2.5.1.3. Carotenoids separation. About 20 ml distilled petroleum ether was placed in a 250 ml separating funnel wrapped with black cloth. The coloured acetone was carefully added to the pet- roleum ether through a funnel, gently flowing down the wall of separating funnel. The vacuum flask was rinsed with 1-2 ml ace- tone. Then, 150 ml distilled water was added to the separating fun- nel (flowing down the funnel wall), the mixture was left undisturbed for 5-10 min to allow for separation into organic and aqueous layers. The aqueous layer, containing acetone, was discarded. This step was repeated for 3-4 times with 100 ml dis- water added each time until the residual acetone was removed. The petroleum ether extract in the separating funnel was collected, through a funnel containing small amount of anhy- drous sodium sulfate on a filter paper, into a 25 ml volumetric flask covered with aluminum foil The separating funnel was rinsed with about 2 ml petroleum ether using a pipette, The eluent volume was made up to 25 ml with petroleum ether. The flask was capped and gently mixed. For determination of the carotenoids, absorbance of the orange-coloured eluent was measured at 450 nm in a spec trophotometer (UV-1601, Uv-visible, Shimadzu, Tokyo, Japan) l the preparative and extraction procedures were performed in dim light and/or excluded light as described.