## 2 Materials and Methods
### Determination of Color Characteristics  
The samples' color properties were analyzed with a Konica Minolta CM-2300d spectrophotometer, following the methodology described in the study by [PERSON] et al. (2019), which evaluated antioxidant potential in minced beef meat. Before the measurements, the spectrophotometer underwent both zero and white calibration according to the manufacturer's guidelines. The final result for each sample was derived as the arithmetic mean of the five measurements.  
Analysis of Molecular Weight Distribution of Protein Fractions in Samples Carried Out Using One-Dimensional Electrophoresis  
A 100 mg sample was mixed with 500 \(\upmu\)L of a lysis buffer (4.5 M urea, 2.5% \(\beta\)-mercaptoethanolhanol, 1% Triton X-100, and 1% ampholytes, with pH 3-10) and centrifuged at 14,000 rpm for 20 min. The supernatant was carefully separated, and an equal volume of protein buffer was added to the mixture. The protein buffer was prepared by combining 1 mL of 10% sodium dodecyl sulfate (SDS), 250 \(\upmu\)L of concentrated \(\beta\)-mercaptoethanolhanol, 625 \(\upmu\)L of 0.5 M Tris-HCl, 1.5 g of urea, and bromophenol blue as a dark stain. The total volume of the solution was adjusted to 5 mL with distilled water. Following this, the samples were heated in a boiling water bath for 5 min. Protein visualization was achieved using Coomassieie G-250 staining. The staining solution consisted of 10% acetic acid, 25% isopropanol, and 0.05% Coomassieie G-250. To eliminate excess unbound dye, 10% acetic acid was used as a washing solution. One-dimensional electrophoresis gels in their wet state were utilized for densitometric analysis. High-resolution digital images were captured using a Bio-5000 Plus scanner (Serva, Heidelberg, Germany) at 600 ppi in 2D-RGB mode. The obtained digital images were subsequently refined using graphic editing software for further analysis. The version number of ImageJ was 1.53t, and it was accessed via the official website ([[https://imagej.net/ij/](https://imagej.net/ij/)]([https://imagej.net/ij/](https://imagej.net/ij/))) on 20 August 2024.  
Figure 1: The sausage with purslane (**a**) and without purslane (**b**).  
Determination of Ferric-Reducing Antioxidant Power (FRAP) and Antioxidant Activity Using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Assay  
The ferric-reducing antioxidant power (FRAP) assay was conducted using BHT and \(\alpha\)-tocopherol as standard antioxidants. To perform the assay, 1 mL of the extract at varying dilutions was combined with 2.5 mL of phosphate buffer (0.1 M; pH 6.6) and 2.5 mL of potassium ferricanide solution (1%, \(w/v\)). The mixture was incubated at 50 \({}^{\circ}\)C for 20 min. Following incubation, 2.5 mL of trichloroacetic acid solution (10%, \(w/v\)) was added. From this mixture, 2.5 mL was withdrawn and combined with 2.5 mL of deionized water and 0.5 mL of ferric chloride solution (0.1%, \(w/v\)). The solution was allowed to stand for 30 min before the absorbance was measured at 700 nm. The FRAP values were expressed as milligrams of gallic acid equivalents (GAEs) per gram of dry extract (mg GAE/g).  
For the DPPH radical-scavenging activity assay analysis, 2 mL of a DPPH solution (0.1 mg/mL in methanol) was mixed with 2 mL of the sample solutions at a concentration of 200 ug/mL. All analyses were performed in triplicate. The reaction mixture was shaken and incubated in the dark at room temperature for 30 min. After incubation, the absorbance was measured at 517 nm against a blank. Ascorbic acid was prepared as a positive control in a similar manner, with the antioxidant solution being replaced accordingly.