Analysis of chlorophyll and carotenoid compounds in eucalyptus leaves

Abstract This protocol aims to extract and measure the amount of chlorophyll and carotenoids in leaf material using spectrophotometry. An ethanol extraction is used to analyse both chlorophyll a and b and carotenoids in leaf material with compounds quantified using absorbance across a range of wavelengths. This protocol uses ethanol as the extracting solution due to its affordability and safety compared to other reagents. We report a methodology that extracts pigments from whole leaf discs. The combination of extraction from leaf discs and ethanol extraction ensures this protocol is safe and rapid to complete. The protocol includes three phases: a preparation phase where samples are punched and added to ethanol (approximately 3 hours for 100 samples), an extraction phase where samples are left overnight, and a measurement phase where absorbance at a range of wavelengths is recorded using spectrophotometry (approximately 1 hour for 100 samples).