 diffusion tube measurement occurred because of changes to "within-tube" chemistry, rather than eddy diffusion at the mouth of the tube. In the first study in Cambridge, UK, sampler overestimation for 1 and 2 week exposures was compared to corresponding time-averaged monitor measurements (NO-NO2-NOx, O3) and weather variables. Noninearity between sampler and monitor NO2 measurements was interpreted in terms of spatial and temporal variations in relative and absolute availability of NO, NO2 and O3 at the site. A maximum overestimation occurred for an exposure mean NO2/NOx approximately 0.5. The separate contributions of reduced NO2 photolysis and eddy diffusion were compared in Study II using samplers of two materials, acrylic and quartz, and of different lengths (40, 55, 71 and 120 mm) at three sites: Norwich background, Cambridge intermediate, London kerbside. For compared sites, NO2 measured by acrylic samplers was significantly higher than for equivalent quartz samplers. For quartz samplers [NO2]mean was only just above the monitor at Norwich and London; sampler/monitor NO2 = 1.04 (P = 0.59) and 1.01(P = 0.76), respectively. For acrylic samplers the order of [NO2]mean was 40 mm > 120 mm > 71 mm > or = 55 mm. Excepting 40 mm samplers, this accords with a chemical bias where co-diffusing NO and 03 molecules in longer tubes have more time to react to form excess NO2. Bias in 40 mm samplers is discussed. Eddy diffusion is negligible for standard samplers because [NO2]mean was equivalent for 55 mm and 71 mm acrylic samplers and close to monitor NO2 for 71 mm quartz tubes. Both studies showed that sampler accuracy was dependent on location. Significantly, overestimation was greatest (approximately 3-4 ppb) where the NO2 annual mean was approximately 20 ppb, close to the UK and EU air quality standard of 21 ppb.     

Formation and Emission of N2O and CH4 from Compost Heaps of Organic Household Waster

Composting can be a source of N₂O andCH₄ production. In this investigation, differentcompost heaps of organic household waste weremonitored with the focus on potential formation ofCH₄ and N₂O in the heaps and emission ofthese gases from the heaps. The studied compost heapshad different compost ages, turning intervals andcompost sizes. The analysed compost gases containedbetween 1–3421 L of N₂O-N L^-1 and 0–470 mL of CH₄ L^-1. The emission rates ofN₂O and CH₄ from the compost heaps werebetween 1–1464 mg N₂O m^-2 day^-1 and0–119 000 mg CH₄ m^-2 day^-1. These verylarge differences in compost gas composition andemission indicate the importance of compostmanagement. The results also give an understanding ofwhere in the composting process an increasing emissionof N₂O and CH₄ can occur.     

Lead in Soil and Sediment in Iqaluit, Nunavut, Canada, and Links with Human Health

This study determined the spatial distribution of soiland of sediment-associated lead in Iqaluit, Nunavut.Samples were collected from the following areas:outside the built-up area of the town to reflectbackground concentrations; known or potential pointsources of lead, such as the Upper Base, the SylviaGrinnell Dump and the Metal Dump (North 40); andresidential and commercial areas of Iqaluit and Apex,a satellite community. In the laboratory, the <63 m sample fraction was analyzed for total lead andbioavailable lead, estimated by non-residual acidextractable lead content. The research findings revealthat elevated levels of bioavailable lead are presentin the study area. Total lead concentrations generallydo not exceed environmental guidelines. However, leadconcentrations in the Sylvia Grinnell Dump, and Apexand Iqaluit grid areas exceed health-based guidelines.The research concludes that there is not a serioushealth hazard posed by lead levels in the soil andsediment in the study area. However, severalenvironmental (elevated lead levels, bioavailableforms of lead and bare soil surfaces) and behaviouralfactors (vigorous and unsupervised play outside) maycreate a risk of lead exposure.