Green vegetables carotenoids, extracts/extraction

Khachik, R, Beecher, G.R., and Whittaker, N.R, Separation, identification and quantification of the major carotenoid and chlorophyll constituents in extracts of several green vegetables by liquid chromatography, J. Agric. Food Chem., 34, 603, 1986. 

A general procedure that our laboratory generally employs is the addition of an equal amount of methanolic 10% potassium hydroxyde (KOH) to an ethereal carotenoid extract. This solution is bubbled with N2 and allowed to stand overnight at room temperature. Other conditions that shorten time at room temperature have also been used, such as saponification of the dichloromethane (CH2CI2) extract with the same amount of 10% KOH in MeOH for 1 hr (peppers and fruits ) and ethereal extract treated with 30% methanolic KOH under N2 for 3 hr (green leaves, vegetables and fruits ). 

Carotenoids are 40-carbon compounds made up of eight isoprene units. Carotenoids serve as light absorbing molecules in photosynthesis and also protect certain tissues from the deleterious effects of light. Beta-carotene is the precursor of vitamin A (which does not occur in plants). Beta-carotene occurs abundantly in the liver oils of fish and is extracted from that matrix commercially. Nutritionists recommend tomatoes, carrots, and green vegetables in daily quantities in order that carotenoids can be converted into vitamin A, a compound necessary for vision and other vital functions. 

Ishida and Chapman " studied ethyl lactate as a green solvent to extract carotenoids from several vegetable sources. The authors determined the carotenoid yield (pg of carotenoid per g of dry weight) at different conditions and the results were compared with extraction carried out with ethanol (another green solvent), ethyl lactate/ethanol mixtures, as well as methylene chloride/methanol/H20 (40/40/20) and ethyl acetate, being the most commonly used solvent for extracting carotenoids for utilization in food products, but not considered to be an environmentally friendly solvent. 

Roy et al. (1991) extracted the pigments of green leafy vegetables in methanol-acetone-diethyl ether (1 1 1) the extracts were evaporated, reconstituted in the same solvent, and separated by TLC on silica gel G plates with a chloroform-petroleum ether-toluene-acetone (5 4 4 3) mobile phase. Pheophorbide b, mixed carotenoid, pheophorbide a, chlorophyll b, pheophytin b, chlorophyll a, pheophytin a, and carotene bands were obtained. This method can be used to detect artificial and natural colorants in food products. 

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