*2.2. Physico-Chemical Analysis*

The moisture, protein, fat, ash, and fiber content of the raw materials as well as of the RTC-GF snack were determined as per the method prescribed by AOAC [20]. Total carbohydrates were estimated by the difference method, as in Yadav et al. [21]. The energy content in kcal/100 g was calculated as per Sehgal et al. [11]. For mineral estimation, one gram of sample was digested in a combination of nitric acid and perchloric acid (3:1), made up to a volume of 50 mL using deionized water. The minerals present in it were then estimated by plasma atomic emission spectrometry (Thermo Scientific, Waltham, MA, USA), as shown by Kaur et al. [22], where the samples were digested using a di-acid mixture, made up to the required volume with deionized water, filtered, and then further used for estimation. The amino acid analysis was carried out according to the method given by Musaalbakri et al. [23]. The estimation was done using HPLC, undertaking the hydrolysis of the protein with 6 M HCL containing 0.1 per cent phenol 110 ◦C for 24 h. Methionine was determined after pre-hydrolysis with performic acid oxidation before HPLC analysis, whereas tryptophan was determined by alkaline hydrolysis. Amino acids were expressed as g/100 g.

**Figure 1.** Process for Preparation of RTC-GF Snack.

The samples were analyzed for fatty acid composition by gas liquid chromatograph (model 7820A series, Agilent Technologies, Palo Alto, CA, USA) equipped with a flame ionization detector with a CP-Sil 88 (25 m × 0.25 mm × 0.20 mm) FAME column. The temperature of the oven, detector, and injector were maintained at 210, 240, and 230 ◦C, respectively. Two μL of the sample were injected at a split rate of 10:1. Individual fatty acids were expressed as a percentage of total fatty acids, according to Hassanien et al. [24].

The color of the RTC-GF was determined in terms of L\*, a\*, and b\* values where L\* measured lightness ranging from black (L = 0) to white (L = 100), a\* measured red (+) or green (-), and b\* measured yellow (-) or blue (-) using a Minolta spectrophotometer colorimeter (ModelCM-508d). Hue angle (h◦) and chroma (C) were measured as per Singh et al. [25].
