Long chain fatty acids analysis of intertidal biofilm by direct injection liquid chromatography time of flight mass spectrometry.

The critical importance of mono- and polyunsaturated fatty acids (FAs) in a variety of biological functions, including animal nutrition and as an environmental stress monitor, is well recognized. However, while methods exist for monitoring of fatty acids, few are specific either to the profile of a microphytobenthos matrix or practical in application to multiple, diverse intertidal biofilm sample sets. In the current study, a sensitive liquid chromatography (LC) quadrupole time of flight mass spectrometry (QTOF) method was developed for the quantitative analysis of 31 FAs specific to intertidal biofilm, a thin mucilaginous layer of microalgae, bacteria, and other organisms on the surface of coastal mudflats, which provide a rich source of FAs for migratory birds. Preliminary screening of diverse biofilm samples collected from shorebird feeding grounds highlighted eight saturated (SFA), seven monounsaturated (MUFA), and sixteen polyunsaturated FAs (PUFA) that were selected for analysis. Improved method detection limits in the range 0.3-2.6 ngmL-1 were achieved, excepting for stearic acid at 10.6 ngmL-1. These excellent results were obtained without use of complex sample extraction and clean-up procedures undertaken by other published methods. An alkaline matrix of dilute aqueous ammonium hydroxide with methanol was shown to be selective for extraction and stability of the more hydrophilic fatty acid components. The direct injection method showed excellent precision and accuracy both during validation and application to hundreds of real-world intertidal biofilm samples from the Fraser River estuary (British Columbia, Canada) and other areas of the region frequented by shoreline birds.

Trace analysis of resin acids in surface waters by direct injection liquid chromatography time of flight mass spectrometry and triple quadrupole mass spectrometry.

A rapid and sensitive liquid chromatography (LC) quadrupole time of flight (QTOF) method has been developed for the determination of resin acid concentrations in aqueous pulp and paper effluent related samples. Calibration R2 of ≥0.995 for twelve resin acids, namely dehydroabietic, 8(14)-abietenic, dihydroisopimaric, levopimaric, neoabietic, pimaric, sandaracopimaric, abietic, isopimaric, palustric, chlorodehydroabietic, and dichlorodehydroabietic acids, was demonstrated in the range 1 µgL-1 to 40 µgL-1. An improved lower limit of quantitation was achieved without use of complex sample extraction and clean-up procedures undertaken by other published methods. Excellent precision and accuracy results were achieved for dehydroabietic, chlorodehydroabietic, dichlorodehydroabietic, isopimaric (integrated inclusive of all C20H30O2 resin acids), dihydroisopimaric and 8(14)-abietenic resin acids, with t-99 percentile detection limits spanning the range 0.05 to 0.07 µgL-1. While measurement for the C20H30O2 resin acids by isopimaric equivalence is considered semi-quantitative and could be an under estimate for the abietic acid component, the developed method demonstrated clear advantage over time consuming, hazardous, and unstable derivatization procedures used for gas chromatography and capillary electrophoresis. The developed LC/QToF method was successfully transferred to an LC triple quadrupole mass spectrometer for routine high throughput trace level analysis. Real world samples, including sea water and estuary water, demonstrated excellent spike recoveries by this procedure, indicating that the method is well suited to the monitoring of industrially derived resin acids in environmental surface waters. While no interferences were observed during routine sample analysis using myristic-1-13C acid and palmitic-1-13C acid internal standards, these were later substituted by myristic-d27 and palmitic-d31 acid in order to improve method robustness for environmental samples where endogenous parent fatty acids could be present.

Dioctyl Sodium Sulfosuccinate as a Potential Endocrine Disruptor of Thyroid Hormone Activity in American bullfrog, Rana (Lithobates) catesbeiana, Tadpoles.

The thyroid hormones, thyroxine (T4) and triiodothyronine (T3), are required to regulate complex developmental processes in vertebrates and are highly sensitive to endocrine-disrupting compounds. Previous studies demonstrate that dioctyl sodium sulfosuccinate (DOSS), a common constituent of pharmaceuticals, cosmetics, and food products, disrupts canonical signaling of adipocyte differentiation by binding a nuclear hormone receptor in the same superfamily as thyroid hormone (TH) receptors. The present study was designed to determine whether DOSS is capable of disrupting TH signaling using the American bullfrog, Rana (Lithobates) catesbeiana-a cosmopolitan frog species that undergoes TH-dependent metamorphosis to transition from an aquatic tadpole to a terrestrial juvenile frog. Premetamorphic R. catesbeiana tadpoles were injected with 2 pmol/g body weight T3 or 10 pmol/g body weight T4 to induce precocious metamorphosis, then exposed for 48 h to environmentally or clinically relevant DOSS concentrations (0.5, 5, and 50 mg/L). Gene expression of three classical TH-responsive targets (thra, thrb, and thibz) was measured in tadpole liver and tail fin tissue through reverse transcription quantitative polymerase chain reaction (RT-qPCR). DOSS disrupted gene expression in liver and tail fin tissue at all three concentrations tested but the patterns of expression differed by tissue, gene transcript, and TH treatment status. To our knowledge, this is the first demonstration that DOSS can alter TH signaling. Further exploration into DOSS disruption of TH signaling is warranted, because exposure may affect other TH-dependent processes, such as salmon smoltification and perinatal human development.