Quantitative Measurement of Leaf Chlorophylls by Spectrophotometry of Their Pheophytins in Aqueous Alcoholic Extracts

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Abstract
Methods are described for analysis of leaf chlorophylls by spectrophotometry and colorimetry of their pheophytins in acidified 80% aqueous ethanolic extracts. Of 2 methods of sample preparation, that involving freezing in liquid N and extraction by simmering 80% aqueous ethanol was found to yield results with less variability than the method involving extraction of a dried sample by grinding with hot 80% ethanol. With a given method of sample preparation, the 2 spectrophotometric and the colorimetric methods of assay yielded results of practically equal precision, but the Cary recording spectrophotometer employed was the superior instrument with respect to sensitivity and precision of results. The data indicate that such analytical procedures can be employed to measure differences of 1% or greater in the chlorophyll content in leaves.

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