# Rosemary Leaf Extract Inhibits Glycation, Breast Cancer Proliferation, and Diabetes Risks
## 3 Results and Discussion
### Effect of Rosemary Extract on AGEs, CML and Glycation in Sugar-Protein Rich Foods
Caffeic acid was  
Figure 2: Fluorescence absorbances of AGEs in control and rosemary extract treatments under (**a**) wa 0.75 and (**b**) wa 0.56 conditions (RE1, 1%; RE2, 2%; RE3, 4%). found to reduce approximately 80% of CML at a fortification level of 2.0 g in 100 g bread dough [27]. In the study by [PERSON], the addition of ferulic acid reduced the formation of CML and fluorescent AGEs in vitro by nearly 90% by blocking free amino groups [28]. In addition, ellagic acid and protocatechic acid demonstrated the antiglycative effects on the renal production of CML [29]. Those phenolics have been evidenced to have the potential to trap reactive dicarbonyl species through their benzene rings with one or more hydroxyl groups, thus, forming relatively nontoxic adducts, therefore inhibiting AGEs or CML [30]. Lastly, carnostic acid was proven to reduce AGEs and CML formation via pathways of conjugation with AGEs/CML precursors glyoxal (GO) and methylglyoxal (MGO) [31]. Furthermore, the profiles of glycated \(\alpha\)-lactalbumin (LA), \(\alpha\)-lactoglobulin (LG-A), and \(\beta\)-lactoglobulin (LG-B) in WPI protein were analyzed by LC-MS to investigate the modification of their structure during storage day 1, 3, 5, 7, and 21 under wa 0.75 condition. The molecular weights of LA, LG-A, and LG-B were 14,177, 18,362, and 18,277 Da, respectively (Figure 4a). The longer  
Figure 3: Concentrations of CML formed in control and RE treatment during storage under wa (**a**) 0.75 and (**b**) 0.56 conditions. incubation time induced a higher degree of protein glycation which is indicated by the formation of high molecular weight derivatives of LG-A, LG-B, or LA. As shown in Figure 4b,c, there is no significant difference between the control and RE3 treatment after day 1, since both of them produced high molecular weight derivatives of LG-A, LG-B, and LA with 4 to 8 glucose (G) molecules bound. After day 3, the order of the three highest abundance of glycated LG-A in the control was LG-A + 10G (20,144 Da) \(>\) LG-A + 9G (19,982 Da) \(>\) LG-A + 11G (20,306 Da), while it was LG-A + 9G (19,982 Da) \(>\) LG-A + 10G (20,144 Da) \(>\) LG-A+8G (19,822 Da) in the RE3 treatment (Figure 4d,e). At day 5, higher molecular weight glycated LG-A, LG-A + 13G (20,630 Da) and LG-A + 14G (20,794 Da) were observed in the control rather than RE3 treatment (Figure 4f,g). At day 7, the highest abundance of glycated protein was LG-B + 12G and LG-A + 12G in both the control and RE3 treatment, respectively (Figure 4h,i). However, after 21 days, multiple glycated derivatives occurred in both the control and RE3 treatment (Figure 4j,k). This indicates that the glycation of WPI protein was reduced by the rosemary extract within 21 days under the wa 0.75 condition. In the study of Miroliaei, Melissa officinalis L. extract which also contained in high levels of rosemarinic acid, which demonstrates the protection of BSA protein glycation. This was due to rosemarinic acid's potential to conceal the glycation sites and prevent cross \(\beta\)-structure formation in protein [32]. Besides the activity from multiple phenolic compounds, herb extracts are also confirmed to retard AGE-induced toxicity by the suppression of receptor signaling pathways [32]. Figure 4: Cont. Figure 4: Mass spectrums of protein glycation in control and RE treatment during storage.