Abstract
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, SpainMaterial 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.
Highlights
Laboratory analysis of pigments in tree leaves has traditionally been conducted by extraction methods using several water-soluble organic solvents followed by spectrometric determination based on the maxima absorption spectra of Chlorophylls (Chls) and carotenoids in the red and blue spectral regions (Minocha et al, 2009)
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
We investigated the drawbacks in the use of DMSO as solvent for pigment quantification in Quercus ilex L. leaves used for SPAD calibrations
Summary
Laboratory analysis of pigments in tree leaves has traditionally been conducted by extraction methods using several water-soluble organic solvents followed by spectrometric determination based on the maxima absorption spectra of Chlorophylls (Chls) and carotenoids in the red and blue spectral regions (Minocha et al, 2009). 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. 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.
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