In 1st step we have extracted plant pigments in 100 %acetone .
Step 2 ... Acetone extract is mixed with petroleum ether in separotory funnel .
Step 3 ......Acetone containing chlorophyll is washed with slow addition of water (here we assuming that carotenoid pigment comes in petloum ether layer )
4th step .....For separating xanthophyll from other carotenoid pigments we added ethanol .....Gradient of dark green to pale green is formed from top to bottom ......Now we assuming that lower portion is xanthophyll and upper carotenoid...
Does this procedure is correct and satisfactory for quantification of xanthophyll ?????
Dear sir,
if I understood right, you want to separate xanthophylls from the more lipiphilic carotins. To do so I would recommend an easy separation procedure: First extract your plant material in ethanol or methanol (not aceton, because this will not form a layer with benzine). After extraction add benzine (best: boiling point 40-60°C). Chlorophylls and carotins will migrate into the benzine phase. Xanthophylls will stay in the alcohol.
You can separate and purify the different xanthophylls e.g. by means of a column chromatography.
Hope this will help
Best regards
Peter Richter
FOR SEPARATION AND PURIFICATION USE EITHER PREOPERATIVE TLC (EASY AND FAST ) OR COLUMN CHROMATOGRAPHY
You can follow the Thin layer Chromatography methods for separation and purification of your desire compound from the extract.
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