The present study evaluated the performance and variability in acute toxicity tests with glochidia and newly transformed juvenile mussels using the standard methods outlined in American Society for Testing and Materials (ASTM). Multiple 48-h toxicity tests with glochidia and 96-h tests with juvenile mussels were conducted within a single laboratory and among five laboratories. All tests met the test acceptability requirements (e.g., >or=90% control survival). Intralaboratory tests were conducted over two consecutive mussel-spawning seasons with mucket (Actinonaias ligamentina) or fatmucket (Lampsilis siliquoidea) using copper, ammonia, or chlorine as a toxicant. For the glochidia of both species, the variability of intralaboratory median effective concentrations (EC50s) for the three toxicants, expressed as the coefficient of variation (CV), ranged from 14 to 27% in 24-h exposures and from 13 to 36% in 48-h exposures. The intralaboratory CV of copper EC50s for juvenile fatmucket was 24% in 48-h exposures and 13% in 96-h exposures. Interlaboratory tests were conducted with fatmucket glochidia and juveniles by five laboratories using copper as a toxicant. The interlaboratory CV of copper EC50s for glochidia was 13% in 24-h exposures and 24% in 48-h exposures, and the interlaboratory CV for juveniles was 22% in 48-h exposures and 42% in 96-h exposures. The high completion success and the overall low variability in test results indicate that the test methods have acceptable precision and can be performed routinely.

Beta-methylamino-L-alanine (BMAA) is a neurotoxic amino acid that can be produced by cyanobacteria in aqueous environments. To analyze this compound by gas chromatography/mass spectrometry (GC/MS), BMAA must be derivatized to a nonpolar, volatile compound. This can be accomplished by reacting BMAA with ethyl chloroformate. While carrying out electron ionization (EI) mass spectrometric analysis on the (13)C-labeled derivative, it was discovered that the formation of an ion with a peak at m/z 245.12 is the result of [CH(3)CH(2)O.] loss from the amino groups resulting from alpha-cleavage. This differs from previous reports that attributed this peak to alpha-cleavage of the carboxylic ester portion of the BMAA derivative. This finding is important for understanding BMAA derivative mass spectrometric fragmentation patterns and ultimately to properly identifying and quantifying BMAA. Fragmentation pathways for the formation of other major peaks observed in the EI mass spectra are also proposed.

In light of current interest in better understanding the environmental impact of atmospheric particulate matter (PM), a new strategy has been employed to screen the relative toxicities of ambient and source aerosols. Short-term and acute aquatic bioassays using Ceriodaphnia dubia and a green alga (Selenastrum capricornutum) as test organisms have been in use for many years in the regulation of wastewater effluents. These tests have been employed in the present study to compare the toxicity of water extracts of atmospheric particulate matter and dichloromethane (DCM) extracts that have been transferred to dimethyl sulfoxide and diluted in water. Atmospheric PM was collected at four sites located near the south shore of Lake Michigan and one site in Michigan's Upper Peninsula at discrete events during three seasons. Parallel chemical analyses of the two extracts directly assessed the relation between the chemical composition and the toxicity of the extract. Inductively coupled plasma analysis of the metals in the water extract and gas chromatography-mass spectroscopy of the organics in the DCM extract showed a relationship between high toxicity and high water-soluble copper concentration and high secondary organic aerosol tracers in the extracted aerosol. Although previous fractionation studies have not looked at water-soluble copper, significant toxicity has been measured in the semipolar and polar organic fractions of ambient aerosols and diesel exhaust particles, which are the fractions in which secondary organic aerosol components would be expected. For the water extracts, the summer samples were consistently more toxic than the autumn or spring samples. There was not a seasonal pattern for the toxicity of the DCM extracts; however, spatial differences were apparent. The toxicity end points of select samples from one site qualitatively correlate with the high polycyclic aromatic hydrocarbon concentrations. Additionally, high toxicity in the July DCM extracts from another site may be tied to the presence of the insecticide carbaril. The seasonal and spatial variations captured in the toxicity results in this study tend to qualitatively correlate with trace organic components and metals and not bulk particulate matter composition.