Short communication: Aqueous-acetone extraction improves the drawbacks of using dimethylsulfoxide as solvent for photometric pigment quantification in Quercus ilex leaves

Rosario Gonzalez-Cascon, Leticia Jiménez-Fenoy, Irene Verdú-Fillola, M. Pilar Martín


Aim of study: We evaluated the use of dimethylsulfoxide (DMSO) for the photometric determination of chlorophyll (Chla and Chlb) and carotenoids in Quercus ilex L. leaves by comparative analysis using aqueous-acetone extraction.

Area of study: a Q. ilex dehesa in Las Majadas del Tietar, Cáceres, Spain

Material and methods: Q. ilex leaves were sampled during two vegetative periods. Field SPAD-502 Chlorophyll measurements and photometric chlorophyll determinations were performed.  Two procedures were used: extraction of intact foliar discs at 65ºC with DMSO and fine foliar powder with cold aqueous-acetone.

Main results: DMSO produced Chlb overestimation and different fitting performance for SPAD vs pigment calibrations (R2=0.64, RMSE=0.20, p<0.0001 for Chla (µg/cm2); R2=0.33, RMSE=0.23, p<0.0001 for Chlb (µg/cm2) and R2=0.50, RMSE=0.23, p<0.0001 for carotenoids (µg/cm2)). Aqueous-acetone provided more accurate predictions (R2=0.90, RMSE=0.16, p<0.0001 for Chla and R2=0.91, RMSE=0.16, p<0.0001 for Chlb, R2=0.90, RMSE=0.02, p<0.0001 for carotenoids) and mean ratio Chla/Chlb=3.6 inside the range for sun exposed leaves.

Research highlights: Oxidizing conditions and polyphenol concentrations in Q. ilex leaves generated brown colorations in the DMSO extraction procedure, interfering with the photometric measurements in the red-orange region. Aqueous-acetone extraction was free from interference. DMSO should be avoided for pigment determination in Q. ilex leaves or when comparing different tree species.


chlorophyll; carotenoids; DMSO; SPAD-502; phenolic compounds

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DOI: 10.5424/fs/2017262-11099