High-resolution 1H nuclear magnetic resonance (NMR) analysis was employed to molecularly screen the lipid, lipid oxidation product (LOP), and antioxidant compositions of four natural (unrefined) cod liver oil (CLO) products. Products 1–3 were non-fermented CLOs, whilst Product 4 was isolated from pre-fermented cod livers. Supporting analytical data that were acquired included biogenic amine, flavanone, tannin, phenolic antioxidant, α-tocopherol, and oxygen radical absorbance capacity (ORAC) determinations by recommended HPLC, LC/MS/MS, or spectrophotometric methods. SDS-PAGE, HPLC, and 1H NMR analyses investigated and determined collagenous antioxidants and their molecular mass ranges. 1H NMR analysis of aldehydic LOPs was employed to explore the susceptibilities/resistivities of each CLO product to peroxidation that is induced by thermal stressing episodes (TSEs) at 180°C, or following prolonged (42 day) storage episodes at 4 and 23 °C. Product 4 displayed extremely high ORAC values, which were much greater than those of Products 1–3, and that were predominantly ascribable to significant levels of peroxidation-blocking and/or aldehyde-consuming collagenous polypeptides/peptides and ammoniacal agents therein. Significantly lower levels of toxic aldehydes were generated in the pre-fermented Product 4 during exposure to TSEs, or the above long-term storage episodes. These results confirmed the enhanced peroxidative resistivity of a fermented, antioxidant-fortified natural CLO product over those of non-fermented unrefined products. Product 4: Green Pasture Blue Ice™ Fermented Cod Liver Oil.
Soybean oil is the second most exported oil from the United States and South America, and is widely marketed as a cooking oil product containing numerous health benefits for human consumers. However, culinary oils with high polyunsaturated fatty acid (PUFA) contents, are known to produce high quantities of lipid oxidation products (LOPs), including toxic aldehydes upon exposure to high-temperature frying episodes. Previous studies have demonstrated causal links between aldehyde ingestion and inhalation with deleterious health perturbations, including mutagenic and carcinogenic effects, along with cardiovascular and teratogenic actions. In this study, aldehydic LOPs were detected and quantified in commercially available samples of soybean, avocado, corn and extra-virgin olive oil products before and after their exposure to laboratory-simulated laboratory frying episodes (LSSFEs) using high-resolution 1H nuclear magnetic resonance (NMR) analysis. Results acquired demonstrated that PUFA-rich soybean and corn oils gave rise to the highest concentrations of oil aldehydes from the thermo-oxidation of unsaturated fatty acids, whereas monounsaturated fatty acid (MUFA)-laden avocado and olive oils were much more resistant to this peroxidation process, as expected. Multivariate chemometrics analyses provided evidence that an orthogonal component pattern of aldehydic LOPs featuring low-molecular-mass n-alkanals such as propanal, and 4-oxo-alkanals, arises from thermo-oxidation of the ω-3 fatty acid (FA) linolenic acid (present in soybean oils at levels of ca. 7% (w/w)), was able to at least partially distinguish this oil from corresponding samples of thermally-stressed corn oil. Despite having a similar total PUFA level, corn oil has only a negligible ω-3 FA content, and therefore generated significantly lower levels of these two aldehyde classes. In view of the adverse health effects associated with dietary LOP ingestion, alternative methodologies for the incorporation of soybean oils within high-temperature frying practices are proposed.
Implementations of high-field nuclear magnetic resonance (NMR) facilities into metabolomics studies are unfortunately restricted by their large dimensions, high costings, and specialist technical staff requirements. Therefore, here the application and practical advantages offered by low-field (60 MHz), compact NMR spectrometers for probing the metabolic profiles of human saliva was explored, as was their value in salivary metabolomics studies. Saliva samples were collected from cigarette smoking (n = 11) and non-smoking (n = 31) human participants. 1H NMR spectra were acquired on both low-field (60 MHz) and medium-field (400 MHz) spectrometers. Metabolomics analyses were employed to evaluate the consistencies of salivary metabolite levels determined, and their abilities to distinguish between smokers and non-smokers. Low-field 1H NMR analysis detected up to 15, albeit permitted the reliable quantification of 5, potentially key diagnostic biomolecules simultaneously (LLOQ values 250–400 μmol/L), although these were limited to those with the most prominent resonances. Such low-field profiles were also found to be suitable for salivary metabolomics investigations, which confirmed the successful discrimination between smoking and non-smoking participant sample donors. Differences observed between these groups were largely ascribable to upregulated salivary levels of methanol, and its metabolite formate, in the smoking group, but higher smoking-mediated concentrations of acetate, propionate and glycine may arise from a diminished salivary flow-rate in these participants. In conclusion, determination of salivary biomolecules using low-field, benchtop 1H NMR analysis techniques were found to be valuable for bioanalytical and metabolomics investigations. Future perspectives for the applications of this non-stationary NMR technique, for example for the on-site ‘point-of-care’ testing of saliva samples for diagnostic oral disease screening purposes at dental surgeries and community pharmacies, are considered.
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