Lutein extraction after exposure

Samples were processed immediately on completion of exposure to different temperatures and pHs for specified time. Lutein was extracted from samples Bhat et al. (2021a) using dichloromethane:methanol:hexane in the ratio 2:2:4:3 (v/v/v/v). The mixture was vortexed (1 min), centrifuged (3000 rpm, 5 min) and upper layer of hexane was collected. Repeated extraction was performed with the lower aqueous layer using dichloromethane:hexane (2:3 v/v) and centrifugation (1000 ×g, 5 min) at room temperature. The upper hexane layer was collected, pooled and dried over a stream of nitrogen. The samples were redissolved in a known amount of mobile phase and analyzed by HPLC.