Elsevier

Methods in Enzymology

Volume 23, 1971, Pages 586-602
Methods in Enzymology

[56] Quantitative determination of carotenoids in photosynthetic tissues

https://doi.org/10.1016/S0076-6879(71)23132-3Get rights and content

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This chapter discusses the quantitative determination of carotenoids in photosynthetic tissues. A useful separation of carotenoid pigments into nonpolar carotenes and carotenoid esters in one group, and polar xanthophylls (containing hydroxyl groups) in the other may be accomplished by partitioning the pigments between petroleum ether and aqueous methanol. This can be done by dissolving the extract in aqueous methanol in a separatory funnel and adding an equal amount of petroleum ether. In cases where the carotenoid components are stable toward alkali, it is advantageous to include saponification in the isolation procedure for removing chlorophyll and other saponifiable matter. The pigment is taken to dryness (oil pump); all acetone must be removed from the extract to avoid base-catalyzed formation of condensation products of acetone. The residue is dissolved in a small volume of ether, and the same volume of 10% methanolic KOH solution is added. The approximate content of total carotenoids in an extract may be determined by measuring the optical density of a sample suitably diluted.

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