## 2 Materials and Methods
### Analysis of Fatty Acid Profile  
Fatty acid profile was determined using the method described by [PERSON] [20] with modifications. Briefly, 40 mg of lipid extract was transferred into a Pyrex tube. One milliliter of sodium monoxide in methanol (0.5 M) and 1 mL of 0.6 mg/mL internal standard, C13:1 in n-heptane, were added to the Pyrex tube containing the lipid extract and incubated at 50 \({}^{\circ}\)C for 15 min. The mixture was cooled at room temperature for 5 min. To this mixture, 1.5 mL of 5% HCl in methanol was added then incubated at 80 \({}^{\circ}\)C for 15 min, followed by cooling at room temperature for 7 min. To the cooled mixture, 2.5 mL of 6% K\({}_{2}\)CO\({}_{3}\) and 1 mL of n-heptane were added, vortex-mixed for 30 s and centrifuged at 4000\(\times\)\(g\) at 4 \({}^{\circ}\)C for 5 min. The upper phase containing fatty acid methyl ester was injected into a gas chromatograph (Agilent 7890 gas chromatograph) equipped with an Agilent CFT modulator for GCxGC analysis, flame-ionization detector and Agilent 7693 autosampler. The gas chromatographic conditions were as follows: initial oven temperature 45 \({}^{\circ}\)C, isotherm (hold time) 2.0 min, heating rate 50 \({}^{\circ}\)C/min up to 170 \({}^{\circ}\)C, isotherm (hold time) 25 min, temperature increase by 2 \({}^{\circ}\)C/min up to 240 \({}^{\circ}\)C, isotherm (hold time) 16 min, injection port temperature 270 \({}^{\circ}\)C and detector port temperature 250 \({}^{\circ}\)C. The injected volume was 1 \(\upmu\)L in a split mode (160:1). Hydrogen was used as carrier gas. As the primary column, a Supelco SP-2560 (75 m \(\times\) 0.18 mm \(\times\) 0.14 \(\upmu\)m film thickness) with an initial flow rate of 0.25 mL/min, which was increased to 0.4 mL/min at a rate of 0.002 mL/min, was used and as the secondary column, an Agilent J&W HP-5 ms (3.8 m \(\times\) 0.25 mm \(\times\) 0.25 \(\upmu\)m film thickness) with an initial flow rate of 22 mL/min, increased to 35 mL/min at a rate of 0.18 mL/min, was adopted. The valves were set to a modulation delay of 1 min, a modulation period of 2.9 s and a sampling time of 2.77 s. One microliter sample was injected in the pulsed split mode at a pressure of 25 psi for 0.3 min and a split ratio of 160:1. The split-splitness inlet was run at a temperature of 270 \({}^{\circ}\)C. The resulting two-dimensional chromatograms were analyzed with comprehensive GCxGC software (GC Image R 2.2  
\begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Name of Varieties** & **Year of Release** & **Source** & **Seed Color** & **Local Name** & **Adaptation Zone (Altitude)** \\  & & & & **(masl)** \\ \hline DZ-01-99 & 1970 & Debrezeit & Brown & Asgori & 1600–2400 \\ \hline DZ-01-2423 & 2005 & Adet & Brown & Dima & 2000–2600 \\ \hline DZ-01-2053 & 1999/8 & Holetta & Brown & Holeta Key & 1900–2700 \\ \hline DZ-01-1278 & 2000 & Holetta & White & Ambo Toke & 2200–2400 \\ \hline DZ-01-1681 & 2002 & Debrezeit & Brown & Key Tena & 1600–1900 \\ \hline DZ-01-2675 & 2005 & Debrezeit & White & Dega Tef & 1800–2500 \\ \hline DZ-Cr-387 & 2006 & Debrezeit & Very white & Quncho & 1800–2400 \\ \hline White amaranth & - & Bench Majji & White & Katila & - \\ \hline Brown amaranth & - & Bench Majji & Brown & Katila & - \\ \hline Red amaranth & **-** & Bench Majji & Pale red & Katila & - \\ \hline \hline \end{tabular}
\end{table}
Table 1: Description of teff and amaranth varieties used in the study [17; 18].  
GCXGC: Zoex Corp., Houston, TX, USA). The fatty acid composition was expressed using the formula described below.  
\[\text{Total fatty acids }(\text{mg/g of sample})=\frac{(V_{TP}-V_{IS})*\text{mg of internal standard}}{(V_{IS}*\text{sample weight }(\text{g}))} \tag{1}\]  
where \(V_{TP}\) is the volume of total peaks and \(V_{IS}\) is the volume of internal standard.