The petroleum ether extract in the separating funnel was collected, through a funnel containing small amount of anhydrous 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