## 3 Results and Discussion
Concentrations ranging from 1 to 25 mg/mL were tested and compared with aminoquantiidine (4 mg/mL), a compound commonly used as a positive control in antiglycation studies due to its well-documented ability to inhibit AGE formation [22]. AGE inhibition increased with higher extract concentrations, ranging from 8.3% at 1 mg/mL to 81.0% at 25 mg/mL in the BSA-Glu assay (Figure 1A). Figure 1: Effect of carob flour extracts on AGE formation in BSA–Glu (**A**) and BSA–MGO (**B**) models. Results are expressed as mean \(\pm\) SD. Different letters indicate statistically significant differences between the aqueous and methanolic extracts (\(p<0.05\)). The dashed line represents 50% inhibition. In the BSA-MGO assay, 1 mg/mL showed no significant inhibitory effect, while 25 mg/mL achieved inhibition levels close to 70% (Figure 1B). In both models, methanolic extracts exhibited significantly greater antiglycation activity than aqueous extracts, with statistically significant differences observed at 1 and 10 mg/mL in the BSA-Glu assay and at 10 mg/mL in the BSA-MGO assay. None of the extracts matched the inhibitory potency of aminoguanidine, which showed approximately 90% inhibition in the BSA-Glu assay and nearly 100% inhibition in the BSA-MGO assay. Based on these data, IC\({}_{50}\) values were calculated as 10.8 mg/mL and 6.6 mg/mL for aqueous and methanolic extracts, respectively, in the BSA-Glu assay, and 16.6 mg/mL and 9.4 mg/mL, respectively, in the BSA-MGO assay. Numerous studies have highlighted the critical role of AGE in mediating the harmful effects of hyperglycemia, particularly in the development and progression of diabetes-related complications. Among various glycation precursors, MGO has been identified as a major contributor to intracellular AGE accumulation, underscoring its importance as a pathogenic factor in diabetic conditions [23]. One of the primary strategies to inhibit AGE formation involves reducing reactive carbonyl species and scavenging oxidative free radicals. In this context, in vitro studies have demonstrated that trapping MGO can significantly reduce AGE formation, positioning this mechanism as a promising approach to glycation mitigation. In light of this, the MGO-trapping capacity of caro flow extracts was evaluated by testing a range of concentrations (1 to 25 mg/mL) over incubation periods of 24 to 168 h for both aqueous (A) and methanolic (B) extracts (Figure 2). Pyridoxamine was employed as a positive control due to its high reactivity and strong ability to bind and neutralize methylglyoxal effectively [24]. All extracts exhibited MGO-trapping capacity, with higher concentrations resulting in greater MGO-trapping efficacy. In aqueous extracts, 1 mg/mL trapped less than 20% of MGO after 24 h, whereas 25 mg/mL trapped over 60% within the same period, reaching 94% after 168 h. Similarly, the methanolic extract at 1 mg/mL achieved less than 30% trapping, while 25 mg/mL concentration captured 80% of MGO at 48 h and peaked at 97% after one week of incubation. The IC\({}_{50}\) values for MGO-trapping activity are presented in Table 2, expressed both in terms of concentration (mg/mL) and incubation time (h). The methanolic extract exhibited lower IC\({}_{50}\) values than the aqueous extract, indicating higher efficiency in MGO scavenging. Over time, the concentration required to trap 50% of MGO decreased significantly from 16.4 to 8.6 mg/mL for the aqueous extract and from 8.8 to 4.0 mg/mL for the methanolic extract. At fixed concentrations, the methanolic extract reached an IC\({}_{50}\) at 87 h with 5 mg/mL, which decreased significantly to 21 and 15 h at 10 and 25 mg/mL, respectively. Although less effective at lower concentrations (e.g., 10 mg/mL), the aqueous extract achieved a comparable IC\({}_{50}\) to that of the methanolic extract at 25 mg/mL, suggesting a similar trapping capacity at higher doses.