## 3 Results and Discussion
The same model of inhibition was exhibited by BE compounds from GCB (Figure 3B) and the mixture of GCB/WF (1:1, w/w) (Figure 3D), except for digested (DE) that seems to be non-competitive. Also, compounds released from GCB during digestion in vitro acted as competitive LOX inhibitors, whereas compounds potentially bioavailable demonstrated a competitive mode of LOX inhibition (Figure 3B). These results confirm those published previously [32]. Regardless of the extraction system, compounds contained in WF acted as non-competitive LOX inhibitors (Figure 3C). The same mode of action was found for potentially bioaccessible (DE) compounds from GCB/WF mixtures, whereas potentially bioavailable (AE) phytochemicals acted as competitive LOX inhibitors (Figure 3D). [PERSON] and [PERSON] [5] and [PERSON] et al. [33] proved that phenolic acids included in GCB act as LOX inhibitors and are responsible for the protective effect of coffee against inflammation, colon cancer and tumor promotion. As published previously, chlorogenic and ferulic acids acted as competitive inhibitors of xanthine oxidase (XO). The same mode of inhibition was exhibited by their mixture (1:1). Buffer-extractable phytochemicals from GCB acted as uncompetitive inhibitors of XO. The same mode of action was found for extracts obtained after simulated absorption, whereas potentially bioaccessible compounds acted as non-competitive XO inhibitors [20]. Consumer demand for healthy diets and convenient foods creates a huge commercial market for functional foods. Food fortification is one of the major techniques used to create functional food products with predicted pro-health activities. In this context, we proposed the new functional product--wholemeal wheat bread enriched with GCB [11]. As described previously [12], GCB addition significantly increased the LPO activity of wholemeal wheat bread. Moreover, this activity was significantly influenced by the method of extraction. The highest level of activity was found in the case of extracts obtained after simulated digestion  
Figure 3: Mode of LOX inhibition by pure ferulic and chlorogenic acids (**A**), extracts from green coffee beans (**B**), extracts from wholemeal wheat flour (**C**) and their mixture 1:1 (**D**). (potentially bioaccessible compounds, DE). As presented in Table 2, GCB addition significantly enriched wheat bread with buffer-extractable (potentially mastication-extractable) compounds able to inhibit lipid peroxidation. Their activity was positively correlated with the percentage of GBC (R\({}^{2}\) = 0.98). Simulated digestion released active compounds from all samples; however, the activity of control bread extract was significantly lower than those determined for samples from enriched breads. Unfortunately, active compounds were poorly bioavailable in vitro. Also, in this case enrichment of breads with GCB resulted in an increase in LPO activity. Use of isobolographic analysis is not possible in many food systems, thus, for analysis of interactions between active components of functional breads, the IF analysis was performed. Taking into account IF values, it may be concluded that LPO-active compounds, regardless of the type of extract, acted additively (Table 2). All breads (control and enriched with GCB) contained buffer-extractable (BE) LOX inhibitors. Addition of GCB to bread significantly improved the LOX-inhibitory activity of buffer extracts. Most importantly, BE LOX inhibitors derived from both raw materials used for bread-making acted synergistically (Table 2). The same interaction was confirmed for raw materials before baking (Figure 2D). On this basis, it can be assumed that thermal treatment does not reduce the activity and/or compounds formed during baking (e.g., Maillard reaction products (MMPs) do not interfere with the activity or even participate in it). Digestion in vitro released LOX inhibitors from all samples; however, the influence of GCB supplementation was less visible than in the case of buffer extracts.