## 2 Results and Discussion
### Effects of Germination on the FA Profile of IWG  
FAs play a crucial role in perennial grains by contributing to several key aspects of plant health, plant-plant, plant-microbe, and plant-environment interactions, nutrition aspects, and agricultural sustainability. They also play an important role in consumer acceptability as they serve as precursors to the formation of flavor compounds, such as aldehydes, that have a lower threshold [23]. The fatty acid profile of samples involves the extraction of lipids, derivatization of fatty acids, and chromatographic analysis of fatty acid methyl esters (FAMEs). For lipid extraction, solvent extraction methods such as the Folch or Bligh and Dyer methods are commonly used to isolate lipids from samples. These methods involve solvents such as chloroform and methanol to effectively extract fats and oils. To analyze FAs, they are often converted into fatty acid methyl esters (FAMEs) through transesterification or direct methylation. This makes the FA volatile and suitable for analysis using gas chromatography (GC). The primary technique for analyzing FAMEs is GC. FAs are separated and identified based on their retention times on a capillary column. The detection of the compounds is accomplished by a flame ionization detector (FID) (widely used in GC for detecting fatty acids) or mass spectrometry (MS) [24]. The overall FA profile of IWG samples exhibited a high concentration of linoleic acid (C18:2n6, \(\sim\)62%), an essential polyunsaturated fatty acid (PUFA), followed by oleic acid (C18:1, \(\sim\)17%) and palmitic acid (C16:0, \(\sim\)12%) (Table S1--Supplementary Material). The total FA concentration varied from 16.41 to 21.78 mg/g of sample. This profile is similar to those found in wheat germ and sunflower oil, which present more than 60% of C18:2n6 in their composition. However, the content of C18:1 is higher in sunflower oil (28%) and lower in wheat germ (13%) when compared to IWG [25; 26]. A significant reduction (from 13.76 mg/g on the control to approximately 10 mg/g after 6 days of germination) in the concentration of linoleic acid (C18:2n6) was observed in all temperatures evaluated (Table 1). Similarly, a reduction in the concentration of oleic acid (C18:1) and palmitic acid (C16:0) was observed within the germination process. However, the overall FA profile was not affected (Table S1). Germination time had a more significant effect on the FA content than the temperature (Table S1). Figure 1: Diagram of the IWG germination process. The germination process activates hydrolytic enzymes, which promote an increase in oligosaccharides, monosaccharides, oligosaccharides, oligopeptides, free amino acids, and free FAs by mobilizing them from the aleuorne layer and scutellum to the starchy endosperm [9]. Therefore, a reduction in the FA content was expected during germination. Moreover, the depletion of FAs can be associated with their use as an energy source for protein synthesis in the growing vegetable [27]. Lipids are abundant in cereal tissues and can be used to provide energy during germination. The mobilization of triacylglycerglycerol begins with germination, leading to the conversion of oil to sugars through the \(\beta\)-oxidation and glyoxylate cycles [28]. The process begins with the liberation of esterified FAs from triacylglycerglycerols via lipase. Lipases are ubiquitous, being widely distributed in plants, animals (including insects), and microorganisms. They exhibit high specificity and selectivity for their substrates. In plants, the growing embryo has been reported to have the maximum lipase activity [29]. [PERSON] and co-workers [4] reported an increase in lipase activity in IWG. The authors found that within three months of storage, lipase activity significantly depleted triacylglycer levels, leading to a peak in free FA content. These free FAs can serve as an energy source through \(\beta\)-oxidation and glyoxylate cycles during germination, ensuring the conversion of FAs into carbohydrates essential for seedling growth [30].