Stability of Se species in plant extracts rich in phenolic substances
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Since there is growing awareness of the strong dependence of the antioxidative function of selenium (Se) upon its chemical form, the stability of Se species during sample preparation is an important factor in obtaining qualitative and quantitative results. Many plant samples are rich in phenolic compounds (antioxidants), but data about their effect on specific Se species in extracts of plant samples are scarce. Therefore, the aim of this study was to investigate the effect of the most common phenolic substances in plant parts, namely tannin and the flavonoid rutin, on the concentration and/or transformation of several Se species (SeMet, SeCys2, SeMeSeCys, Se(VI) and Se(IV)) during sample preparation (24 h incubation at 37 °C) and storage (4 days at 4 °C). Moreover, the effect observed was then studied in a real sample, buckwheat, because this plant is known as a rich source of phenolics, especially tannin and rutin. Se speciation was carried out by on-line coupling of ion-exchange HPLC-ICP-MS after water and enzymatic (protease) hydrolysis. The results showed that the ratio between the two antioxidants has an important role. When the antioxidants were present together, the response for Se(IV) was observed to start to decrease only at a ratio of rutin to tannin of 1:100 (w/w), indicating the ratio between antioxidants in buckwheat seeds. After water extraction, only 40% and after enzymatic extraction 80% of Se(IV) remained, but no other Se compound was detected with the system used. Furthermore, the extracts were not stable during storage at 4 °C. Signals for other Se species were stable. The results obtained for buckwheat seeds showed a decrease in Se(IV) response during sample preparation and storage, comparable to the one obtained with the experiments performed in vitro. However, Se species in extracts of other buckwheat parts (leaves, stems and sprouts) were stable. These results indicate that reactions in the extraction process and during storage may affect Se speciation and may result in misidentifications and inaccurate values.
KeywordsSe species Phenolic substances Stability Buckwheat HPLC-ICP-MS Trace elements Quality assurance/control Biological samples
This work was financially supported by the Slovenian Research Agency through the programme P1-0143, contract 1000-05-310030 and project J7-9805. The authors would like to thank Prof. Ivan Kreft for donating the buckwheat seeds and Miss Maja Vogričič for milling them.
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