The metabolic response of the earthworm Eisenia fetida to two pesticides, dichlorodiphenyltrichloroethane (DDT) and endosulfan, was characterized in contact tests using proton nuclear magnetic resonance ( 1 H NMR) and principal component analysis (PCA). PCA loading plots suggested that maltose, leucine and alanine were important metabolites contributing to the differences in dosed and control earthworms for both compounds at doses of 0.5, 1.0 and 2.0 lg/cm 2 . Gas chromatography/ mass spectrometry (GC/MS) was used to quantify the metabolites identified in E. fetida and determine if the changes in maltose, leucine and alanine following exposure to DDT and endosulfan (at 0.5 and 1.0 lg/cm 2 ) were reproducible and greater than the natural variability. Quantification by GC/MS suggested that maltose was not a reliable biomarker since it both increased and decreased in earthworms exposed to DDT and increased by just 3% with exposure to endosulfan. Leucine was not stable with the GC/MS derivitization method used in this study and could not be confirmed as a reliable biomarker. However, alanine consistently increased for both DDT and endosulfan exposed E. fetida. Alanine showed considerable variability in control earthworms (±41.6%), yet the variability in alanine to glycine ratios was just ±10.5%. Increases in the alanine to glycine ratio were statistically significant at the P = 0.05 level for the 1.0 lg/cm 2 DDT dose and both the 0.5 and 1.0 lg/cm 2 endosulfan doses, suggesting that deviations from the normal homeostatic ratio of 1.5 for alanine to glycine is a potential biomarker of DDT and endosulfan exposure warranting further study.
Flow injection mass spectrometry and proton nuclear magnetic resonance spectrometry, two metabolic fingerprinting methods, and DNA sequencing were used to identify and authenticate Actaea species. Initially, samples of Actaea racemosa from a single source were distinguished from other Actaea species based on principal component analysis and soft independent modeling of class analogies of flow injection mass spectrometry and proton nuclear magnetic resonance spectrometry metabolic fingerprints. The chemometric results for flow injection mass spectrometry and proton nuclear magnetic resonance spectrometry agreed well and showed similar agreement throughout the study. DNA sequencing using DNA sequence data from two independent gene regions confirmed the metabolic fingerprinting results. Differences were observed between A. racemosa samples from four different sources, although the variance within species was still significantly less than the variance between species. A model based on the combined A. racemosa samples from the four sources consistently permitted distinction between species. Additionally, the combined A. racemosa samples were distinguishable from commercial root samples and from commercial supplements in tablet, capsule, or liquid form. DNA sequencing verified the lack of authenticity of the commercial roots but was unsuccessful in characterizing many of the supplements due to the lack of available DNA.
Environmental contextEnvironmental metabolomics is an emerging field that examines the metabolic changes in organisms in response to potential environmental stressors. In this study, nuclear magnetic resonance spectroscopy is used to investigate earthworm metabolic responses to sub-lethal exposure of environmentally persistent pesticides. The study identifies two toxic modes of action elicited by the pesticides, and highlights the potential of metabolomics for the chemical assessment of persistent environmental contaminants. Abstract1-D and 2-D nuclear magnetic resonance (NMR) spectroscopy is used to examine the metabolic response of the earthworm (Eisenia fetida) after contact test exposure to an organofluorine pesticide, trifluralin, and an organochlorine pesticide, endosulfan. Three sub-lethal concentrations were used for each pesticide (0.1, 0.5 and 1.0 mg cm–2 for trifluralin and 0.5, 1.0 and 2.0 μg cm–2 for endosulfan). Principal component analysis of the trifluralin and endosulfan NMR datasets showed separation between the unexposed and the exposed earthworm groups. Alanine, glycine, maltose and ATP were significant in the highest concentration (1.0 mg cm–2) for trifluralin-exposed earthworms and may result from a non-polar narcosis toxic mode of action (MOA). Leucine, phenylalanine, tryptophan, lysine, glutamate, valine, glycine, isoleucine, methionine, glutamine, alanine, maltose, glucose, meibiose, malate, fumarate and ATP were detected as significant for the two highest concentrations (1.0 and 2.0 μg cm–2) for endosulfan-exposed earthworms and a neurotoxic MOA is postulated. This study highlights the use of 1-D and 2-D metabolomics for understanding the biochemical response of environmental contaminants to model organisms such as earthworms.
Acer truncatum is an important ornamental, edible, and medicinal plant resource in China. Previous phytochemical research has focused on the leaf (AL) due to its long history as a tea for health. Other parts such as the branch (ABr), bark (ABa), fruit (AF), and root (AR) have drawn little attention regarding their metabolites and bioactivities. The strategy of an in-house chemical library combined with Progenesis QI informatics platform was applied to characterize the metabolites. A total of 98 compounds were characterized or tentatively identified, including 63 compounds reported from this species for the first time. Principal component analysis showed the close clustering of ABr, ABa, and AR, indicating that they share similar chemical components, while AL and AF clustered more distantly. By multiple orthogonal partial least-squares discriminant analyses (OPLS-DA), 52 compounds were identified as potential marker compounds differentiating these different plant parts. The variable influence on projection score from OPLS-DA revealed that catechin, procyanidins B2 or B3, and procyanidins C1 or C2 are the significant metabolites in ABa extracts, which likely contribute to its antioxidant and cytotoxic activities.
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