Neoformation of antioxidants in glycation model systems treated under subcritical water extraction conditions

In this work, the possibility of finding newly formed Maillard reaction products produced as a result of the subcritical water extraction (SWE) conditions is explored. Simplified powdered glycation model systems were prepared mixing amino acid (Lys, Arg or Ala) and glucose in a molar ratio 1:4. Samples constituted by glucose or amino acids alone were also prepared as controls. SWE was carried out at room temperature, 100 °C and 200 °C and 100 bar of pressure for 20 min. Different assays were performed in order to determine the extent of glycation by analyzing the decrease of free amino groups and/or the formation of Maillard reaction products (early, advanced and end products). Namely, formation of early colorless Maillard reaction products, Amadori compounds, was detected by ESI-MS; advanced glycation end products (AGEs) were detected by measuring the fluorescence intensity (λ_ex = 360 nm, λ_em = 460 nm) while end brown products were detected by reading the absorbance at 360 and 420 nm. Besides, the antioxidant capacity of the aqueous extracts was determined by using ABTS and ORAC_FL assays. Results obtained indicated the occurrence of the Maillard reaction under our specific extraction conditions. Early, advanced and end products were detected in the samples. Caramelization of sugar also occurred. As expected, the extent of the non-enzymatic browning depended on the intensity of the thermal treatment. Additionally, data on antioxidant activity suggested the formation of neoantioxidants. These compounds were predominantly formed at 200 °C. In conclusion, this report demonstrates the formation of antioxidant compounds in simplified glycation model systems under SWE conditions.