Then, 43 mL of water was added and the solution was mixed for 1 h. The supernatant was separated through centrifugation and acidified to pH 5 to isolate 2.36 g EFA (19 wt. % of DOB). The precipitate was acidified to pH 4.7 to obtain 0.37 g of 22-hydroxy docosanoic acid (3 wt. % of DOB). In both cases, centrifugation was used to recover the fatty acid as the precipitate. The 22-hydroxy docosanoic acid was not further used in this study. Both suberin fatty acids were freeze-dried and stored at room temperature.