The ABTS and DPPH methods are among the most popular assays of antioxidant activity determination. Attempts to adapt them to different analytes and the search for the highest values of antioxidant activity has resulted in a large variety of assay conditions to be presented in the literature, including the way the measurement is made. This makes it difficult to relate the results to real oxidation systems, and often makes it impossible to compare them. Such a comparison is limited in advance by the use of stable radicals that do not exist in nature and that react differently from those generated in food or in vivo. Therefore, it is important to introduce measures aimed at standardizing the conditions of the activity assay, including reaction time and several reaction environments suitable for testing different groups of compounds. In this study, we used natural antioxidants of various structures: phenolic acids, flavonoids, peptides and corresponding amino acids, ascorbic acid and α-tocopherol, and also synthetic analogues of selected compounds. The curves of dependence of the measured absorbance on the concentration of antioxidants were described, the ranges of linearity were determined, and the value of the error made when reading in various ranges of dependencies was estimated. We also determined and compared the activity values using two popular methods (IC50 and TEAC), taking into account different environments and reaction times. Based on the collected data, recommendations were formulated regarding the reaction conditions adapted to the studies of individual groups of antioxidants, and unified reaction times were proposed. Taking into account the state before reaching the equilibrium of antioxidants reacting in a complex manner, this approach may introduce a simplified reference to the competing reaction that occurs in reality.
The aim of this research was to establish the effect of mild roasting on coffee beans contamination level by polycyclic aromatic hydrocarbons (PAHs).
The materials investigated were green Arabica and Robusta coffee beans imported from different countries, as well as those already roasted.
The experiment was carried out in a coffee-roasting plant, with the use of an electric coffee roaster, at the temperature of 125–135 °C for 25–26 min. PAHs analysis was conducted by means of high-performance liquid chromatography with fluorescence and diode array detectors (HPLC-FLD/DAD). Results had been verified by means of gas chromatography with mass spectrometry.
Contamination level for 19 PAHs, 15 of which were heavy PAHs included on the list of European Union Scientific Committee in Food, varied from 4.29 to 16.17 µg/kg in roasted coffee beans, whereas in green coffee beans varied from 8.66 to 76.63 µg/kg. The results of statistical analysis showed that the contamination level in roasted coffee beans was significantly lower than that in green beans. The applied parameters of roasting did not lead to the occurrence of conditions in which PAHs, especially heavy ones, would possibly be formed. On the contrary, the roasting process itself had significantly reduced the PAHs content in the final product. The reason for this phenomenon was relatively high volatility of light PAHs.
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