The use of acetone and methanol in the estimation of chlorophyll in the presence of phaeophytin

(1)Where grinding or maceration of plant tissue is impractical, immersion in methanol should be used. (2)The adaption to aqueous methanol of existing techniqus for distinguishing chlorophyll and phaeophytin in aqueous acetone was studied in detail. In methanol the spectra of both phaeophytin a and b were found to be pH sensitive. A method developed involving changes in absorbance at 665 nm is less precise than similar methods using acetone. (3)The spectra of phaeophytin b in methanol at different pH levels are anomalous. Changes in absorbance between 440 and 410 nm cannot be used for the estimation of phaeophytin. (4)Plant pigments can be transferred from methanol to aqueous acetone without degradation of chlorophyll. Modified standard techniques may then be used to measure chlorophyll a and phaeophytin a content.