Part review, part perspective, this article examines the applications and potential of in-vivo Nuclear Magnetic Resonance (NMR) for understanding environmental toxicity. In-vivo NMR can be applied in high field NMR spectrometers using either magic angle spinning based approaches, or flow systems. Solution-state NMR in combination with a flow system provides a low stress approach to monitor dissolved metabolites, while magic angle spinning NMR allows the detection of all components (solutions, gels and solids), albeit with additional stress caused by the rapid sample spinning. With in-vivo NMR it is possible to use the same organisms for control and exposure studies (controls are the same organisms prior to exposure inside the NMR). As such individual variability can be reduced while continual data collection over time provides the temporal resolution required to discern complex interconnected response pathways. When multidimensional NMR is combined with isotopic labelling, a wide range of metabolites can be identified in-vivo providing a unique window into the living metabolome that is highly complementary to more traditional metabolomics studies employing extracts, tissues, or biofluids.
A dominant source of light-absorbing aerosol particles, brown carbon (BrC), to the atmosphere is smoke from biomass burning. Aqueous aging of biomass burning organic aerosol can increase BrC absorbance, which may extend its atmospheric lifetime in aerosol particles, cloud droplets, and fog droplets. This study investigates the aqueous aging of biomass burning BrC and the connection between absorbance and chemical composition. The water-soluble component of laboratory-generated wood smoke BrC was analyzed using aerosol-chemical ionization mass spectrometry, liquid chromatography–mass spectrometry (electrospray ionization), UV–vis spectroscopy, and NMR spectroscopy as it was exposed to UV-B light and OH oxidation to simulate photo-oxidation in the atmosphere. During UV-B light exposure, absorbance at 400 nm increased by greater than a factor of 2 and remained high for the 6 h exposure period. A similar increase in absorbance was observed during OH oxidation, up to an OH exposure of 4 × 10–10 M·s. At a cloud water OH concentration of 1 × 10–14 M, this OH exposure corresponds to ∼11 h of aqueous OH oxidation. Further OH oxidation led to a net loss of absorbance after an OH exposure of 1.5 × 10–9 M·s (∼42 h of aqueous OH oxidation). The increase in absorbance in both cases was linked to the formation of aromatic dimer compounds and functionalized products only during OH oxidation. The loss of absorbance with extended OH oxidation correlated with a loss of aromatic compounds and breakdown to smaller molecules. These results show that aqueous aging of the biomass burning material through photo-oxidation primarily increases the absorbance of BrC and may result in longer-lived BrC in the atmosphere.
Nuclear Magnetic Resonance (NMR) spectroscopy is a non-invasive analytical technique which allows for the study of intact samples. Comprehensive Multiphase NMR (CMP-NMR) combines techniques and hardware from solution state and solid state NMR to allow for the holistic analysis of all phases (i.e. solutions, gels and solids) in unaltered samples. This study is the first to apply CMP-NMR to deceased, intact organisms and uses 13 C enriched Daphnia magna (water fleas) as an example. D. magna are commonly used model organisms for environmental toxicology studies. As primary consumers, they are responsible for the transfer of nutrients across trophic levels, and a decline in their population can potentially impact the entire freshwater aquatic ecosystem. Though in vivo research is the ultimate tool to understand an organism’s most biologically relevant state, studies are limited by conditions (i.e. oxygen requirements, limited experiment time and reduced spinning speed) required to keep the organisms alive, which can negatively impact the quality of the data collected. In comparison, ex vivo CMP-NMR is beneficial in that; organisms do not need oxygen (eliminating air holes in rotor caps and subsequent evaporation); samples can be spun faster, leading to improved spectral resolution; more biomass per sample can be analyzed; and experiments can be run for longer. In turn, higher quality ex vivo NMR, can provide more comprehensive NMR assignments, which in many cases could be transferred to better understand less resolved in vivo signals. This manuscript is divided into three sections: 1) multiphase spectral editing techniques, 2) detailed metabolic assignments of 2D NMR of 13 C enriched D. magna and 3) multiphase biological changes over different life stages, ages and generations of D. magna . In summary, ex vivo CMP-NMR proves to be a very powerful approach to study whole organisms in a comprehensive manner and should provide very complementary information to in vivo based research.
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