Eleanor Gibbs

An evaluation of reversed phase partition for thin-layer chromatographic identification of chlorophylls and derivatives☆

Abstract Separation and identification of chlorophylides a and b, pheophorbides a and b, as well as chlorophylls a and b and pheophytins a and b by reversed-phase partition thin-layer chromatography, was shown to be effective. Pigment quantities in the order of 0.5 nmole could be identified. Criteria of evaluation were location, color and stability of pigment zone as viewed under white light and ultraviolet light. Investigations in which oil-impregnated thin-layers of powdered sugar, Kieselguhr G and a mixture of Kieselguhr G and Silica Gel G were used were reported. The formation of artifacts occurred when chromatographing purified pigments on inorganic thin-layers. The location of artifact formation is considered to be on the portion of the plate coating which was not oil-impregnated, Addition of an antioxidant such as butylated hydroxytoluene to pigment solutions prior to application on inorganic layers was shown to prevent artifact formation. Identification characteristics for allomerized chlorophylls a and b; methyl chlorophyllides a and b; and methyl pheophorbides a and b on oil-impregnated plates were indicated.