Comparison of solvent regimes for the extraction of photosynthetic pigments from leaves of higher plants

The relative efficiency of methanol- and acetone-based solvents for the extraction of pigments from photosynthetic tissues of plant was compared, together with the advantages of multiple versus single extractions. The two commonly employed triple acetone extractions (100 : 80 : 80% and 85 : 100 : 100%) performed comparably for most pigments and for all plant species tested. Single extractions with either 96% methanol or 85% acetone failed to extract the more hydrophobic pigments, especially β-carotene. We conclude that multiple extractions that combine pure and aqueous (80-85%) acetone are preferable for extraction of the full range of pigments. These results suggest that previous studies that have utilised aqueous methanol (especially in a single extraction) have probably underestimated the concentration of β-carotene relative to other pigments.