2.4. Analysis of acrylamide
Ground samples (5 g) were dissolved in 50 mL hot water and filtered using 0.45 mm Whatman filter paper.
Solid Phase Extraction (SPE) C18 column was conditioned using 3 mL acetone and 3 mL formic acid.
Filtered sample was applied to SPE tube at which the sample solution is allowed to pass through tube with
gravity flow. SPE tube was washed using 2 mL distilled water and vacuum was used for 2 minutes to dry the
excess water. Sample was eluted using 3 mL acetone. The eluted sample was filtered using 0.45 μL syringe
filter and kept at -20o
C until further GC-FID analysis. Analysis of acrylamide was carried out using Gas
Chromatography (Shimadzu 2010) equipped with Flame Ionization Detector with a temperature 260°C. RTX-
5 column (30m x 0.25mm I.D. x 0.25 μm film thickness) was used with injector temperature of 260°C, helium
gas at constant pressure as carrier gas, and with oven temperature of 100 °C (hold 0.5 min) to 200°C at
15°C/minute [1]