Green Improvement of Thin Layer Chromatography for Separation of Spinach Pigments
Thin layer chromatography(TLC)is an important experimental technique for rapid separation and qualitative analysis of a small amount of substances.In the original experiment of separating spinach pigment by TLC,the selected development agent uses a large amount of benzene,a carcinogen,and there usually occur spot tailing and unclear separation of pigment points during the experiment.In order to optimize the experimental effect,avoiding use of carcinogenic benzene,the non-polar solvent petroleum ether,n-hexane,cyclohexane and polar solvent ethanol,ethyl acetate and acetone have been combined for multiple times to separate spinach pigment by thin layer chromatography.The results show that when the volume ratio of n-hexane-acetone is 7:3,the separation effect is the best.When the Rf values of carotene,pheophytin,chlorophyll a,chlorophyll b and lutein are 0.92,0.85,0.42,0.28 and 0.72 respectively,and the pigment spots are complete and clarity.This method has been verified,and the result shows good reproducibility and durability.
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