Subcritical water extraction (SWE) is a very promising technique for obtaining bioactives (mainly antioxidants) from natural sources; even if sometimes the high operation temperatures have been suggested as responsible for thermal degradation of bioactives, the fact is that this type of extraction processes may generate new bioactive (antioxidant) compounds. The present study involved the analysis of antioxidants either naturally found in raw samples and/or those formed during extraction via Maillard reaction and other chemical events. Samples of different nature like microalgae (Chlorella vulgaris), algae (Sargassum vulgare, Porphyra spp., Cystoseira abies-marina, Sargassum muticum, Undaria pinnatifida, and Halopitys incurvus) and plants (rosemary, thyme and verbena) were studied. Amino acid availability, sugar content, fluorescence and absorbance at different wavelengths were determined to follow chemical changes due to reactions such as Maillard, caramelization and thermoxidation. Folin reaction also provided information related to total phenol content of the samples. ABTS• + , peroxyl as well as superoxide radical scavenging assays were used to measure the antioxidant capacity of the extracts. Results obtained from this study suggest that neoformed compounds derived from Maillard, caramelization and thermoxidation reactions affect the overall antioxidant capacity of water subcritical extracts depending on the nature of the sample. The brown algae U. pinnatifida was the sample in which these chemical events contributed to a higher extent to improve the antioxidant capacity (from 0.047 to 1.512 mmol/g and from 45.356 to 1522.692 μmol/g for the TEAC and ORAC FL methods, respectively) when the extraction temperature was raised from 100 to 200°C. To the best of our knowledge, this is the first work supporting the formation of neoantioxidants in natural complex matrices during subcritical water extraction.

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BACKGROUND: Aqueous pomegranate seed extract (PSE), a by-product of the pomegranate juice industry, was recently identified as a potential antiglycative ingredient. Ellagic acid was proposed as the major polyphenol responsible for the antiglycative activity as exerted in in vitro models. However, there is no information on safety aspects of this extract in biological systems before its application as ingredient. The cytotoxicity of PSE (1-100 μgmL−1) was evaluated by determining its effect on cell viability and redox status of cultured HepG2 cells. The protective effect of the PSE against oxidative stress induced by tert-butyl hydroperoxide (t-BOOH) was also investigated. RESULTS: No changes in cell integrity or intrinsic antioxidant status resulted from a direct treatment with aqueous PSE, even at high dosage. In addition, reactive oxygen species (ROS) induced by t-BOOH were reduced by 21% when cells were pretreated with 100 μg mL−1 of aqueous PSE at 180 min. The range of concentrations investigated was effective in decreasing the ROS formation but not in a dose-dependent manner.