Quantification of Egg Yolk Fatty Acids

Capillary high-resolution gas chromatography (HR-GC) with a CP-Sil 88 CB column was used to analyze the fatty acids in all sample extracts. The GC column was installed on a PerkinElmer gas chromatograph CLARUS 680 equipped with a flame ionization detector and split injection, as previously described [53 (link)]. In brief, hydrogen was employed as the carrier gas at a flow rate of 1 mL × min⁻¹ while the split ratio was 1:20, with the injector and detector adjusted at 260 °C and 280 °C, respectively. The GC oven temperature programme was 150 °C for 5 min; 2 °C/min heating until 200 °C and held for 10 min; and 1 °C/min heating until 225 °C and held for 20 min. To calibrate the reference standard mixture “Sigma FAME”, the methyl esters of C18:1cis-11, C22:5n-3, and C18:2cis-9, and trans-11, C22:4n-6, and C18:4n-3 were employed. After GC analysis of 5 samples, the 5-point calibration of single fatty acids assessed were ranged between 16 and 415 µg/mL. Fatty acid concentrations were displayed in milligrams per gram of egg yolk.