## 2 Materials and Methods
### Fatty Acid Composition and Nutritional Indices  
Minced meat products and flour samples were thawed overnight at 4 \({}^{\circ}\)C. The following day, fatty acids were extracted and methylated according to the method described by [PERSON] et al. [32]. Briefly, the samples were placed in a screw-capped Pyrex tube, and aqueous KOH solution and methanol were added. The tubes were placed in a water bath (55 \({}^{\circ}\)C; 90 min) and vigorously shaken by hand for 5 s every 20 min to properly permeate, dissolve, and hydrolyse the samples. Tubes were cooled in iced water bath, and aqueous H\({}_{2}\)SO\({}_{4}\) was added. The contents of the tubes were gently mixed by inversion, and then the tubes were placed again in the water bath (55 \({}^{\circ}\)C; 90 min) and shaken for 5 s every 20 min. Tubes were again cooled in iced water bath. Hexane was added, and the tubes were vortex-mixed for 5 min; following that, the tubes were centrifuged at 1100\(\times\)\(g\) for 10 min. The upper phase was filtered through a 0.45 \(\upmu\)m pore-size PVDF syringe filter, transferred into amber GC vials, and stored at \(-\)20 \({}^{\circ}\)C until analysed.  
Fatty acid methyl ester analysis was performed on an Agilent Technologies 6890N GC (Agilent Technologies, Inc., Santa Clara, CA, USA) equipped with a flame ionisation detector (FID) and an Agilent 7683 autosampler. Chromatographic separation was achieved with a fused silica capillary column (Column DB--23 0.25 mm film ID \(\times\) 60 m \(\times\) 0.25 mm; Agilent Model 122-2362). The injector temperature was set to 250 \({}^{\circ}\)C. The GC conditions were as follows: carrier gas, He; split mode injection, 50:1 (3 \(\upmu\)L); injector and FID temperatures, 250 \({}^{\circ}\)C and 300 \({}^{\circ}\)C, respectively; and initial oven temperature, 110 \({}^{\circ}\)C for 6 min, increased at 11 \({}^{\circ}\)C per min to 165 \({}^{\circ}\)C, increased at 15 \({}^{\circ}\)C per min to 195 \({}^{\circ}\)C, increased at 7 \({}^{\circ}\)C per min to 230 \({}^{\circ}\)C, and held at 230 \({}^{\circ}\)C for 7 min. Fatty acids were characterised using three commercial standard mixtures: (a) a 37-component FAME mix (Supelco, Bellefonte, PA, USA, 47885-U), (b) PUFA-2 from animal source (Supelco, 47015-U), and (c) a blend of cis- and trans-9,11- and -10,12-octadecanenoic acid methyl esters (Sigma, St. Louis, MO, USA, O5632-250 MG) as reference standards. Quantification of fatty acids was performed through peak area measurement, and the outcomes are presented as a percentage (%) of the total peak areas for all quantified acids. The fatty acids were categorised into saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), and unsaturated fatty acids (UFA).  
The fatty acid profiles were used to determine the nutritional indices associated with healthy fat intake. The applied indices are those outlined in the recent review conducted by [PERSON] and [PERSON] [33] for meat products. These researchers systematically compiled information on fatty acid profiles published since 2000 to enhance the understanding of the implications and applications of diverse nutritional indices.  
Polyunsaturated fatty acids/Saturated fatty acids ratio  
\[\text{PUFA/SFA}=\frac{\Sigma\text{PUFA}}{\Sigma\text{SFA}}\]  
Atherogenicity Index  
\[\text{AI}=\frac{(\text{C12:0}+(4\times\text{C14:0})+\text{C16:0})}{\Sigma\text{ UFA}}\]  
Thrombogenicity Index  
\[\text{TI}=\frac{\text{C14:0}+\text{C16:0}+\text{C18:0}}{\left((0.5\times\Sigma \text{MUFA})+(0.5\times\Sigma\text{n}-6\text{PUFA})+(3\times\Sigma\text{n}-3 \text{PUFA})+(\text{n}-3\text{PUFA}/\text{n}-6\text{PUFA})\right)}\]  
Hypocholesterolemolaemic/hypercholesterolaemic fatty acid ratio (h/H)  
\[\text{h}/\text{H}=\frac{\text{C18:In}-9\text{cis}+\Sigma\text{PUFA}}{(\text{C1 2:0}+\text{C14:0}+\text{C16:0})}\]