The Toolbox of Methods for Multidirectional Characterization of Dietary Nucleic Acids; Verification for Raw and Processed Food Products

Abstract

AbstractCurrently, the nutritional value of food is associated mainly with components such as proteins, carbohydrates, and lipids. However, another important macromolecules present in many foods are dietary nucleic acids (dietNA), i.e., DNA as well as both coding and non-coding RNAs. In the context of food chemistry and nutrition, dietNA are nowadays vastly neglected. In consequence, there are no dedicated methodologies to characterize dietNA. In this study, using raw or processed meat and plant products as model foodstuffs, we developed a toolbox of methods borrowed from other fields (histology, toxicology, molecular biology) that enable the initial characterization of dietNA as a necessary step on the way to systematic evaluation of their nutritional role. The proposed set of methods embraces (i) paraffin embedding of food samples and their staining to visualize the distribution and variety of dietNA in situ; (ii) comet assay to assess integrity of nuclear DNA with possible detection of DNA damage; (iii) dietNA isolation with and without RNAse digestion to determine the content of both DNA and RNA; (iv) electrophoretic separation of isolates to profile dietNA fragments. Such a combined methodological approach revealed clear differences between dietNA derived from raw and processed food products. We believe that the presented set of methods will encourage the broader research on dietNA to understand their role as a nutritionally relevant food component.