Separation, quantification, spectral properties and stability of photosynthetic pigments on CN-coated HPTLC plates

Summary

Chlorophylls, phaeophytins a and b, β-carotene, lutein, violaxanthin and neoxanthin can be separated in 15 min on HPTLC, CN-coated sheets using chloroform-hexane-methanol (25-70-05 v/v). The algebraic characteristics of calibration curves and of the absorbance decay with time have been determined for each component. Solid phase spectra have been established from 370 to 700 nm and their variations have been examined with respect to the amounts of pigments spotted on the plates and to the storage time of chromatograms in the dark at 4°C.

Pigments extracted with chloroform from barley leaves were analysed using the described method. A 5% accuracy is normally to be expected, and the sensitivity ranges from 0.5 pmol (β-carotene) or 1 pmol (chlorophylls a and b) to 12 pmols (lutein) in quantitative determinations at 425 nm.