Quantitative Structure-Activity Relationship, Ontology-Based Model of the Antioxidant and Cell Protective Activity of Peat Humic Acids.

Peat humic acids are well known for their wide range of biological effects which can be attributed to the complex chemical structure of naturally occurring humic substances. One of the promising tools is an ontology-based quantitative analysis of the relationship between physical and chemical parameters describing a chemical structure of peat humic acids and their biological activity. This article demonstrates the feasibility of such an approach to estimate the antioxidant and cell protective properties of the peat humic acids. The structural parameters of the peat humic acids were studied by electronic, fluorescence, infrared, 13C-NMR spectroscopy, titrimetric analysis, elemental C,H,N, and O- analysis, and gel chromatography. Antioxidant and antiradical activities were assessed by physicochemical methods of analysis: electronic paramagnetic resonance, cathodic voltammetry, ABTS•+ scavenging, assay of DPPH radical-scavenging activity, assay of superoxide radical-scavenging activity, iron chelating activity, and scavenging of hydroxyl radicals. Cytoprotective activity was evaluated by the neutral red-based cytotoxicity test in 3T3-L1 cell culture in a wide range of concentrations. Assessment of intracellular ROS production was carried out using a 2,7-dichlorodihydrofluoresceindiacetate (DCFDA) fluorescent probe. Intracellular ROS production was induced using two common prooxidants (tert-butyl hydroperoxide, Fe2+ ions). We suggested an ontology-based model for the antioxidant and cytoprotective activity of humic acids based on experimental data and numerical models. This model establishes the way to further research on the biological effects of humic acids and provides a useful tool for numerical simulation of these effects. Remarkable antioxidant and cell protective activity of humic acids makes them a promising natural source of new pharmaceutical substances that feature a wide range of biological effects.

New artificial network model to estimate biological activity of peat humic acids.

PURPOSE: This article focuses on new method to estimate biological activity of peat humic acids (HAs) using artificial neural network (ANN) to process spectroscopic measurements in infrared and visible ranges. Conventional approaches generally rely on biological models and direct detection of chemical substances related to bioactivity. These methods proved to be accurate and reliable, but at the expense of speed and simplicity. MATERIALS AND METHODS: Recently, a conception of quantitative structure-activity relationship (QSAR) has been introduced and successfully implemented to predict effects of HAs on toxicity of polycyclic aromatic hydrocarbons. Our research stems from this conception, but employs multilayer perceptron (MLP) model to improve overall performance. The developed MLP model allowed us to estimate biological activity of the complete vertical peat cores collected from oligotrophic peat bog, located in southern taiga zone of West Siberia (north-eastern spurs of the Great Vasyugan Mire, 56°58' N 82о36' E). In total, 42 samples taken from the cores were collected. The protocol included spectroscopy (in infrared and visible ranges) and biological model with peritoneal activated macrophages as a reference method to directly measure biological activity of HAs. RESULTS: and discussion. Numerical experiments confirmed consistency of the measured and estimated bioactivity, coefficient of determination R2 = 0.97. These experiments also showed that the MLP model significantly outperforms conventional linear multiple regression models, mainly due to essential nonlinearity of structure-activity relationships. CONCLUSIONS: Our research demonstrates that biological activity of HAs extracted from peat samples can be estimated using an artificial neural network model trained on infrared and visible spectra.

Physicochemical Characterization and Antioxidant Activity of Humic Acids Isolated from Peat of Various Origins.

Although humic acids (HAs) from peat exhibit various therapeutic properties, there is little information available concerning their physicochemical and antioxidant properties. To address this issue, nine different types of peat, including oligotrophic, mesotrophic, and minerotrophic peat samples, were used for isolation of HA fractions by basic (HAb) and pyrophosphate (HAp) extractions. Physical parameters of the HAs were analyzed by UV-Vis, fluorescent, infrared (IR), and electron paramagnetic resonance (EPR) spectroscopy. Average Mr of the fractions ranged from 17.2 to 39.7 kDa, while their humification index (HIX) varied from 0.49 to 1.21. HAp fractions had a higher content of aromatic structures compared to HAb fractions. Moreover, HAp fractions had a significantly higher content of phenolic OH groups (3.6 ± 0.5 mmol/g) versus HAb (3.1 ± 0.5 mmol/g). All HA fractions exhibited antioxidant activity in radical scavenging and electrochemical assays, and their EPR signal had a single line with g = 2.0035, which is consistent with semiquinone type radicals. Furthermore, the HIX was found to be important in determining the number of semiquinone-type free radicals in the HA structures. Overall, these data provide a molecular basis to explain at least part of the beneficial therapeutic properties of peat-derived HAs.